CASE CLOSED … what really happened in the 2001 anthrax attacks?

* Ivins’ lab notebook pages show what Dr. Ivins was scheduled to do on 8/14, 8/21, 8/23, 9/4, 9/6, 9/18, 9/20, and 10/2/2001 … the notebook page below says the research was postponed indefinitely on October 3, 2001 … what corroborating documents exist regarding this series of experiments?

Posted by Lew Weinstein on June 3, 2011

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11 Responses to “* Ivins’ lab notebook pages show what Dr. Ivins was scheduled to do on 8/14, 8/21, 8/23, 9/4, 9/6, 9/18, 9/20, and 10/2/2001 … the notebook page below says the research was postponed indefinitely on October 3, 2001 … what corroborating documents exist regarding this series of experiments?”

  1. Lew Weinstein said

    We need the full record of what Ivins did in the lab on the nights in question …

    and as much corroborating documentation as exists to prove he did the experiments he said he did. Such as … inventory records of materials used … withdrawal of animals from cages … use of other facilities such as those for infection by airborne pathogens … any correspondence with others regarding the ongoing experiments … any statements by others that they saw Ivins doing these experiments or talked with him about them … any reports after the experiments as to the results obtained.

    If all that exists, then the FBI assertion that Ivins was making powdered anthrax on those nights is a crock … and they knew it!

    If, on the other hand, there is nothing to corroborate what Ivins said he did, then there are some serious questions left hanging.

    • DXer said

      You raise a fascinating point given the amount that the FBI redacted in materials previously provided about the postponment on October 3, 2001. While I go look for that document, let me provide some emails from August 2001 to see what light, if any, they shed.

      From: To: Subject: Date: Attachments:
      Ivins, Bruce E Dr USAMRIID
      Objectives Monday, July 23, 2001 9:49:40 AM
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      Here are objectives for me for the coming year. – Bruce
      From: To: Subject: Date:
      YESSSSS!!!
      Ivins, Bruce E Dr USAMRIID
      FW: Animal Proposal 01-28 Monday, July 23, 2001 2:57:05 PM
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      >—–Original Message—– >From: >Sent: Monday, July 23, 2001 2:55 PM >To: Ivins, Bruce E Dr USAMRIID >Subject: Animal Proposal 01-28 > > >Bruce – Approved and assigned protocol number B01-11. >
      >
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      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      RE: follow up on MTA Tuesday, July 24, 2001 11:15:01 AM
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      Sure, It’s
      Thanks – Bruce Ivins
      >—–Original Message—– >From: >Sent: Tuesday, July 24, 2001 10:29 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: follow up on MTA > >Hi, >I’m replacement. forwarded the MTA info with to me and I’m in the process of completing it. Since calling him has been unsuccessful, could you please provide me with his e-mail address. > >Thanks.
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      From: To: Cc: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      RE: Dugway Wednesday, July 25, 2001 9:27:57 AM
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      Thanks, I’m on vacation for a week, until August 1. If you have information, please forward it to
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      – Bruce
      and as well. Talk to you later!
      —–Original Message—– From: Sent: Wednesday, July 25, 2001 9:35 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Dugway
      Hi Bruce, I’m just getting back today from being gone for three weeks. I’ll
      track it down as I catch up today and tomorrow and let you know where we stand. Good to hear from you.
      (6)
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Tuesday, July 17, 2001 6:01 AM To: Subject: FW: Dugway
      (6) How are things going on your end with respect to the contract? According to our contract person, (below) it was sent and should be there. I’m looking forward to working with you again on these. – Bruce
      > —–Original Message—– > From: > Sent: Tuesday, July 10, 2001 10:16 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: Dugway > > Bruce, > > Money and statement of work went to Dugway on Monday. They confirmed receipt. > > > > (6) >
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      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      RE: MTA Wednesday, August 01, 2001 10:29:29 AM
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      – (T6h)is sounds good. Or, perhaps they could send us in detail what they intend to do(?) I have no problem
      helping them with reagents, and I think it’s good to work with them (since they will probably have strong input into our NDA for a new anthrax vaccine), however, I don’t want to be just a “gofer” for someone who’s attempting to do what we are attempting to do. I like the idea of collaboration, but not parasitization. What are your thoughts? Could I write to and and ask them for a detailed plan with respect to what they want to do with the PA? (T6h)en we can decide if a meeting is needed. – Bruce
      >—–Original Message—– >From: >Sent: Thursday, July 26, 2001 11:42 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: MTA > >Bruce, > >I believe works for – yes? If so, they are interested in developing a potency assay in the mou(s6e), but we have not been in regular discussions with the group, and have only had a single meeting with them. I’d like to at least meet with them one more time before we commit to giving them any of our reagents. > > > > —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, July 20, 2001 11:19 AM > To: > Cc: > Subject: MTA > > > from us for some mouse immunization studies. I have written up an MTA request. Please look it over, and if you approve, please forward it to and > > > > > > >
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      of the FDA (CBER) has requested some rPA vaccine (Alhydrogel + rPA)
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      Thanks. – Bruce
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      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      RE: Covance Rabbits Monday, August 06, 2001 7:58:25 AM
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      Yes! They should go to
      >—–Original Message—– >From: >Sent: Monday, August 06, 2001 7:57 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: FW: Covance Rabbits > >Has anything been decided on these animals? > >—–Original Message—– >From: >Sent: Friday, August 03, 2001 3:36 PM >To: >Subject: Covance Rabbits > > I contacted Covance about the rabbits. I wanted to know if you were still interested in housing the rabbits at Covance. If you want to do this I can make the arrangements. I will need an APC to charge. If not they are scheduled to come in Thursday. Please let me know ASAP! Thank you! > >Scott
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      From: To: Cc: Subject: Date:
      I approve. – Bruce Ivins
      Ivins, Bruce E Dr USAMRIID
      RE: CRM Delivery Order 0004 Tuesday, August 07, 2001 9:15:57 AM
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      >—–Original Message—– >From: >Sent: Tuesday, August 07, 2001 8:55 AM >To: Ivins, Bruce E Dr USAMRIID >Cc: >Subject: CRM Delivery Order 0004 > >Dr. Ivins: > >I have received the invoice for for the month of July in the amount of $3,534.35. Please email reply to all your approval. Any questions, please call. >
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      From: To: Cc: Subject: Date:
      I approve. – Bruce Ivins
      Ivins, Bruce E Dr USAMRIID
      RE: CRM Delivery Order 0086 Tuesday, August 07, 2001 11:53:58 AM
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      >—–Original Message—– >From: >Sent: Tuesday, August 07, 2001 11:07 AM >To: Ivins, Bruce E Dr USAMRIID >Cc: >Subject: CRM Delivery Order 0086 > >Dr. Ivins: > >I have received the invoice for for the month of July in the amount of $4,969.09. Please email reply to all your approval. Any questions, please call. >
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      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      rPA meeting – 6 AUG 01 Tuesday, August 07, 2001 2:59:52 PM
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      PROPRIETARY – NOT FOR PUBLIC DISTRIBUTION
      (6) Here is what I have for the rPA meeting yesterday:
      1) Every other week there will be rPA product development team meetings.
      2) discussed the progress on his mouse potency assay studies. The rPA/Alhydrogel vaccines he has been formulating immediately prior to injection do not contain formaldehyde. The A/j mice in his first study were immunized intramuscularly with a single dose of vaccine containing 1, 3.162, 10, 31.62 or 100 micrograms of PA. Four weeks after immunization, the mice were challenged subcutaneously with 10 LD50 of Sterne spores. There was little difference among the doses with respect to antibody titers or protection, although the titers appeared to rise weekly. No information was available on the rabbit dose titrations. The next experiment will look at doses of 3.162, 1, .3, 0.1, and 0.03 micrograms of PA in the vaccines.
      3) Bruce talked about the status of stability studies (rPA vaccine with and without formaldehyde) in the rabbit. The rabbits will be held and immunized at Covance, then shipped to USAMRIID for challenge. The survival data should be in by the end of October. If there is enough available, some the the vaccines will be given to o physically characterize. He will try to present the data at the next meeting. will identify other items, such as sucrose, which would be acceptable as stabilizers in a new rPA vaccine. It was emphasized that we need to look at both physical stability and potency stability in a new rPA vaccine. The use of formaldehyde as a stabilizer in vaccines is discouraged by the FDA.
      4) is writing up an animal protocol to look at the potency of rPA vaccines containing different isof(o6r)ms. He will talk to Bruce about the protocol.
      5) discussed a study to be done at Battelle looking at the stability of rPA vaccine (no for(m6a)ldehyde,100microgramsPAper0.5-mldose).Battellewillwantatleast25mlofeachvaccine.
      6) said that wants to use a mixture of different monoclonal antibodies to PA as a sta(n6d)ard in assays. The FDA is working on a mouse potency test and has asked for a large amount of rPA. We will need to have a meeting concerning their plans and requests for reagents.
      7) talked about the progress of the Battelle contracted study. The first iteration of the subcutaneous challenge in rabbits has been done. With one dose of antiserum, all the animals died. With 2 doses of antiserum, all the animals survived. With respect to the non-human primates, an animal protocol should be submitted this week. Immunizations should start in a few weeks, with plasma phoresis every two weeks to obtain antiserum.
      8) talked about the problems with the TNA assay. The two sources of problems appear to be glu(t6a)mine breakdown in the tissue culture medium and the use of scrapers rather than glass beads to remove attached cells.
      9) said that the Phase I clinical study is planned for 18 months from now. It will be at the Un(iv6e)rsity of Maryland and run in cooperation with NIAID. Vaccine/Alhydrogel with PA doses of 5, 25, 50 and 100 micrograms of PA will be administered. In addition, doses of 25 and 100 micrograms of PA without Alhydrogel will be given. Acute toxicity studies will need to be done. Assays will be outsourced.
      10) At the next meeting Bruce will go over data on the adsorption of PA to Alhydrogel.
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      From: To: Subject: Date:
      Hi,
      Ivins, Bruce E Dr USAMRIID
      Isoform studies Tuesday, August 07, 2001 3:04:08 PM
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      I originally thought that our isoform studies would involve injections of vaccine at 0 and 4 weeks, then challenge at 10 weeks. However, if you think that we would be more likely to see differentiating results with one shot, we can do that. Please let me know. Otherwise we’ll continue with the 2 shots.
      – Bruce
      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      RE: Isoform studies Tuesday, August 07, 2001 7:32:43 PM
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      50 micrograms is what we are planning. – Bruce
      >—–Original Message—– >From: >Sent: Tuesday, August 07, 2001 4:36 PM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: Isoform studies > >What amount of rPA/dose? > > —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Tuesday, August 07, 2001 3:04 PM > To: > Subject: Isoform studies > > Hi, > weeks, then challenge at 10 weeks. However, if you think that we would be more likely to see differentiating results with one shot, we can do that. Please let me know. Otherwise we’ll continue with the 2 shots. > > – Bruce
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      I originally thought that our isoform studies would involve injections of vaccine at 0 and 4
      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      RE: Isoform studies Wednesday, August 08, 2001 9:56:44 AM
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      We haven’t set the challenge yet. For screening different vaccine candidates, we frequently use parenteral challenge, since we can tightly control the dose, do more animals at a time, and require fewer people and less labor to accomplish the task. In one day, we can do an aerosol challenge of 30 rabbits, employing about 8-10 people and 4 rooms (1 for animals, 2 hood lines, 1 spore and bacterial plating). In 1/2 day we can do a parenteral challenge of 60 rabbits, employing 2 people and 1 room. We use aerosol challenge when we have advanced our candidates along, as we are currently doing in
      protocol with the rPA/Alhydrogel/PBS. My preference at this stage would be a parenteral challenge, since it would give us the information we are looking for with respect to titers and protection differences. – Bruce
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      >—–Original Message—– >From: >Sent: Wednesday, August 08, 2001 8:34 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: Isoform studies > >Bruce, > >Will it be an aerosol challenge? > > —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Tuesday, August 07, 2001 7:33 PM > To: > Subject: RE: Isoform studies > > 50 micrograms is what we are planning. > – Bruce > > —–Original Message—– > From: > Sent: Tuesday, August 07, 2001 4:36 PM > To: Ivins, Bruce E Dr USAMRIID > Subject: RE: Isoform studies > > What amount of rPA/dose? > > —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Tuesday, August 07, 2001 3:04 PM > To: > Subject: Isoform studies > > Hi, > 0 and 4 weeks, then challenge at 10 weeks. However, if you think that we would be more likely to see differentiating results with one shot, we can do that. Please let me know. Otherwise we’ll continue with the 2 shots. > > – Bruce
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      I originally thought that our isoform studies would involve injections of vaccine at
      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      FW: Money into Battelle contract Wednesday, August 08, 2001 3:16:02 PM
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      >—–Original Message—– >From: >Sent: Wednesday, August 08, 2001 1:26 PM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: Money into Battelle contract > >Bruce: the way the contract works is th e following. You first need to write up a detailed SOW (statement of work). This is sent to Battelle through me. Battelle may call you to get further information or clarification on the SOW. They will send back a fixed cost for the work. If you approve their proposal which will include the cost, will do a form 9 and both will go froward (RAD IV and USAMRAA) for awarding. For every task order, a separate form 9 using your fund site(s) below will be used. It may sound complicated but it is not. If you want further explanation, come up and see me. > (6) >———- >From: Ivins, Bruce E Dr USAMRIID >Sent: Monday, August 6, 2001 12:30 PM >To: >Subject: Money into Battelle contract > >Hi, > (6C)ould we move some money from the following APCs into the Battelle contract for some anthrax protective antigen work?
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      > > > >Whatever I (we) need to do to accomplish this, please let me know. We would like to transfer the money as soon as possible.
      > >Thanks! > >- Bruce >
      a) From 6IDK – $17,000 b) From 6IDT – $13,000 c) From GDE – $75,000
      From: To: Subject: Date:
      Thanks, – Bruce (6)
      Ivins, Bruce E Dr USAMRIID
      RE: rPA meeting – 6 AUG 01 Wednesday, August 08, 2001 3:25:09 PM
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      >—–Original Message—– >From: >Sent: Wednesday, August 08, 2001 3:24 PM >To: Ivins, Bruce E Dr USAMRIID;
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      >Cc: >Subject: RE: rPA meeting – 6 AUG 01 > >Comments added in blue. > > > > > > >
      > DVC:
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      PROPRIETARY – NOT FOR PUBLIC DISTRIBUTION
      rPA RIID PDT Meeting August 6, 2001 0900 to 1030 Attendees: USAMRIID:
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      > > > > > rPA/Alhydrogel vaccines he has been formulating immediately prior to injection do not contain formaldehyde. The A/j mice in his first study were immunized intramuscularly with a single dose of vaccine containing 1, 3.162, 10, 31.62 or 100 micrograms of PA. Four weeks after immunization, the mice were challenged subcutaneously with 10 LD50 of Sterne spores. There was little difference among the doses with respect to antibody titers or protection, although the titers appeared to rise weekly. No information was available on the rabbit dose titrations. The next experiment (currently in progress) will look at doses of 3.162, 1, .3, 0.1, and 0.03 micrograms of PA in the vaccines. Control = PBS + Alhydrogel.
      JVAP: 1) Every other week there will be rPA product development team meetings. 2) discussed the progress on his mouse potency assay studies. The
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      > > 3) Bruce talked about the status of stability studies (rPA vaccine with and without formaldehyde) in the rabbit. The rabbits will be held and immunized at Covance, then shipped to USAMRIID for challenge. Bruce – did I understand correctly that these animals were being immunized w/ vaccine prepared ~ 5 years ago(evaluating the potency stability of these old preps)? The survival data should be in by the end of October. If there is enough available, some the the vaccines will be given to to evalutate protein stability over time by physical characterization . He will try to present the data at the next meeting with data obtained previously evaluating protein stability by SDS w/ and w/o formaldehyde. will identify other items, such as sucrose, which would be acceptable as stabilizers in a new rPA vaccine. It was emphasized that we need to look at both physical stability and potency stability in a new rPA vaccine. The use of formaldehyde as a stabilizer in vaccines is discouraged by the FDA. > > 4) s writing up an animal protocol to look at the potency of rPA vaccines containing the two differen(t6)isoforms. He will talk to Bruce about the protocol. > > 5) discussed a study to be done at Battelle looking at stability evaluated by both the biophysical inte(g6r)ity and the biological activity of the formulated rPA vaccine (no formaldehyde, 100
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      micrograms PA per 0.5-ml dose in isotonic buffer at pH 7.4 adsorbed to alhydrogel) stored at various temperatures over a period of time. Product to be used will be material prepared by BDP in 1998. Battelle will want at least 25 ml of each vaccine(this is 25 mL per experimental group of the vaccine prep at the 100 ug/.5mL dose I think.
      > > 6) said that wants to use a mixture of different monoclonal antibodies to PA as a standard in assays. The FDA is working on a mouse potency test and has asked for a large amount of rPA. USAMRIID will need to have a meeting concerning their plans and requests for reagents. > > 7) talked about the progress of the Battelle contracted study. The first iteration of the subcutaneous challenge in rabbits has been done. With one dose of antiserum, all the animals died. With 2 doses of antiserum, all the animals survived. Additional studies delayed pending resolution of the problems w/ the TNA assay. With respect to the non-human primates, an animal protocol should be submitted this week. Immunizations should start in a few weeks, with plasma phoresis every two weeks to obtain antiserum. > > 8) talked about the problems with the TNA assay. The two sources of problems appear to be g(lu6t)amine breakdown in the tissue culture medium and the use of scrapers rather than glass beads(reused and prepped by central glassware) to remove attached cells. > > 9) said that the Phase I clinical study is planned for 18 months from now. It will be at the University o(6f)Maryland and run in cooperation with NIAID. Vaccine/Alhydrogel with PA doses of 5, 25, 50 and 100 micrograms of PA will be administered. In addition, doses of 25 and 100 micrograms of PA without Alhydrogel will be given. Acute toxicity studies will need to be done. Assays will be outsourced. > > 10) At the next meeting Bruce will go over data on the adsorption of PA to Alhydrogel.
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      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID Ivins, Bruce E Dr USAMRIID FW: Notification of Protocol Completion/Termination Form due for protocol:B99-07 Friday, August 10, 2001 8:53:47 AM
      —–Original Message—– From: Sent: Wednesday, August 08, 2001 3:25 PM To: Ivins, Bruce E Dr USAMRIID Subject: Notification of Protocol Completion/Termination Form due for protocol:B99-07
      REPORT OF PROTOCOL COMPLETION / TERMINATION
      Date : 08/08/2001 Division : BACTERIOLOGY Investigator : IVINS, BRUCE Protocol : B99-07
      TITLE : Protective Effect of Immunostimulatory Oligonucleotides Against Bacillus anthracis Intramuscular Challenge in Guinea
      1. Was the protocol been completed as orginally designed and/or ammemded? Explain: Yes
      2. Date completed: _25 JUN 01 If terminated prior to completion, explain:
      3. Report any change to the number of animals in the painful procedure categories if different than the number reported in the approved protocol.
      No Pain = 48 (30%) Pain = 112 (70%) Originally we believed that 50% of the animals would be in the NO PAIN category
      4. What were the results and/or conclusions? (Report any negative findings if applicable.)
      The results indicated that specific immunity to anthrax spore challenge in guinea pig is enhanced by coadministration of CpG oligonucleotides.
      5. Will the research result in a publication or presentation? (Provide specifics.) We intend to present the data at the Annual Meeting of the
      American Society for Microbiology, Salt Lake City, Utah, in 2002.
      Date : 10 AUG 01 Signature/Division : __Bruce E. Ivins, USAMRIID Bacteriology Division_
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      RETURN THIS FORM TO VETERINARY MEDICINE DIVISION
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      From: To: Subject: Date: Attachments:
      Ivins, Bruce E Dr USAMRIID
      RE: Isoform Rabbit protocol Friday, August 10, 2001 10:01:44 AM
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      – I have suggested some corrections. Has seen this and added his input? Have you done (a6)search at the Library? Has this been taken to the appropriate vet for comments yet? I can help you
      with each when you have time. – Bruce – Below is my corrected version
      >—–Original Message—– >From: >Sent: Thursday, August 09, 2001 3:39 PM >To: Ivins, Bruce E Dr USAMRIID >Cc:
      >Subject: Isoform Rabbit protocol > >Hi Bruce, >Attached are my mods to your rabbit protocol to address protection by the pure isoforms. I need your input for statistical numbers expected when using 16 rabbits per group. Thanks in advance.
      > <>
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      From: To: Subject: Date: Attachments:
      Ivins, Bruce E Dr USAMRIID
      FW: New Sporulation Medium Monday, August 13, 2001 12:00:47 PM
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      >—–Original Message—– >From: >Sent: Monday, August 13, 2001 11:51 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: New Sporulation Medium > >Please find attached the recipes for preparation of both media. I suggest cutting the NSM recipe in half for I only have ten plates for you to use. Let me know when you want to pick up the Lab Lemco from me. > >—–Original Message—– >From: Ivins, Bruce E Dr USAMRIID >Sent: Monday, August 13, 2001 8:27 AM >To: >Subject: New Sporulation Medium >
      > > We have a strain of B. anthracis that we are having trouble with making spores. Can you give us the formula for NSM (new sporulation medium)? Also, if you have the formula for any other good spore formation media, we’d appreciate knowing about it. > >Thanks! > >- Bruce
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      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      rPA vaccine preparation Wednesday, August 15, 2001 8:59:45 AM
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      rPA vaccine per 0.5-ml dose:
      50 micrograms rPA 0.047 ml of Alhydrogel (2% aluminum oxide) containing 0.5 mg of metallic aluminum Dulbeccos PBS (pH 7.4) without calcium or magnesium, qs to 0.5 ml
      First add PBS then Alhydrogel. Mix. Then add rPA and mix. Allow to adsorb for 1 hour (or longer). – Bruce
      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      rPA vaccine preparation Wednesday, August 15, 2001 8:59:45 AM
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      rPA vaccine per 0.5-ml dose:
      50 micrograms rPA 0.047 ml of Alhydrogel (2% aluminum oxide) containing 0.5 mg of metallic aluminum Dulbeccos PBS (pH 7.4) without calcium or magnesium, qs to 0.5 ml
      First add PBS then Alhydrogel. Mix. Then add rPA and mix. Allow to adsorb for 1 hour (or longer). – Bruce
      From: To: Cc: Subject: Date:
      Ivins, Bruce E Dr USAMRIID Ivins, Bruce E Dr USAMRIID;
      RE: Dugway Wednesday, August 15, 2001 1:23:52 PM
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      Hi, we’ll be ready to go here. Hope you had a great vacation! – Bruce
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Wednesday, July 25, 2001 9:28 AM
      (6)Any word on the Spore contract? Just as soon as everything is approved and signed on all sides,
      To: ‘ Cc: Subject: RE: Dugway
      Thanks, I’m on vacation for a week, until August 1. If you have information, please forward it to
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      – Bruce
      and as well. Talk to you later!
      —–Original Message—– From: Sent: Wednesday, July 25, 2001 9:35 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Dugway
      Hi Bruce, I’m just getting back today from being gone for three weeks. I’ll
      track it down as I catch up today and tomorrow and let you know where we stand. Good to hear from you.
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      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Tuesday, July 17, 2001 6:01 AM To: Subject: FW: Dugway
      ,
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      (b)
      (b) (6)
      (b)
      How are things going on your end with respect to the contract? According to our contract person, (below) it was sent and should be there. I’m looking forward to working with you again on these. – Bruce
      (6)
      > —–Original Message—– > From: > Sent: Tuesday, July 10, 2001 10:16 AM > To: Ivins, Bruce E Dr USAMRIID
      (b) (6)
      (b) (6)
      > Subject: Dugway > > Bruce, >
      > Money and statement of work went to Dugway on Monday. They confirmed receipt. > >
      > > (6) >
      (b)
      From: To: Cc: Subject: Date:
      Ivins, Bruce E Dr USAMRIID Ivins, Bruce E Dr USAMRIID;
      RE: Dugway Wednesday, August 15, 2001 1:23:52 PM
      (b) (6)
      (b) (6)
      (b)
      Hi, we’ll be ready to go here. Hope you had a great vacation! – Bruce
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Wednesday, July 25, 2001 9:28 AM To: Cc: Subject: RE: Dugway
      Thanks, I’m on vacation for a week, until August 1. If you have information, please forward it to
      (6)Any word on the Spore contract? Just as soon as everything is approved and signed on all sides,
      (b) (6)
      (b) (6)
      (b)
      (b) (6) (6)
      (b) (6)
      – Bruce
      and as well. Talk to you later!
      —–Original Message—– From: Sent: Wednesday, July 25, 2001 9:35 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Dugway
      Hi Bruce, I’m just getting back today from being gone for three weeks. I’ll
      track it down as I catch up today and tomorrow and let you know where we stand. Good to hear from you.
      (6)
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Tuesday, July 17, 2001 6:01 AM To: Subject: FW: Dugway
      (6) How are things going on your end with respect to the contract? According to our contract person, (below) it was sent and should be there. I’m looking forward to working with you again on these. – Bruce
      > —–Original Message—– > From: > Sent: Tuesday, July 10, 2001 10:16 AM > To: Ivins, Bruce E Dr USAMRIID
      (b) (6)
      (b)
      (b) (6)
      (b)
      (b) (6)
      (b) (6)
      > Subject: Dugway > > Bruce, >
      > Money and statement of work went to Dugway on Monday. They confirmed receipt. > >
      > > (6) >
      (b)
      From: To: Subject: Date:
      Ivins, Bruce E Dr USAMRIID
      FW: Dugway Thursday, August 16, 2001 10:13:54 AM
      (b) (6)
      (b)
      (b)
      and – As soon as I hear I’ll get the info to you. Hopefully we’ll get started soon to get the (n6e)w spor(e6s) made and purified.
      – Bruce
      —–Original Message—– From: Sent: Wednesday, August 15, 2001 7:19 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Dugway
      not word to me yet. I’ll be back next Wed and will hound dog it and get an answer.
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Wednesday, August 15, 2001 11:24 AM To: Ivins, Bruce E Dr USAMRIID; Cc: Subject: RE: Dugway
      Hi,
      (6)Any word on the Spore contract? Just as soon as everything is approved and signed on all sides, we’ll be ready to go here. Hope you had a great vacation!
      – Bruce
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Wednesday, July 25, 2001 9:28 AM
      (b) (6)
      (b) (6)
      (b) (6)
      (b)
      To: ‘ Cc: Subject: RE: Dugway
      Thanks, I’m on vacation for a week, until August 1. If you have informat(io6n), please forward it to and as well. Talk to you later! – Bruce
      —–Original Message—– From: Sent: Wednesday, July 25, 2001 9:35 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Dugway
      Hi Bruce, I’m just getting back today from being gone for three weeks. I’ll
      track it down as I catch up today and tomorrow and let you know where we stand. Good to hear from you.
      (b) (6)
      (b) (6)
      (b)
      (b) (6)
      (b) (6)
      (b) (6)
      (b)
      (6)
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Tuesday, July 17, 2001 6:01 AM To: Subject: FW: Dugway
      (6) How are things going on your end with respect to the contract? According to our contract person, (below) it was sent and should be there. I’m looking forward to working with you again on these. – Bruce
      > —–Original Message—–
      > From: > Sent: Tuesday, July 10, 2001 10:16 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: Dugway > > Bruce, > > Money and statement of work went to Dugway on Monday. They confirmed receipt. > > > > (6) >
      (b) (6)
      (b)
      (b) (6)
      (b) (6)
      (b)
      From: To: Subject: Date:
      FYI, – Bruce
      (6)
      Ivins, Bruce E Dr USAMRIID
      FW: Dugway Thursday, August 16, 2001 10:35:02 AM
      (b) (6)
      (b)
      —–Original Message—– From: Sent: Thursday, August 16, 2001 10:34 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Dugway
      Bruce,
      MIPR was sent July 9 and a confirmation response received. Seems strange that an acknowledgement hasn’t made its way to the technical POC by now. Maybe he should call their contracting office.
      (6)
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Thursday, August 16, 2001 10:14 AM To: Subject: FW: Dugway
      and – As soon as I hear I’ll get the info to you. Hopefully we’ll get started soon to get the (n6e)w spor(e6s) made and purified.
      – Bruce
      —–Original Message—– From: Sent: Wednesday, August 15, 2001 7:19 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Dugway
      not word to me yet. I’ll be back next Wed and will hound dog it and get an answer.
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Wednesday, August 15, 2001 11:24 AM
      To: Ivins, Bruce E Dr USAMRIID; ‘ Cc: Subject: RE: Dugway
      (b) (6)
      (b) (6)
      (b)
      (b) (6)
      (b)
      (b)
      (b) (6)
      (b) (6)
      Hi, , (6)Any word on the Spore contract? Just as soon as everything is
      approved and signed on all sides, we’ll be ready to go here. Hope you had a great vacation!
      – Bruce
      (b) (6)
      (b)
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Wednesday, July 25, 2001 9:28 AM
      To: ‘ Cc: Subject: RE: Dugway
      Thanks, I’m on vacation for a week, until August 1. If you have informat(io6n), please forward it to and as well. Talk to you later! – Bruce
      —–Original Message—– From: Sent: Wednesday, July 25, 2001 9:35 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Dugway
      Hi Bruce, I’m just getting back today from being gone for three weeks. I’ll
      track it down as I catch up today and tomorrow and let you know where we stand. Good to hear from you.
      (6)
      —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Tuesday, July 17, 2001 6:01 AM To: Subject: FW: Dugway
      b(6) How are things going on your end with respect to the contract? )According to our contract person, (below) it was sent and should (be there. I’m looking forward to working with you again on these.
      (b) (6)
      (b) (6)
      (b)
      (b) (6)
      (b) (6)
      (b) (6)
      (b)
      (b) (6)
      (b)
      (b) (6)
      – Bruce
      6 )
      > —–Original Message—– > From: > Sent: Tuesday, July 10, 2001 10:16 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: Dugway > > Bruce, > > Money and statement of work went to Dugway on Monday. They confirmed receipt. > > > > (6) >
      (b) (6)
      (b)

      • DXer said

        From: To: Cc: Subject: Date: Attachments: Ivins, Bruce E Dr USAMRIID FY 2001 Progress info Tuesday, August 21, 2001 8:06:25 AM (b) (6) (b) (6) (b) (6) Here is my FY 2001 progress: – Bruce From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID FW: rPA meeting – 6 AUG 01 Wednesday, August 22, 2001 7:34:27 AM (b) (6) (b) (6) Chief…Sir… – Here are the corrected minutes. did such a good job of suggesting changes, do you want to ask her if she would like to take minutes tomorrow? If not, I’ll do them again. – your humble servant – Bruce >—–Original Message—– >From: >Sent: Wednesday, August 08, 2001 3:24 PM >To: Ivins, Bruce E Dr USAMRIID; b >Cc: ) >Subject: RE: rPA meeting – 6 AUG 01 ( (b) (6) (b) (6) (b) (6) (b)( (6) (b) (6) > >Thanks to for all of her helpful comments! > > > PROPRIETARY – NOT FOR PUBLIC DISTRIBUTION > > rPA RIID PDT Meeting > August 6, 2001 0900 to 1030 > Attendees: 6 ) (b) (6) (b) (6) (b) (6) > > 1) Every other week there will be rPA product development team meetings. > > 2) discussed the progress on his mouse potency assay studies. The rPA/Alhydrogel vaccines he has been formulating immediately prior to injection do not contain formaldehyde. The A/j mice in his first study were immunized intramuscularly with a single dose of vaccine containing 1, 3.162, 10, 31.62 or 100 micrograms of PA. Four weeks after immunization, the mice were challenged subcutaneously with 10 LD50 of Sterne spores. There was little difference among the doses with respect to antibody titers or protection, although the titers appeared to rise weekly. No information was available on the rabbit dose titrations. The next experiment, currently in progress, will look at doses of 3.162, 1, .3, 0.1, and 0.03 micrograms of PA in the vaccines. The control in the experiments is PBS + Alhydrogel. > > 3) Bruce talked about the status of stability studies (rPA vaccine with and without formaldehyde) in the rabbit. The rabbits will be held and immunized at Covance, then shipped to USAMRIID for challenge. These animals are being immunized with vaccine prepared 5 years ago. so we are thus evaluating the potency stability of older vaccine preparations. The survival data should be in by the end of October. If there is enough available, some the the vaccines will be given to to evalutate protein stability over time by physical characterization . He will try to present the data at the next meeting with data obtained previously evaluating protein stability by SDS with and without formaldehyde. will identify other items, such as sucrose, which would be acceptable as stabilizers in a new rPA vaccine. It was emphasized that we need to look at both physical stability and potency stability in a new rPA vaccine. The use of formaldehyde as a stabilizer in vaccines is discouraged by the FDA. > > 4) is writing up an animal protocol to look at the potency of rPA vaccines containing the two different is(6of)orms. He will talk to Bruce about the protocol. > (b) (6) (b) (6) (b) (6) (b) > 5) discussed a study to be done at Battelle looking at stability evaluated by both the biophysica(6l)integrity and the biological activity of the formulated rPA vaccine (no formaldehyde, 100 micrograms PA per 0.5-ml dose in isotonic buffer at pH 7.4 adsorbed to alhydrogel) stored at various temperatures over a period of time. Product to be used will be material prepared by BDP in 1998. Battelle will want at least 25 ml of each vaccine, 25 mL per experimental group of the vaccine prep at the 100 ug/0.5mL dose. > > 6) said that wants to use a mixture of different monoclonal antibodies to PA as a stand(6a)rd in assays. The FDA is working on a mouse potency test and has asked for a large amount of rPA. USAMRIID will need to have a meeting concerning their plans and requests for reagents. > > 7) talked about the progress of the Battelle contracted study. The first iteration of the subcutaneous challenge in rabbits has been done. With one dose of antiserum, all the animals died. With 2 doses of antiserum, all the animals survived. Additional studies delayed pending resolution of the problems with the TNA assay. With respect to the non-human primates, an animal protocol should be submitted this week. Immunizations should start in a few weeks, with plasma phoresis every two weeks to obtain antiserum. > > 8) talked about the problems with the TNA assay. The two sources of problems appear to be glutam(6in)e breakdown in the tissue culture medium and the use of scrapers rather than glass beads (reused and prepped by central glassware) to remove attached cells. > > 9) said that the Phase I clinical study is planned for 18 months from now. It will be at the University(6o)f Maryland and run in cooperation with NIAID. Vaccine/Alhydrogel with PA doses of 5, 25, 50 and 100 micrograms of PA will be administered. In addition, doses of 25 and 100 micrograms of PA without Alhydrogel will be given. Acute toxicity studies will need to be done. Assays will be outsourced. > > 10) At the next meeting will go over data on the adsorption of PA to Alhydrogel. (6) (b) (b) (6) (b) (6) (b) (b) (b) (b) From: Ivins, Bruce E Dr USAMRIID To: “(b)(6) A Cc: Ivins, Bruce E Dr USAMRIID ; Subject: RE: MTA for rPA Date: Thursday, August 23, 2001 11:55:09 AM I am also going to talk to about this, since he is the head of the rPA vaccine program here. – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 11:33 AM (b) (6) (b) (6) (b) (6) To: ‘ Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA After a period of procrastination, for which I apologize, I finally managed to make my comments to your MTA for rPA. Could all of you please look into them and give us your feedback? Thanks for your help. —–Original Message—– From: Sent: Thursday, July 26, 2001 8:43 AM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) To: Cc: ‘ Ivins, Bruce E Dr USAMRIID Subject: MTA for rPA Good morning, I received your e-mail and the MTA attachments. I’ve completed the MTA template that USAMRIID uses. You’ll notice that some language is different. I would appreciate if you would review the document. If CBER/FDA accepts this MTA, I would appreciate if the responsible party would sign three of the agreements and send them to the below address. Unless I hear differently from you I will assume that this meets with your approval. Respectfully, Please send signed copies to this address: (b) (6) (b) (6) (b) (6) <> From: To: Cc: Subject: Date: Ivins, Bruce E Dr USAMRIID Ivins, Bruce E Dr USAMRIID; RE: MTA for rPA Thursday, August 23, 2001 11:55:09 AM A (b) (6) (b) (6) (b) (6) I am also going to talk to about this, since he is the head of the rPA vaccine program here. – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 11:33 AM (b) (6) To: ‘ Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA After a period of procrastination, for which I apologize, I finally managed to make my comments to your MTA for rPA. Could all of you please look into them and give us your feedback? Thanks for your help. —–Original Message—– From: Sent: Thursday, July 26, 2001 8:43 AM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) To: Cc: ‘ Ivins, Bruce E Dr USAMRIID Subject: MTA for rPA Good morning, I received your e-mail and the MTA attachments. I’ve completed the MTA template that USAMRIID uses. You’ll notice that some language is different. I would appreciate if you would review the document. If CBER/FDA accepts this MTA, I would appreciate if the responsible party would sign three of the agreements and send them to the below address. Unless I hear differently from you I will assume that this meets with your approval. Respectfully, Please send signed copies to this address: (b) (6) (b) (6) (b) (6) <> From: To: Subject: Date: Attachments: Ivins, Bruce E Dr USAMRIID FW: MTA for rPA Thursday, August 23, 2001 11:58:08 AM (b) (6) (b) (6) Hi, generate mouse antiserum. – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 11:33 AM (b) (6) (b) (6) I am forwarding this to you.This deals with the request by and at FDA for rPA to (b) (6) To: ‘ Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA After a period of procrastination, for which I apologize, I finally managed to make my comments to your MTA for rPA. Could all of you please look into them and give us your feedback? Thanks for your help. —–Original Message—– From: Sent: Thursday, July 26, 2001 8:43 AM (b) (6) (b) (6) (b) (6) (b) (6) To: Cc: ‘ Ivins, Bruce E Dr USAMRIID Subject: MTA for rPA Good morning, I received your e-mail and the MTA attachments. I’ve completed the MTA template that USAMRIID uses. You’ll notice that some language is different. I would appreciate if you would review the document. If CBER/FDA accepts this MTA, I would appreciate if the responsible party would sign three of the agreements and send them to the below address. Unless I hear differently from you I will assume that this meets with your approval. Respectfully, Please send signed copies to this address: (b) (6) (b) (6) (b) (6) (b) (6) <> (b) (6) From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID RE: MTA for rPA Thursday, August 23, 2001 11:59:51 AM (b) (6) (b) (6) has some problems with this whole thing. I think we’ll have to hold off for a bit as far as everyone signing until he is satisfied that we are simply not rPA suppliers to our competitors. – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 11:33 AM (b) (6) To: ‘ Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA After a period of procrastination, for which I apologize, I finally managed to make my comments to your MTA for rPA. Could all of you please look into them and give us your feedback? Thanks for your help. —–Original Message—– From: Sent: Thursday, July 26, 2001 8:43 AM (b) (6) (b) (6) (b) (6) (b) (6) To: Cc: ‘ Ivins, Bruce E Dr USAMRIID Subject: MTA for rPA Good morning, I received your e-mail and the MTA attachments. I’ve completed the MTA template that USAMRIID uses. You’ll notice that some language is different. I would appreciate if you would review the document. If CBER/FDA accepts this MTA, I would appreciate if the responsible party would sign three of the agreements and send them to the below address. Unless I hear differently from you I will assume that this meets with your approval. Respectfully, (b) (6) (b) (6) <> From: To: Cc: Subject: Date: Ivins, Bruce E Dr USAMRIID RE: MTA for rPA Thursday, August 23, 2001 12:18:17 PM (b) (6) (b) (6) (b) (6) Can we have a meeting about this? With so much rPA involved, perhaps a Cooperative Research and Development Agreement (CRDA) would be more appropriate than a Material Transfer Agreement (MTA). Please let us know what days and times are good for you, and if you would be able to make it up here again. Thanks. – Bruce —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Thursday, August 23, 2001 11:55 AM To: Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA (b) (6) (b) (6) I am also going to talk to about this, since he is the head of the rPA vaccine program here. – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 11:33 AM To: Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA After a period of procrastination, for which I apologize, I finally managed to make my comments to your MTA for rPA. Could all of you please look into them and give us your feedback? Thanks for your help. —–Original Message—– From: Sent: Thursday, July 26, 2001 8:43 AM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) To: Cc: ‘ Ivins, Bruce E Dr USAMRIID Subject: MTA for rPA Good morning, I received your e-mail and the MTA attachments. I’ve completed the MTA template that USAMRIID uses. You’ll notice that some language is different. I would appreciate if you would review the document. If CBER/FDA accepts this MTA, I would appreciate if the responsible party would sign three of the agreements and send them to the below address. Unless I hear differently from you I will assume that this meets with your (b) (6) approval. Respectfully, <> (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: FW: MTA for rPA Date: Thursday, August 23, 2001 12:31:24 PM (6) Just got this from The sooner we can meet, the better, I think. Do you want to respond to him, or shall I? – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 12:28 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: MTA for rPA Bruce, Thanks for writing. I’m sure that we can find a time to meet with you at Detrick. Unfortunately s on leave. He’ll be back at work on the 7th of September. I’ll talk to him about this. In the mean term, do you think we can get going? We have finalized our dosing experiment. I owe you the results, but I was waiting for the last set, to send you a comprehensive table. Most likely, next week. Our expectation was to start the “commercial” manufacturing of serum in September. Our contractor (Biocon) is ready to go. Anyway, I agree that probably the most efficient way to address all issues would be a meeting. I’ll make arrangements for it to happen. Regards. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, August 23, 2001 12:18 PM To: Cc: Subject: RE: MTA for rPA Can we have a meeting about this? With so much rPA involved, perhaps a Cooperative Research and Development Agreement (CRDA) would be more appropriate than a Material Transfer Agreement (MTA). Please let us know what days and times are good for you, and if you would be able to make it up here again. Thanks. – Bruce —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Thursday, August 23, 2001 11:55 AM To: Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA (b) (6) (b) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) I am also going to talk to about this, since he is the head of the rPA vaccine program here. – Bruce —–Original Message—– From: ] Sent: Thursday, August 23, 2001 11:33 AM To: Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: MTA for rPA After a period of procrastination, for which I apologize, I finally managed to make my comments to your MTA for rPA. Could all of you please look into them and give us your feedback? Thanks for your help. —–Original Message—– From: Sent: Thursday, July 26, 2001 8:43 AM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) To: Cc: ‘ Ivins, Bruce E Dr USAMRIID Subject: MTA for rPA Good morning, I received your e-mail and the MTA attachments. I’ve completed the MTA template that USAMRIID uses. You’ll notice that some language is different. I would appreciate if you would review the document. If CBER/FDA accepts this MTA, I would appreciate if the responsible party would sign three of the agreements and send them to the below address. Unless I hear differently from you I will assume that this meets with your approval. Respectfully, <> (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: FW: rPA vaccine Date: Saturday, August 25, 2001 11:38:34 AM Can you give me some guidance on this? I personally don’t think that we will be able to supply him the large amount of PA that he has requested before the middle of September. – Bruce —–Original Message—– From: Sent: Friday, August 24, 2001 5:03 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: rPA vaccine Bruce, We set a contract with Biocon on the basis of the e-mail exchange below. Unfortunately, as I explained in a prior communication, won’t be back till September 7. I can probably ask the people of Biocon to delay the starting date of the project; however, I’m a little apprehensive about funding, due to end of the fiscal year issues. Do you think we can count on having the material within the first two weeks of September? Otherwise, I may be forced to cancel the contract, which I would prefer not to do. One more thing I can attempt to do is to arrange a conference call connecting Bruce from somewhere in the US. Do you want me to do that? Please explore with such a possibility. Thanks. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Monday, July 09, 2001 7:47 AM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) To: ‘ Ivins, Bruce E Dr USAMRIID Cc: Subject: RE: rPA vaccine Hi, (6)We will be happy to prepare more vaccine for you and have it ready for you on Jul 19. In addition, we can supply you with the vaccine for the 500 mice. The PA we have been using is the GLP/GMP PA produced at the MARP here at Fort Detrick. It is very high quality, but the supplies are becoming somewhat limited. Thus, if more PA vaccine is needed later, we may need to talk about your specific needs. (We have a great deal of PA which is not GLP/GMP, which is active in toxin neutralization tests, and which is protectively immunogenic. It’s just not the GLP/GMP material that we have used in the past.) Also, if more material is desired past that which is needed for the 500 mice, We should probably establish an MTA or a CRDA. Please let me know a little in advance exactly when you or your wife will be coming, so we can be sure to be here at that time. Thank you. – Regards, – Bruce —–Original Message—– From: ] Sent: Friday, July 06, 2001 3:23 PM To: ‘Bruce.Ivins@DET.AMEDD.ARMY.MIL’ (b) (6) (b) (6) (b) (b) (6) Cc: Subject: rPA vaccine Dear Bruce, Thanks again for providing us with vaccine for our pilot study of rPA immunogenicity. We have gotten already some results that I’ll share with you as soon as they are in presentable shape. The next (third) immunization of the mice with rPA is scheduled for July 20. I’ll be out of town Monday through Wednesday of that week. Therefore, I want to ask you if either my wife or I can pick up the vaccine on Thursday, July 19. I remind you that in this instance groups of 10 mice will be immunized with 1, 3 and 9 micrograms per 0.5 ml dose, and that we will inject ten controls with 0.5 ml of adjuvant in saline. In addition, I want to explore the possibility of enrolling your help in preparing the big batch of mouse anti-PA serum that is the final goal of this preliminary study. We are in the process of establishing a contract to get 500 mice immunized up to three times with a dose chosen on the basis of the results of the pilot study. I don’t anticipate the need of using the 18, 18, 9 micrograms schedule. Also we are not certain yet if a third dose will be necessary. Summarizing, do you think that USAMRIID (you) could provide us with up to 2 X 500 dose of vaccine (6 micrograms in 0.5 ml) plus 500 doses (3 micrograms in 0.5 ml). According to my calculations a total of 7.5 mg of rPA would be needed. As usual, a little excess would be necessary to ensure appropriate dosing. In addition of the acknowledgement of your contribution to the preparation of this material and access to our results, we are willing to share a fraction of the serum produced. I would like to hear your opinion about this project. We do not expect that the immunization of the 500 animals will begin before the conclusion of our pilot study, approximately on August 17. However, we should finish contract writing for the project as soon as possible, due to end of the fiscal year deadlines. Please let me know what do you think. Regards. (b) (6) From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID “(b) (6) RE: rPA vaccine Saturday, August 25, 2001 11:55:33 AM (b) (6) I doubt that a CRDA can be signed by 15 SEP. The PA cost us approximately $1,000 per vial, and several vials will be needed for the mouse immunizations. believes that this should be a cooperative agreement, rather than an MTA, and that we need a meeting first. If you can meet with us at 10 am on September 3, I believe that we can straighten things out pretty much. I’m not sure how long a contract can be delayed there. Here at USAMRIID, we can extend payment past the FY until 31 DEcember. Can you meet here at 10 am on 3 September? – Bruce —–Original Message—– From: Sent: Friday, August 24, 2001 5:03 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: rPA vaccine Bruce, We set a contract with Biocon on the basis of the e-mail exchange below. Unfortunately, as I explained in a prior communication, won’t be back till September 7. I can probably ask the people of Biocon to delay the starting date of the project; however, I’m a little apprehensive about funding, due to end of the fiscal year issues. Do you think we can count on having the material within the first two weeks of September? Otherwise, I may be forced to cancel the contract, which I would prefer not to do. One more thing I can attempt to do is to arrange a conference call connecting Bruce from somewhere in the US. Do you want me to do that? Please explore with such a possibility. Thanks. (6) —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Monday, July 09, 2001 7:47 AM (b) (6) (b) (6) (b) (6) (b) To: ‘ Ivins, Bruce E Dr USAMRIID Cc: Subject: RE: rPA vaccine Hi, (6)We will be happy to prepare more vaccine for you and have it ready for you on Jul 19. In addition, we can supply you with the vaccine for the 500 mice. The PA we have been using is the GLP/GMP PA produced at the MARP here at Fort Detrick. It is very high quality, but the supplies are becoming somewhat limited. Thus, if more PA vaccine is needed later, we may need to talk about your specific needs. (We have a great deal of PA which is not GLP/GMP, which is active in toxin neutralization tests, and which is protectively immunogenic. It’s just not the GLP/GMP material that we have used in the past.) Also, if more material is desired past that which is needed for the 500 mice, We should probably establish an MTA or a CRDA. Please let me know a little in advance exactly when you or your wife will be coming, so we can be sure to be here at that time. Thank you. – Regards, (b) (6) (b) (6) (b) – Bruce —–Original Message—– From: Sent: Friday, July 06, 2001 3:23 PM To: ‘Bruce.Ivins@DET.AMEDD.ARMY.MIL’ Cc: Subject: rPA vaccine Dear Bruce, Thanks again for providing us with vaccine for our pilot study of rPA immunogenicity. We have gotten already some results that I’ll share with you as soon as they are in presentable shape. The next (third) immunization of the mice with rPA is scheduled for July 20. I’ll be out of town Monday through Wednesday of that week. Therefore, I want to ask you if either my wife or I can pick up the vaccine on Thursday, July 19. I remind you that in this instance groups of 10 mice will be immunized with 1, 3 and 9 micrograms per 0.5 ml dose, and that we will inject ten controls with 0.5 ml of adjuvant in saline. In addition, I want to explore the possibility of enrolling your help in preparing the big batch of mouse anti-PA serum that is the final goal of this preliminary study. We are in the process of establishing a contract to get 500 mice immunized up to three times with a dose chosen on the basis of the results of the pilot study. I don’t anticipate the need of using the 18, 18, 9 micrograms schedule. Also we are not certain yet if a third dose will be necessary. Summarizing, do you think that USAMRIID (you) could provide us with up to 2 X 500 dose of vaccine (6 micrograms in 0.5 ml) plus 500 doses (3 micrograms in 0.5 ml). According to my calculations a total of 7.5 mg of rPA would be needed. As usual, a little excess would be necessary to ensure appropriate dosing. In addition of the acknowledgement of your contribution to the preparation of this material and access to our results, we are willing to share a fraction of the serum produced. I would like to hear your opinion about this project. We do not expect that the immunization of the 500 animals will begin before the conclusion of our pilot study, approximately on August 17. However, we should finish contract writing for the project as soon as possible, due to end of the fiscal year deadlines. Please let me know what do you think. Regards. (b) (6) (b) (6) From: To: Subject: Date: Hi, Ivins, Bruce E Dr USAMRIID RE: spore work Saturday, August 25, 2001 7:01:03 PM (b) (6) (b) (6)That’s great news. If you want to initiate an EA101, then we’ll get the spores ready on this end. Let’s plan on sending them on Tuesday, September 4. That should give time to everyone on both ends. We’ll send you spores from the previous spores you sent us. They will be on gel ice. We’ll send you 1 ml, 3 X 10^10 per ml, in a Nunc polypropylene vial. If you need more, let me know. We’ll wait for the EA101. My FAX is who is in charge of safety here, has a FAX number of Our facility registration number is – Thanks again! – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 7:00 PM To: Ivins (E-mail) Cc: Subject: spore work Bruce, I just received the test execution directive from the Program Analysis Office for the spore preparation. We’re good to go. Did you want to send a stock source of AMES for us to use? If you do, then I’ll initiate and EA 101 on Monday or whenever you want to do it. Regards and have a good weekend, (b) (6) (b) (6) (b) (6) (b) (2) (b) (6) (b) (6) (b) (6) From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID “(b) (6) RE: Anthrax, mice, and CpG Saturday, August 25, 2001 7:06:43 PM (b) (6) I can get it to you this week, Please tell me the day you are planning to immunize. Also, how many doses of each (AVA and rPA vaccine) will you need. (Include any extra that you wish to have.) Thanks! – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 4:43 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Hope you are well. The monkeys were immunized and bled. We need the second dose of both types of vaccine for the booster immunization ASAP. We’ll re-bleed a few times, then send you the serum for testing. All the best, —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, July 19, 2001 6:27 PM (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG Hi, If you can get us 1 ml of serum, then we can probably go with that. – Bruce —–Original Message—– From: Sent: Thursday, July 19, 2001 5:41 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, I spoke too soon. Given that we plan to bleed the animals every 2 weeks, the vets won’t allow us to draw 5 ml each time (to get 2 ml of serum). Especially since we also want to do some serum chemistry and hematology on these animals. Can you make do with less serum? I’m only guessing, but when we studied mouse serum for anti-PA Abs by ELISA, the assay only took a few microliters (b) (6) (b) (6) (b) (6) (b) (6) of serum. More serum was needed for the neutralization assays, but if memory serves, the total volume required was approximately 300 ul. Thus, if you could get by with 0.5 – 1 ml of serum, everything will be OK. Otherwise, let me know, and we’ll figure something out. —–Original Message—– From: Sent: Thursday, July 19, 2001 4:08 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Will do. At least two 1 ml tubes of serum will be frozen down from each animal at each time point. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, July 19, 2001 3:11 PM (b) (6) (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG , We draw blood into serum separator tubes, draw off the serum after clotting, aliquot the serum as we wish, then freeze it. For small (about 1 ml or less) volumes, we use cryotubes. For larger volumes, we use 15-ml, screw-capped, conical polypropylene tubes. For bleed-outs (which I believe won’t be done in this experiment), we use 50-ml, screw-capped, conical polypropylene tubes. For each time-point in each monkey, we would like 2 aliquots of 1 ml of serum (thus, 2 ml total). That is enough to cover both ELISA and TNA tests. After all the samples are taken, I can drive down and pick up the frozen serum, bring it back, and the ELISA and TNA assays will be run. Thanks. I’m eagerly looking forward to this experiment. – Bruce —–Original Message—– From: Sent: Thursday, July 19, 2001 1:57 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, We’re all set for the monkey study. The animals are scheduled to be immunized on Tuesday, the day after the vaccine arrives. If memory serves, we’re injecting 0.5 ml intramuscularly. They’ll be re-immunized one month later. Bloods will be drawn every two weeks, and serum stored for you to test. In terms of collecting and storing the serum, I could use your advice. We do many of our ELISA assays in microtiter plates. If the same is true at your end, I could store the serum samples from each bleed in microtiter (b) (6) (b) (6) (b) (6) plates, making it easy to thaw them and do the assays using a multichannel pipetor. Alternatively, we could store each serum in a tube, separately. Which do you prefer? Also, what volume of serum do you need from each bleed? I’d like to generate several aliquots/animal/bleed, and want to store them in convenient volumes. Let me know. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Wednesday, July 18, 2001 10:45 AM (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG (b) (6) Hi, (b) (6) I had planned to give you 10 ml (20 doses) of each vaccine. Would this be sufficient? I could give you more AVA if you like. I could also give you (individually) several hundred micrograms of PA, and several ml of PBS and Alhydrogel. Let me know and I’ll bring them to you when I bring the vaccines. Thanks. – Bruce —–Original Message—– Sent: Wednesday, July 18, 2001 10:33 AM To: ‘Ivins Bruce E’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, I don’t know how precious the two anthrax vaccines are. If you could spare additional material, I’d like to test it for the presence of DNA that might contain immunostimulatory or immunosuppressive sequences. We’d purify out the DNA and test it in vitro. This is part of an ongoing project in my lab where we test various vaccines for such agents. I’ll take anything you can spare. —–Original Message—– From: Ivins Bruce E [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, October 07, 1999 8:40 AM To: Subject: Anthrax, mice, and CpG Hi, As you remember, in our first experiment with the mice, we got some time-to-death extension with CpG for mice challenged with virulent B. anthracis spores. In the second experiment, we demonstrated not only time-to-death extension, but also protection from death with the CpG. In this last experiment which we just concluded, we strangely got no protection (b) (6) (b) (6) (b) (6) (b) (6) at all, in terms of either survival or increased time-to-death. I believe that the main problem is that the mouse is such a generally poor and unpredictable model for anthrax. The guinea pig is a MUCH better model for anthrax infection/protection, and our guinea pig protocol for CpG has been approved, so I think the next step should be (when we get the funds released) to go into the guinea pigs. We’ll be able to look at specific as well as non-specific protection, and if we get some promising results, we can head into non-human primates. Hopefully we’ll get some money released within a few weeks and we can get started then. I’ll let you know. I’m sure that mice are an excellent animal model for a number of diseases, but anthrax isn’t one of them. – Bruce From: To: Subject: Date: Hi, , (6) Ivins, Bruce E Dr USAMRIID “(b) (6) RE: spore work Monday, August 27, 2001 9:29:28 AM (b) If you have what we sent to you previously, GREAT! Go ahead and use that, since it’s what you used to prepare the last batch. Thanks, Give us a heads-up for when you plan to send the various shipments. (6()6) Have a good week! – Bruce —–Original Message—– From: Sent: Monday, August 27, 2001 9:26 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Cc: Subject: RE: spore work Hi Bruce, I do have an aliquot of AMES that you had sent previously if you’d like me to use that. I just thought you might have a specific stock from which you wanted me to start. If it’s all the same to you, then I’ll use what you had sent me for the last go round. Just let me know. Thanks, (6) —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Saturday, August 25, 2001 5:01 PM To: Subject: RE: spore work Hi, (6)That’s great news. If you want to initiate an EA101, then we’ll get the spores ready on this end. Let’s plan on sending them on Tuesday, September 4. That should give time to everyone on both ends. We’ll send you spores from the previous spores you sent us. They will be on gel ice. We’ll send you 1 ml, 3 X 10^10 per ml, in a Nunc polypropylene vial. If you need more, let me know. We’ll wait for the EA101. My FAX is who is in charge of safety here, has a FAX number of (6) . Our facility registration number is – Thanks again! – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 7:00 PM To: Ivins (E-mail) Cc: Subject: spore work (b()b) (b) (6) (b) (6) (b) (b) (6) (b) (b) (6) . (b) (b) (6) (b) (2) (b) (6) (b) (6) Bruce, I just received the test execution directive from the Program Analysis Office for the spore preparation. We’re good to go. Did you want to send a stock source of AMES for us to use? If you do, then I’ll initiate and EA 101 on Monday or whenever you want to do it. Regards and have a good weekend, (b) (6) From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID FW: rPA vaccine Monday, August 27, 2001 12:02:30 PM (b) (6) (b) (6) It sounds as if the FDA folks are desperate for the PA, since they already have ordered animals. I propose the following (which would be included in a CRDA): 1) that the PA will be shared with no other lab or other individuals in another lab. (I’m thinking specifically of 2) That any and all data generated with the PA be shared with us throughout and at the end of the experiment(s); 3) That the data not be published in abstract form, manuscript form, or shared with other individuals outside their lab without our prior approval. (Again this is to prevent them giving our data to for his benefit.) 4) That any material left over after the experiment be returned to us, and that no PA be used for work other than for the stated purpose of immunizing the 500 mice. If we present that to them and and they agree to it via email, we can go ahead and get a CRDA drawn up, have our meeting on September 3, and give them PA at that time. They would thus be committed to our terms prior to any transfer of PA. I’m not deliberately trying to screw somebody’s research, but I want the appropriate reassurances ahead of time, to be backed up by a CRDA. If you would prefer that they send a letter (official letterhead) agreeing to the above terms ahead of the CRDA, then that could work. It sounds as if they’re willing to do anything to get the PA. I think what I asked for in the first paragraph is reasonable. Please give me your opinion. I think if we get by email or official letterhead or FAX (or any combination of the 3) an agreement to abide by our conditions, we can give them the material as the CRDA is drawn up with the same conditions. – Bruce —–Original Message—– From: Sent: Monday, August 27, 2001 11:40 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: rPA vaccine Bruce, Things are getting complicated. I spoke with Biocon today, and was informed that the animals for the study have been purchased. That would be acceptable if we had a dosing schedule (we do) and if we had the antigen, as I assumed to be the case. Anyway, as you can perceive we (I) am in trouble now, and independently of the cause, I’d wish to get out of it. Certainly we appreciate all your help, and I believe that what you want to accomplish is reasonable. We are willing to sign any suitable agreement. If USAMRIID were a private enterprise, we would even offer to buy the material. However, this is a matter of timing now. I am willing to do what I can to resolve the issue. I absolutely can meet with you any time, for instance, on September 3. However, I would beg that you go to that meeting with options that ensure that the immunization of the mice is going to start on time. Please let me know the time of the meeting; I will try to get available by phone. Thanks. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Saturday, August 25, 2001 11:56 AM (b) (6) (b) (6) (b) (6) (b) (6) To: ‘ Subject: RE: rPA vaccine (6) I doubt that a CRDA can be signed by 15 SEP. The PA cost us approximately $1,000 per vial, and several vials will be needed for the mouse immunizations. believes that this should be a cooperative agreement, rather than an MTA, and that we need a meeting first. If you can meet with us at 10 am on September 3, I believe that we can straighten things out pretty much. I’m not sure how long a contract can be delayed there. Here at USAMRIID, we can extend payment past the FY until 31 DEcember. Can you meet here at 10 am on 3 September? – Bruce —–Original Message—– From: ] Sent: Friday, August 24, 2001 5:03 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: rPA vaccine Bruce, We set a contract with Biocon on the basis of the e-mail exchange below. Unfortunately, as I explained in a prior communication, won’t be back till September 7. I can probably ask the people of Biocon to delay the starting date of the project; however, I’m a little apprehensive about funding, due to end of the fiscal year issues. Do you think we can count on having the material within the first two weeks of September? Otherwise, I may be forced to cancel the contract, which I would prefer not to do. One more thing I can attempt to do is to arrange a conference call connecting from somewhere in the US. Do you want me to do that? Please explore (w6i)th such a possibility. Thanks. (6) (b) (b) (6) (b) (6) (b) (6) (b) (b) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: “(b) (6) Subject: RE: rPA vaccine Date: Monday, August 27, 2001 12:02:58 PM I’m working on it, . I’ll let you know as soon as possible. – Bruce (6) —–Original Message—– From: Sent: Monday, August 27, 2001 11:40 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: rPA vaccine Bruce, Things are getting complicated. I spoke with Biocon today, and was informed that the animals for the study have been purchased. That would be acceptable if we had a dosing schedule (we do) and if we had the antigen, as I assumed to be the case. There might have been some misunderstanding, but I interpreted your July 9 response as an indication that the antigen would be available for the 500 mice even in the absence of an MTA or CRDA. That is why I delayed also my review of the draft MTA. Anyway, as you can perceive we (I) am in trouble now, and independently of the cause, I’d wish to get out of it. Certainly we appreciate all your help, and I believe that what you want to accomplish is reasonable. We are willing to sign any suitable agreement. If USAMRIID were a private enterprise, we would even offer to buy the material. However, this is a matter of timing now. I am willing to do what I can to resolve the issue. I absolutely can meet with you any time, for instance, on September 3. However, I would beg that you go to that meeting with options that ensure that the immunization of the mice is going to start on time. Please let me know the time of the meeting; I will try to get available by phone. Thanks. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Saturday, August 25, 2001 11:56 AM (b) (b) (6) (b) (6) To: ‘ Subject: RE: rPA vaccine (6) I doubt that a CRDA can be signed by 15 SEP. The PA cost us approximately $1,000 per vial, and several vials will be needed for the mouse immunizations. believes that this should be a cooperative agreement, rather than an MTA, and that we need a meeting first. If you can meet with us at 10 am on September 3, I believe that we can straighten things out pretty much. I’m not sure how long a contract can be delayed there. Here at USAMRIID, we can extend payment past the FY until 31 DEcember. Can you meet here at 10 am on 3 September? – Bruce —–Original Message—– From: Sent: Friday, August 24, 2001 5:03 PM To: ‘Ivins, Bruce E Dr USAMRIID’ (b) (6) (b) (b) (6) (b) (6) Subject: RE: rPA vaccine Bruce, We set a contract with Biocon on the basis of the e-mail exchange below. Unfortunately, as I explained in a prior communication, won’t be back till September 7. I can probably ask the people of Biocon to delay the starting date of the project; however, I’m a little apprehensive about funding, due to end of the fiscal year issues. Do you think we can count on having the material within the first two weeks of September? Otherwise, I may be forced to cancel the contract, which I would prefer not to do. One more thing I can attempt to do is to arrange a conference call connecting Bruce from somewhere in the US. Do you want me to do that? Please explore with such a possibility. Thanks. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Monday, July 09, 2001 7:47 AM (b) (6) (b) (6) To: ‘ Ivins, Bruce E Dr USAMRIID Cc: Subject: RE: rPA vaccine Hi, (6)We will be happy to prepare more vaccine for you and have it ready for you on Jul 19. In addition, we can supply you with the vaccine for the 500 mice. The PA we have been using is the GLP/GMP PA produced at the MARP here at Fort Detrick. It is very high quality, but the supplies are becoming somewhat limited. Thus, if more PA vaccine is needed later, we may need to talk about your specific needs. (We have a great deal of PA which is not GLP/GMP, which is active in toxin neutralization tests, and which is protectively immunogenic. It’s just not the GLP/GMP material that we have used in the past.) Also, if more material is desired past that which is needed for the 500 mice, We should probably establish an MTA or a CRDA. Please let me know a little in advance exactly when you or your wife will be coming, so we can be sure to be here at that time. Thank you. – Regards, – Bruce —–Original Message—– From: Sent: Friday, July 06, 2001 3:23 PM To: ‘Bruce.Ivins@DET.AMEDD.ARMY.MIL’ Cc: Subject: rPA vaccine Dear Bruce, Thanks again for providing us with vaccine for our pilot study of rPA immunogenicity. We have gotten already some results that I’ll share with you as soon as they are in presentable shape. The next (third) immunization of the mice with rPA is scheduled for July 20. I’ll be out of town Monday through Wednesday of that week. Therefore, I want to ask you if either my wife or I can pick up the vaccine on Thursday, July 19. I remind you that in this instance groups of 10 mice will be immunized with 1, 3 and 9 micrograms per 0.5 ml dose, and that we will inject ten controls with 0.5 ml of adjuvant in saline. In addition, I want to explore the possibility of (b) (6) (b) (6) (b) (b) (6) (b) (6) enrolling your help in preparing the big batch of mouse anti-PA serum that is the final goal of this preliminary study. We are in the process of establishing a contract to get 500 mice immunized up to three times with a dose chosen on the basis of the results of the pilot study. I don’t anticipate the need of using the 18, 18, 9 micrograms schedule. Also we are not certain yet if a third dose will be necessary. Summarizing, do you think that USAMRIID (you) could provide us with up to 2 X 500 dose of vaccine (6 micrograms in 0.5 ml) plus 500 doses (3 micrograms in 0.5 ml). According to my calculations a total of 7.5 mg of rPA would be needed. As usual, a little excess would be necessary to ensure appropriate dosing. In addition of the acknowledgement of your contribution to the preparation of this material and access to our results, we are willing to share a fraction of the serum produced. I would like to hear your opinion about this project. We do not expect that the immunization of the 500 animals will begin before the conclusion of our pilot study, approximately on August 17. However, we should finish contract writing for the project as soon as possible, due to end of the fiscal year deadlines. Please let me know what do you think. Regards. From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID rPA Monday, August 27, 2001 1:01:21 PM (b) (6) (b) (b) (6) (6) I talked it over with and we think that we have a solution. We will start working on a CRDA here. Also, let’s plan on meeting here at 10 am on September 3, unless that’s bad for you. Below I will list what will go into the CRDA. If you will agree to the conditions by email, then send by regular mail, on official letterhead, a signed letter agreeing to them, then we can get the PA – tell us exactly how much you need, including extra – to you promptly. This is standard for what would be in a Cooperative Research and Development Agreement: 1) That we will provide you with the requested amount of purified recombinant protective antigen (rPA). lab. 2) That the rPA will not be shared or given to any other lab or any individuals in any other 3) That any and all data generated with the rPA experiments will be shared with us throughout and at the end of the experiment(s). 4) That the data will not be published, either in abstract or manuscript form, or shared with individuals outside your lab, without prior input and approval from us. 5) That any rPA left over after the experiments will be returned to us. 6) That the rPA will not be used for experiment other than the immunization of mice. I hope you are agreeable to the above. As soon as we hear from you both by email and regular U.S. mail, we will get to you the rPA. Although some of the conditions may seem a bit stringent, it is because we have had problems in the past with a) material we have sent to someone being given to other labs; b) data being published or shared with others without our knowledge or input; c) our material resulting in data that was never shared with us; d) that material we sent to individuals was used for other than stated purposes. If necessary, and if you want to hand carry a letter up here and hand carry the rPA back down with you, then we could get you the material this week. Sincerely, – Bruce Ivins From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID “(b) (6) RE: Anthrax, mice, and CpG Monday, August 27, 2001 3:20:14 PM Can I get this to you on Thursday or Friday? – Bruce —–Original Message—– From: Sent: Monday, August 27, 2001 2:06 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Thanks for the offer. We’re on the schedule to boost the animals on Tuesday, Sept 4. Thus, if you could get us 18 doses of each type of vaccine (AVA and rPA vaccine) plus alum sometime the week before, that would be super. All the best, —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Saturday, August 25, 2001 7:07 PM (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG I can get it to you this week, Please tell me the day you are planning to immunize. Also, how many doses of each (AVA and rPA vaccine) will you need. (Include any extra that you wish to have.) Thanks! – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 4:43 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Hope you are well. The monkeys were immunized and bled. We need the second dose of both types of vaccine for the booster immunization ASAP. We’ll re-bleed a few times, then send you the serum for testing. All the best, (b) (6) (b) (6) (b) (6) (b) (6) —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, July 19, 2001 6:27 PM To: ‘ Subject: RE: Anthrax, mice, and CpG Hi, , If you can get us 1 ml of serum, then we can probably go with that. – Bruce —–Original Message—– From: Sent: Thursday, July 19, 2001 5:41 PM To: Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, I spoke too soon. Given that we plan to bleed the animals every 2 weeks, the vets won’t allow us to draw 5 ml each time (to get 2 ml of serum). Especially since we also want to do some serum chemistry and hematology on these animals. Can you make do with less serum? I’m only guessing, but when we studied mouse serum for anti-PA Abs by ELISA, the assay only took a few microliters of serum. More serum was needed for the neutralization assays, but if memory serves, the total volume required was approximately 300 ul. Thus, if you could get by with 0.5 – 1 ml of serum, everything will be OK. Otherwise, let me know, and we’ll figure something out. —–Original Message—– From: Sent: Thursday, July 19, 2001 4:08 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Will do. At least two 1 ml tubes of serum will be frozen down from each animal at each time point. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, July 19, 2001 3:11 PM To: Subject: RE: Anthrax, mice, and CpG , We draw blood into serum separator tubes, draw off the serum after clotting, aliquot the serum as we wish, then freeze it. For small (about 1 ml or less) volumes, we use cryotubes. For larger volumes, we use 15-ml, screw-capped, conical polypropylene tubes. For bleed-outs (which I believe won’t be done in this experiment), we use 50-ml, screw-capped, conical polypropylene tubes. For each time-point in each monkey, we would like 2 (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) aliquots of 1 ml of serum (thus, 2 ml total). That is enough to cover both ELISA and TNA tests. After all the samples are taken, I can drive down and pick up the frozen serum, bring it back, and the ELISA and TNA assays will be run. Thanks. I’m eagerly looking forward to this experiment. – Bruce —–Original Message—– From: Sent: Thursday, July 19, 2001 1:57 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, We’re all set for the monkey study. The animals are scheduled to be immunized on Tuesday, the day after the vaccine arrives. If memory serves, we’re injecting 0.5 ml intramuscularly. They’ll be re-immunized one month later. Bloods will be drawn every two weeks, and serum stored for you to test. In terms of collecting and storing the serum, I could use your advice. We do many of our ELISA assays in microtiter plates. If the same is true at your end, I could store the serum samples from each bleed in microtiter plates, making it easy to thaw them and do the assays using a multichannel pipetor. Alternatively, we could store each serum in a tube, separately. Which do you prefer? Also, what volume of serum do you need from each bleed? I’d like to generate several aliquots/animal/bleed, and want to store them in convenient volumes. Let me know. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Wednesday, July 18, 2001 10:45 AM (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG Hi, I had planned to give you 10 ml (20 doses) of each vaccine. Would this be sufficient? I could give you more AVA if you like. I could also give you (individually) several hundred micrograms of PA, and several ml of PBS and Alhydrogel. Let me know and I’ll bring them to you when I bring the vaccines. Thanks. – Bruce —–Original Message—– From: Sent: Wednesday, July 18, 2001 10:33 AM To: ‘Ivins Bruce E’ Subject: RE: Anthrax, mice, and CpG (b) (6) (b) (6) (b) (6) Dear Bruce, I don’t know how precious the two anthrax vaccines are. If you could spare additional material, I’d like to test it for the presence of DNA that might contain immunostimulatory or immunosuppressive sequences. We’d purify out the DNA and test it in vitro. This is part of an ongoing project in my lab where we test various vaccines for such agents. I’ll take anything you can spare. —–Original Message—– From: Ivins Bruce E [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, October 07, 1999 8:40 AM To: Subject: Anthrax, mice, and CpG Hi, As you remember, in our first experiment with the mice, we got some time-to-death extension with CpG for mice challenged with virulent B. anthracis spores. In the second experiment, we demonstrated not only time-to-death extension, but also protection from death with the CpG. In this last experiment which we just concluded, we strangely got no protection at all, in terms of either survival or increased time-to-death. I believe that the main problem is that the mouse is such a generally poor and unpredictable model for anthrax. The guinea pig is a MUCH better model for anthrax infection/protection, and our guinea pig protocol for CpG has been approved, so I think the next step should be (when we get the funds released) to go into the guinea pigs. We’ll be able to look at specific as well as non-specific protection, and if we get some promising results, we can head into non-human primates. Hopefully we’ll get some money released within a few weeks and we can get started then. I’ll let you know. I’m sure that mice are an excellent animal model for a number of diseases, but anthrax isn’t one of them. – Bruce (b) (6) (b) (6) (b) (6) From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID RE: Anthrax, mice, and CpG Monday, August 27, 2001 7:37:48 PM (b) (6) (b) (6) Hi, Is just before noon on Thursday or Friday better for you? – Bruce —–Original Message—– From: Sent: Monday, August 27, 2001 5:00 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Absolutely. Let me know how we can arrange it. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Monday, August 27, 2001 3:20 PM (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG Can I get this to you on Thursday or Friday? – Bruce —–Original Message—– From: Sent: Monday, August 27, 2001 2:06 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Thanks for the offer. We’re on the schedule to boost the animals on Tuesday, Sept 4. Thus, if you could get us 18 doses of each type of vaccine (AVA and rPA vaccine) plus alum sometime the week before, that would be super. All the best, —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Saturday, August 25, 2001 7:07 PM (b) (6) (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG I can get it to you this week, Please tell me the day you are planning to immunize. Also, how many doses of each (AVA and rPA vaccine) will you need. (Include any extra that you wish to have.) Thanks! (b) (6) (b) (6) – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 4:43 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Hope you are well. The monkeys were immunized and bled. We need the second dose of both types of vaccine for the booster immunization ASAP. We’ll re-bleed a few times, then send you the serum for testing. All the best, —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, July 19, 2001 6:27 PM (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG Hi, If you can get us 1 ml of serum, then we can probably go with that. – Bruce —–Original Message—– From: Sent: Thursday, July 19, 2001 5:41 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, I spoke too soon. Given that we plan to bleed the animals every 2 weeks, the vets won’t allow us to draw 5 ml each time (to get 2 ml of serum). Especially since we also want to do some serum chemistry and hematology on these animals. Can you make do with less serum? I’m only guessing, but when we studied mouse serum for anti-PA Abs by ELISA, the assay only took a few microliters of serum. More serum was needed for the neutralization assays, but if memory serves, the total volume required was approximately 300 ul. Thus, if you could get by with 0.5 – 1 ml of serum, everything will be OK. Otherwise, let me know, and we’ll figure something out. —–Original Message—– From: Sent: Thursday, July 19, 2001 4:08 PM To: ‘Ivins, Bruce E Dr USAMRIID’ (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) Subject: RE: Anthrax, mice, and CpG Will do. At least two 1 ml tubes of serum will be frozen down from each animal at each time point. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, July 19, 2001 3:11 PM (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG We draw blood into serum separator tubes, draw off the serum after clotting, aliquot the serum as we wish, then freeze it. For small (about 1 ml or less) volumes, we use cryotubes. For larger volumes, we use 15-ml, screw-capped, conical polypropylene tubes. For bleed-outs (which I believe won’t be done in this experiment), we use 50-ml, screw-capped, conical polypropylene tubes. For each time-point in each monkey, we would like 2 aliquots of 1 ml of serum (thus, 2 ml total). That is enough to cover both ELISA and TNA tests. After all the samples are taken, I can drive down and pick up the frozen serum, bring it back, and the ELISA and TNA assays will be run. Thanks. I’m eagerly looking forward to this experiment. – Bruce —–Original Message—– From: Sent: Thursday, July 19, 2001 1:57 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, We’re all set for the monkey study. The animals are scheduled to be immunized on Tuesday, the day after the vaccine arrives. If memory serves, we’re injecting 0.5 ml intramuscularly. They’ll be re-immunized one month later. Bloods will be drawn every two weeks, and serum stored for you to test. In terms of collecting and storing the serum, I could use your advice. We do many of our ELISA assays in microtiter plates. If the same is true at your end, I could store the serum samples from each bleed in microtiter plates, making it easy to thaw them and do the assays using a multichannel pipetor. Alternatively, we could store each serum in a tube, separately. Which do you prefer? Also, what volume of serum do you need from each bleed? I’d like to generate several aliquots/animal/bleed, and want to store them in convenient volumes. Let me know. (b) (6) (b) (6) (b) (6) (b) (6) —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Wednesday, July 18, 2001 10:45 AM To: Subject: RE: Anthrax, mice, and CpG Hi, I had planned to give you 10 ml (20 doses) of each vaccine. Would this be sufficient? I could give you more AVA if you like. I could also give you (individually) several hundred micrograms of PA, and several ml of PBS and Alhydrogel. Let me know and I’ll bring them to you when I bring the vaccines. Thanks. – Bruce —–Original Message—– From: Sent: Wednesday, July 18, 2001 10:33 AM To: ‘Ivins Bruce E’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, I don’t know how precious the two anthrax vaccines are. If you could spare additional material, I’d like to test it for the presence of DNA that might contain immunostimulatory or immunosuppressive sequences. We’d purify out the DNA and test it in vitro. This is part of an ongoing project in my lab where we test various vaccines for such agents. I’ll take anything you can spare. —–Original Message—– From: Ivins Bruce E [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, October 07, 1999 8:40 AM To: Subject: Anthrax, mice, and CpG Hi, , As you remember, in our first experiment with the mice, we got some time-to-death extension with CpG for mice challenged with virulent B. anthracis spores. In the second experiment, we demonstrated not only time-to-death extension, but also protection from death with the CpG. In this last experiment which we just concluded, we strangely got no protection at all, in terms of either survival or increased time-to-death. I believe that the main problem is that the mouse is such a generally poor and unpredictable model for anthrax. The guinea pig is a MUCH better model for anthrax infection/protection, and our guinea pig protocol for CpG has been approved, so I think the next step should be (when we get the funds released) to go into the guinea pigs. We’ll be able to look at specific as well as non-specific protection, and if we get some promising results, we can head into non-human primates. Hopefully we’ll get some money released within a few weeks and we can get started then. I’ll let you know. I’m sure that mice are an excellent animal model for a number of diseases, but anthrax isn’t one of them. (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) – Bruce From: To: Subject: Date: Ivins, Bruce E Dr USAMRIID RE: Anthrax, mice, and CpG Tuesday, August 28, 2001 8:41:45 AM (b) (6) (b) (6) Friday would be better for me, If you like, you could mail frozen samples to me, I could pick them up, or you could bring them up. Whichever you prefer is fine. Also, the American Society for Microbiology Annual Meeting will be next May, and I would like to present an abstract (with the two of us as the authors) with our guinea pig and (hopefully) monkey data. I would send the abstract to you for your comments, criticisms, suggestions, additions, deletions, etc., before submitting it. Would this be acceptable to you? I think it would be very well received at the meeting. I’ll see you Friday, a little before noon, if it’s convenient. – Bruce —–Original Message—– From: Sent: Tuesday, August 28, 2001 8:34 AM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Either one is fine. Just let me know what’s best. Since I feel bad about you having to run down here so often, how about I agree to bring the serum samples up to you when they’re all collected? —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Monday, August 27, 2001 7:38 PM (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG Hi, , Is just before noon on Thursday or Friday better for you? – Bruce —–Original Message—– From: Sent: Monday, August 27, 2001 5:00 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Absolutely. Let me know how we can arrange it. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Monday, August 27, 2001 3:20 PM (b) (6) (b) (6) (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG Can I get this to you on Thursday or Friday? – Bruce —–Original Message—– From: Sent: Monday, August 27, 2001 2:06 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Thanks for the offer. We’re on the schedule to boost the animals on Tuesday, Sept 4. Thus, if you could get us 18 doses of each type of vaccine (AVA and rPA vaccine) plus alum sometime the week before, that would be super. All the best, —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Saturday, August 25, 2001 7:07 PM To: Subject: RE: Anthrax, mice, and CpG I can get it to you this week, Please tell me the day you are planning to immunize. Also, how many doses of each (AVA and rPA vaccine) will you need. (Include any extra that you wish to have.) Thanks! – Bruce —–Original Message—– From: Sent: Thursday, August 23, 2001 4:43 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, Hope you are well. The monkeys were immunized and bled. We need the second dose of both types of vaccine for the booster immunization ASAP. We’ll re-bleed a few times, then send you the serum for testing. All the best, —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Thursday, July 19, 2001 6:27 PM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) To: ‘ Subject: RE: Anthrax, mice, and CpG Hi, If you can get us 1 ml of serum, then we can probably go with that. – Bruce —–Original Message—– From: Sent: Thursday, July 19, 2001 5:41 PM To: ‘Ivins, Bruce E Dr USAMRIID’ Subject: RE: Anthrax, mice, and CpG Dear Bruce, I spoke too soon. Given that we plan to bleed the animals every 2 weeks, the vets won’t allow us to draw 5 ml each time (to get 2 ml of serum). Especially since we also want to do some serum chemistry and hematology on these animals. Can you mak
        • DXer said

          In a May 2001 email, in an email titled “Long-term efficacy study in rabbits and monkeys,” Dr. Ivins wrote:

          “Should part of the studies be contracted out in order to save USAMRIID animal room space? Where? SRI? Covance? SAIC?”

          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: Long-term efficacy study in rabbits and monkeys
          Date: Tuesday, May 08, 2001 4:03:03 PM
          Hi,
          Here are some ideas based on what we talked about yesterday with respect to a long-term efficacy study in rabbits and monkeys:
          1. Vaccine: NEW recombinant PA (50 micrograms) + Dulbecco’s PBS + Alhydrogel (0.5 mg metallic aluminum) in a total volume of 0.5 ml, with or without formaldehyde (to be determined prior to experiment)
          2. Animals – New Zealand white rabbits, 2.5-3.5 kg and rhesus macaques, adults.
          3. Challenge – aerosol challenge with 100 – 500 LD50 of B. anthracis Ames spores.
          4. Immunization and challenge schedule for rabbits (10 males and 10 females per challenge group & 2 males and 2 females per control group):
          0 month and 1 month (4 weeks) – immunize 160 rabbits (8 groups of 20) with rPA vaccine, and immunize 8 control groups of rabbits with PBS
          3 months – challenge group 1 and group 1 controls
          6 months – challenge group 2 and group 2 controls
          12 months – challenge group 3 and group 3 controls
          12 months – immunize group 6 rabbits (with rPA vaccine) and group 6 controls (with PBS); if group 3 challenged animals have less than 80% (?) survival, challenge group 6 and group 6 control rabbits at 13 months
          18 months – challenge group 4 rabbits and group 4 controls
          18 months – immunize group 7 rabbits (with rPA vaccine) and group 7 controls (with PBS); if group 4 challenged animals have less than 80% (?) survival, challenge group 7 and group 7 control rabbits at 19 months
          24 months – challenge group 5 rabbits and group 5 controls
          24 months – immunize group 8 rabbits (with rPA vaccine) and group 8 controls (with PBS); if group 5 challenged animals have less than 80% (?) survival, challenge group 8 and group 8 control rabbits at 25 months
          5. Bleeding schedule for rabbits:
          1 week prior to 3 months – all rabbits
          4, 7, 10 and 14 days after challenge – group 1
          1 week prior to 6 months – all remaining rabbits
          4, 7, 10 and 14 days after challenge – group 2
          1 week prior to 12 months – all remaining rabbits
          4, 7, 10 and 14 days after challenge – group 3
          1 week prior to 13 months – group 6 and group 6 controls
          4, 7, 10 and 14 days after challenge – group 6
          1 week prior to 18 months – all remaining rabbits
          4, 7, 10 and 14 days after challenge – group 4
          1 week prior to 19 months – group 7 and group 7 controls
          4, 7, 10 and 14 days after challenge – group 7
          1 week prior to 24 months – all remaining rabbits
          4, 7, 10 and 14 days after challenge – group 5
          1 week prior to 25 months – group 8 and group 8 controls
          4, 7, 10 and 14 days after challenge – group 8
          6. Blood and sera will be assayed by ELISA, TNA, & ELISPOT.
          *************************************************************************************************************************************************
          7. Immunization and challenge schedule for monkeys (5 males and 5 females per challenge group & 1 male and 1 female per control group):
          0 month and 1 month (4 weeks) – immunize 77 monkeys (7 groups of 10) with rPA vaccine, and immunize 7 control groups with PBS
          6 months – challenge group 1 and group 1 controls
          12 months – challenge group 2 and group 2 controls
          12 months – immunize group 5 monkeys (with rPA vaccine) and group 5 controls (with PBS); if group 3 challenged animals have less than 80% (?) survival, challenge group 5 and group 5 control monkeys at 13
          months
          24 months – challenge group 3 rabbits and group 3 controls
          24 months – immunize group 6 monkeys (with rPA vaccine) and group 6 controls (with PBS); if group 5 challenged animals have less than 80% (?) survival, challenge group 6 and group 6 control monkeys at 25
          months
          36 months – challenge group 4 monkeys and group 4 controls
          36 months – immunize group 7 monkeys (with rPA vaccine) and group 7 controls (with PBS); if group 7 challenged animals have less than 80% (?) survival, challenge group 7 and group 7 control monkeys at 37
          months
          8. Bleeding schedule for rabbits:
          1 week prior to 6 months – all monkeys
          4, 7, 10 and 14 days after challenge – group 1
          1 week prior to 12 months – all remaining rabbits
          4, 7, 10 and 14 days after challenge – group 2
          1 week prior to 13 months – group 5 and group 5 controls
          4, 7, 10 and 14 days after challenge – group 5
          1 week prior to 24 months – all remaining rabbits
          4, 7, 10 and 14 days after challenge – group 3
          1 week prior to 25 months – group 6 and group 6 controls
          4, 7, 10 and 14 days after challenge – group 6
          1 week prior to 36 months – all remaining rabbits
          4, 7, 10 and 14 days after challenge – group 4
          1 week prior to 37 months – group 7 and group 7 controls
          4, 7, 10 and 14 days after challenge – group 7
          9. Blood and sera will be assayed by ELISA, TNA, & ELISPOT.
          ***************************************************************************************************************************************************************************************
          10. Surviving rabbits will be euthanized. Surviving monkeys will be “grayed out.”
          11. Rabbits will be challenged at 13, 19 or 25 months only if there is less than 80% survival at 12, 18 or 24 months.
          12. Monkeys will be challenged at 13, 25 or 37 months, only if there is less than 80% survival at 12, 24 or 36 months.
          ***********************************************************************************************************************************************************************************
          13, Points to consider:
          a) Should part of the studies be contracted out in order to save USAMRIID animal room space? Where? SRI? Covance? SAIC?
          b) Other points?

        • DXer said

          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: RE: Anthrax, mice, and CpG
          Date: Friday, August 31, 2001 8:47:03 AM
          I plan to leave about 10 am, so I’ll be there about 11 am. Could I get another parking exemption while
          I’m there? Also, we are running short of the FAV038 lot of AVA, so it would be greatly
          appreciated if we could get back any left over AVA after the experiment. Thanks! We are bringing you
          today:
          1) AVA (2 vials, 10 ml total)
          2) PA/Alhydrogel/PBS (10 ml total)
          3) Control – Alhydrogel/PBS (no PA) (10 ml total)
          If this is not OK, please let me know before I leave. Thanks!
          – Bruce
          —–Original Message—–
          From:
          Sent: Friday, August 31, 2001 8:30 AM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          Bruce,
          We’ve cancelled lab meeting this morning, so I’m available at any time for
          the anthrax vaccine. Give a buzz before you leave to drop it off, so I can
          make sure I’m waiting for you.
          All the best,
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Tuesday, August 28, 2001 8:42 AM
          To: ‘
          Subject: RE: Anthrax, mice, and CpG
          Friday would be better for me, If you like, you could mail
          frozen samples to me, I could pick them up, or you could bring them up.
          Whichever you prefer is fine. Also, the American Society for Microbiology
          Annual Meeting will be next May, and I would like to present an abstract
          (with the two of us as the authors) with our guinea pig and (hopefully)
          monkey data. I would send the abstract to you for your comments, criticisms,
          suggestions, additions, deletions, etc., before submitting it. Would this be
          acceptable to you? I think it would be very well received at the meeting.
          I’ll see you Friday, a little before noon, if it’s convenient.
          – Bruce
          —–Original Message—–
          From:
          Sent: Tuesday, August 28, 2001 8:34 AM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          (b) (6)
          (b) (6)
          (b) (6)
          (b) (6)
          (b) (6)
          (b) (6)
          (b) (6)
          Dear Bruce,
          Either one is fine. Just let me know what’s best.
          Since I feel bad about you having to run down here so often, how about I
          agree to bring the serum samples up to you when they’re all collected?
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Monday, August 27, 2001 7:38 PM
          To: ‘
          Subject: RE: Anthrax, mice, and CpG
          Hi,
          Is just before noon on Thursday or Friday better for you?
          – Bruce
          —–Original Message—–
          From:
          Sent: Monday, August 27, 2001 5:00 PM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          Absolutely. Let me know how we can arrange it.
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Monday, August 27, 2001 3:20 PM
          To: ‘
          Subject: RE: Anthrax, mice, and CpG
          Can I get this to you on Thursday or Friday?
          – Bruce
          —–Original Message—–
          From:
          Sent: Monday, August 27, 2001 2:06 PM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          Dear Bruce,
          Thanks for the offer. We’re on the schedule to boost the animals on
          Tuesday, Sept 4. Thus, if you could get us 18 doses of each type of vaccine
          (AVA and rPA vaccine) plus alum sometime the week before, that would be
          super.
          All the best,
          (b) (6)
          (b) (6)
          (b) (6)
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          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Saturday, August 25, 2001 7:07 PM
          To:
          Subject: RE: Anthrax, mice, and CpG
          I can get it to you this week, Please tell me the day you are
          planning to immunize. Also, how many doses of each (AVA and rPA vaccine)
          will you need. (Include any extra that you wish to have.) Thanks!
          – Bruce
          —–Original Message—–
          From:
          Sent: Thursday, August 23, 2001 4:43 PM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          Dear Bruce,
          Hope you are well.
          The monkeys were immunized and bled. We need the second dose of both types
          of vaccine for the booster immunization ASAP. We’ll re-bleed a few times,
          then send you the serum for testing.
          All the best,
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Thursday, July 19, 2001 6:27 PM
          Subject: RE: Anthrax, mice, and CpG
          Hi, ,
          If you can get us 1 ml of serum, then we can probably go with that.
          – Bruce
          —–Original Message—–
          From:
          Sent: Thursday, July 19, 2001 5:41 PM
          To: Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          Dear Bruce,
          I spoke too soon. Given that we plan to bleed the animals every 2 weeks,
          the vets won’t allow us to draw 5 ml each time (to get 2 ml of serum).
          Especially since we also want to do some serum chemistry and hematology on
          these animals.
          Can you make do with less serum? I’m only guessing, but when we studied
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          mouse serum for anti-PA Abs by ELISA, the assay only took a few microliters
          of serum. More serum was needed for the neutralization assays, but if
          memory serves, the total volume required was approximately 300 ul. Thus, if
          you could get by with 0.5 – 1 ml of serum, everything will be OK.
          Otherwise, let me know, and we’ll figure something out.
          —–Original Message—–
          From:
          Sent: Thursday, July 19, 2001 4:08 PM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          Will do. At least two 1 ml tubes of serum will be frozen down from each
          animal at each time point.
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Thursday, July 19, 2001 3:11 PM
          To:
          Subject: RE: Anthrax, mice, and CpG
          We draw blood into serum separator tubes, draw off the serum after
          clotting, aliquot the serum as we wish, then freeze it. For small (about 1
          ml or less) volumes, we use cryotubes. For larger volumes, we use 15-ml,
          screw-capped, conical polypropylene tubes. For bleed-outs (which I believe
          won’t be done in this experiment), we use 50-ml, screw-capped, conical
          polypropylene tubes. For each time-point in each monkey, we would like 2
          aliquots of 1 ml of serum (thus, 2 ml total). That is enough to cover both
          ELISA and TNA tests. After all the samples are taken, I can drive down and
          pick up the frozen serum, bring it back, and the ELISA and TNA assays will
          be run. Thanks. I’m eagerly looking forward to this experiment.
          – Bruce
          —–Original Message—–
          From:
          Sent: Thursday, July 19, 2001 1:57 PM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Anthrax, mice, and CpG
          Dear Bruce,
          We’re all set for the monkey study. The animals are scheduled to be
          immunized on Tuesday, the day after the vaccine arrives. If memory serves,
          we’re injecting 0.5 ml intramuscularly. They’ll be re-immunized one month
          later. Bloods will be drawn every two weeks, and serum stored for you to
          test.
          In terms of collecting and storing the serum, I could use your advice. We
          do many of our ELISA assays in microtiter plates. If the same is true at
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          your end, I could store the serum samples from each bleed in microtiter
          plates, making it easy to thaw them and do the assays using a multichannel
          pipetor. Alternatively, we could store each serum in a tube, separately.
          Which do you prefer? Also, what volume of serum do you need from each
          bleed? I’d like to generate several aliquots/animal/bleed, and want to
          store them in convenient volumes.
          Let me know.
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Wednesday, July 18, 2001 10:45 AM
          To: ‘
          Subject: RE: Anthrax, mice, and CpG
          Hi,
          I had planned to give you 10 ml (20 doses) of each vaccine. Would
          this be sufficient? I could give you more AVA if you like. I could also give
          you (individually) several hundred micrograms of PA, and several ml of PBS
          and Alhydrogel. Let me know and I’ll bring them to you when I bring the
          vaccines. Thanks.
          – Bruce
          —–Original Message—–
          From:
          Sent: Wednesday, July 18, 2001 10:33 AM
          To: ‘Ivins Bruce E’
          Subject: RE: Anthrax, mice, and CpG
          Dear Bruce,
          I don’t know how precious the two anthrax vaccines are. If you could spare
          additional material, I’d like to test it for the presence of DNA that might
          contain immunostimulatory or immunosuppressive sequences. We’d purify out
          the DNA and test it in vitro. This is part of an ongoing project in my lab
          where we test various vaccines for such agents.
          I’ll take anything you can spare.
          —–Original Message—–
          From: Ivins Bruce E [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Thursday, October 07, 1999 8:40 AM
          To: ‘
          Subject: Anthrax, mice, and CpG
          Hi, ,
          As you remember, in our first experiment with the mice, we got some
          time-to-death extension with CpG for mice challenged with virulent B.
          anthracis spores. In the second experiment, we demonstrated not only
          time-to-death extension, but also protection from death with the CpG. In
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          this last experiment which we just concluded, we strangely got no protection
          at all, in terms of either survival or increased time-to-death. I believe
          that the main problem is that the mouse is such a generally poor and
          unpredictable model for anthrax. The guinea pig is a MUCH better model for
          anthrax infection/protection, and our guinea pig protocol for CpG has been
          approved, so I think the next step should be (when we get the funds
          released) to go into the guinea pigs. We’ll be able to look at specific as
          well as non-specific protection, and if we get some promising results, we
          can head into non-human primates. Hopefully we’ll get some money released
          within a few weeks and we can get started then. I’ll let you know. I’m sure
          that mice are an excellent animal model for a number of diseases, but
          anthrax isn’t one of them.
          – Bruce
          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: NGAV meeting at Pentagon, 23 AUG 01
          Date: Friday, August 31, 2001 3:45:43 PM
          Here are some notes from the NGAV meeting at the Pentagon on 23 AUG:
          1) – For NGAV funding, FY01 funds are going to be freed up for work on VEE. This will allow
          some money to be made available for rPA funds.
          2) – BDRP (nee SAIC; nee MARP) has made some untenable demands with respect to the rPA
          made for human use. They want $200,000 per year (without specifying the number of years) for
          indemnification. They want the government to cover the cost of lawsuits. They also want international
          indemnification. They are refusing to release the rPA unless the Army agrees to the above demands.
          said that if the rPA is not given to the Army, then we may have to go to AVANT and use E.
          coli rPA. It was pointed out that since the E. coli rPA differs in some respects from the B. anthracis rPA,
          a number of studies will probably have to be repeated, including stability and efficacy studies. This will
          extend the timeline for rPA going into humans. said that this issue “is a RIID issue.” Several
          individuals disagreed with him on this statement. told o have this
          issue resolved by the next meeting.
          3) A new GANTT chart will be constructed, in color, to show what has been done and what needs to be
          done.
          4) said that NIH people are doing a lot of tech base studies on rPA, including the need
          for an adjuvant, passive immunity studies, surrogate marker studies, and stability/potency testing. Some
          of these studies are being conducted with CDC and include AVA studies.
          5) asked for a RIID tech base update at the next meeting on Friday, September
          28, 9-10:30 am, in office. This update should cover timelines, start, end, and cost
          for: 1) formulation studies; 2) stability studies; 3) passive immunity studies (here and at Battelle); 4)
          surrogate marker and efficacy studies; 5) CpG studies. ( – I don’t think that wants reams of
          information, just some bullets about what we’ve done and what is to be done, also, timelines and
          funding for the projects.)
          6) Action Items for next meeting :
          1) will have resolved the rPA issue.
          2) A new GANTT chart will have been made.
          3) will discuss studies of AVA in humans.
          4) will discuss tech base studies including Battelle and RIID studies.
          – Bruce
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: RE: Protocol modifications
          Date: Tuesday, September 04, 2001 2:01:53 PM
          Ninety-four rabbits won’t be a problem as far as the sub cu challenge goes, but will you have enough of
          each isoform to do the immunizations?
          – Bruce
          >—–Original Message—–
          >From:
          >Sent: Tuesday, September 04, 2001 8:36 AM
          >To:
          vins, Bruce E Dr USAMRIID
          >Subject: FW: Protocol modifications
          >
          >Re: animal protocol to study the contribution of rPA isoforms to protection against a lethal anthrax
          infection in rabbits. suggests we do 30 rabbits per group; see below. The change would
          result in 94 rabbits instead of 52 ($$$?). Please advise if we wish to accept his recommendations so I
          can make the changes. ( had originally suggested 16 per group). Thanks in advance.
          File: Rabbit isoform protocol23Aug.doc >>
          >>
          —–Original Message—–
          >From:
          >Sent: Friday, August 31, 2001 3:30 PM
          >To:
          >Subject: Protocol modifications
          >>>
          The isoform protocol is suggested to undergo the following statistical revisions. There are three
          reasons
          > for the revisions:
          >>
          1. This study is approaching the final formulation of the rPA anthrax vaccine candidate and thus
          must
          > adhere to stricter criteria.
          >>
          2. The stricter criteria involve the concept of “equivalency” testing. This means that we seek to
          reject
          > the hypothesis that the two isoforms are NOT equivalent with respect to survival and immune
          response
          > as measured by ELISA and TNA. Rejection of the hypothesis establishes the statistical evidence
          that the
          > isoforms are equivalent. This requires larger sample sizes than a test of the hypothesis that the
          two
          > isoforms are not different (a subtle difference), which is an early developmental hypothesis.
          >>
          3. Should this not be a strict GLP study due to the need for FDA approval of the decision that will
          > arise from it? We do not wish to have to repeat the study in the future under GLP standards.
          >>
          Revisions suggested:
          >>
          Page 3 replace objective with: “The hypothesis is that the two isoforms are NOT equivalent in
          survival rates
          > or immune response as measured by either ELISA and TNA. Equivalency is defined as a
          difference
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          > in survival rates of no more than 20%, and a ratio of geometric mean ELISA titers and TNA
          titers of
          > no more than 4-fold respectively. The hypothesis will be tested at the 95% confidence level
          (2-tailed). Rejection
          > of the hypothesis establishes the equivalency of the survival and immune response of the
          two isoforms.
          > Failure to reject any of the hypothesis (survival or immune response) fails to establish the
          equivalency
          > of the two isoforms. Sex-specific tests will be done.”
          >>
          Page 11 replace Data Analysis with: “Sex-specific survival rates will be tested for equivalence using
          the method
          > of Farrington and Manning (ref). ELISA and TNA titers will be transformed to log10 and
          analyzed for
          > sex-specific equivalency based on the 95% confidence intervals (2-tailed) around the ratio of
          geometric mean titers.
          > Further analysis of survival correlations with sex, titers and isoform will use standard logistic
          regression.
          > Also, further analysis of titers will use multiple regression to test for sex differences and to
          test for
          > isoform differences. All tests will be at the 95% confidence level (2-tailed). All data will be
          automated
          > and verified prior to analysis. Statistical software package SAS (version 8.0 or greater) will
          be used
          > for analysis.”
          >>
          Page 11 Move the sample size justification to the page 5 and replace the statement there with:
          >>
          The test of equivalency of survival rates assumes 100% survival in both isoforms and a
          20%
          > maximum difference resulting in a sample size of 15 of each sex per isoform. The test of
          > equivalency of immune response assumes a log10 standard deviation of 0.50 logs and a
          > maximum ratio of geometric mean titers of 4-fold (0.60 logs) resulting in a sample
          > size of 12 of each sex per isoform. Therefore, 30 rabbits (15 male and 15 female) must be
          > tested at each isoform and with the combined isoforms as a control. The unprotected
          controls
          > will be set at a nominal 4 animals (2 of each sex) due to confidence in the 100% lethality
          > of the challenge dose.”
          >>
          Reference: Farrington CP and Manning G. Test statistics and sample size formulae for comparative
          > binomial trials with null hypothesis of non-zero risk difference or non-unity relative risk.
          > Statistics in Medicine, vol 9, 147-1454, 1990.
          >
          >
          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: Preparation of rPA vaccine + Alhydrogel
          Date: Wednesday, September 05, 2001 8:07:57 AM
          Dear ,
          Here is some of the information you requested. I am happy to provide it to you.
          Your fiend,
          Bruce
          >Here is how we prepare our rPA vaccine with aluminum hydroxide adjuvant:
          >
          >Materials:
          >>
          1) Dulbecco’s phosphate buffered saline (PBS) without calcium and without magnesium, pH 7.4 –
          sterile
          > 2) Aluminum hydroxide (Alhydrogel) (2% Aluminum oxide) – sterilized by autoclaving
          > 3) PA in ammonim acetate buffer, pH 8.9
          >
          >The PA we have been using has a concentration of 1.18 mg/ml. (This can be measured by absorbance
          at 280 nm, since 1.0 mg/ml of PA gives a reading of 1.0.) We dilute the commercial Alhydrogel stock
          above 1:10.64 (final dilution in vaccine) to get 0.5 mg of metallic aluminum per 0.5 ml dose of vaccine.
          (I believe that the maximum allowed is 0.85 mg per dose. The current human anthrax vaccine has
          about 0.725 mg aluminum per dose. Below is how we would make up ten 0.5-ml doses of vaccine
          containing 50 micrograms of PA per dose and 0.5 mg of aluminum per dose:
          >>>
          0.424 ml of PA (500 micrograms of the 1.18 mg/ml material)
          > 0.470 ml of Alhydrogel
          > 4.106 ml of PBS
          >
          >Add the Alhydrogel to the PBS in a sterile vaccine vial and mix by swirling until an even suspension is
          achieved. Add the PA and swirl immediately to mix. Put on ice and allow to stand from 1 hour to
          overnight on ice. If desired, a small, sterile stirring bar can be added to the PBS + Alhydrogel. While on
          a magnetic mixer, add the PA. (We’ve never had a problem doing it the former way, without a stirring
          bar.)
          >
          >Hope this has been helpful.
          >>
          – Bruce
          >
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: FW: August NGAV Meeting Moved Again to Wednesday, August 29th, 11 00-1 200 hrs
          Date: Wednesday, September 05, 2001 8:20:18 AM
          Attachments:
          Here you go,
          – Bruce
          —–Original Message—–
          From:
          Sent: Monday, August 27, 2001 10:39 AM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: August NGAV Meeting Moved Again to Wednesday, August 29th,
          11 00-1 200 hrs
          Bruce,
          The Read Ahead is provided to only. The minutes from July are
          attached.
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Monday, August 27, 2001 10:38 AM
          To:
          Subject: FW: August NGAV Meeting Moved Again to Wednesday, August 29th,
          11 00-1 200 hrs
          Can you send to me the attachments? I never got them. Thanks! I’ll
          see you on Wednesday morning, about 10:30 am.
          – Bruce
          > —–Original Message—–
          > From: Ivins, Bruce E Dr USAMRIID
          > Sent: Monday, August 27, 2001 8:54 AM
          > To:
          > Subject: RE: August NGAV Meeting Moved Again to Wednesday, August
          29th, 1100-1 200 hrs
          >>
          – Do you have the NGAV Final Minutes-23 July.doc and
          NGAV Read Ahead-Aug? Please forward them to me if you have them so that I
          can read them before I go to the Pentabon on Wednesday.
          > Thanks.
          > – Bruce
          >>
          —–Original Message—–
          > From:
          > Sent: Wednesday, August 01, 2001 12:25 PM
          > To:
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          >
          > Subject: Approved Minutes for 23 July NGAV Meeting
          >>>
          Ladies and Gentlemen,
          >>
          Attached are the approved Minutes from the 23 July NGAV
          meeting. Please
          > note the new Action Items and the POC responsible to address
          them. I have
          > also attached a PowerPoint template to facilitate your
          response for the 30
          > August Meeting. The suspense for input to the Read Ahead
          for
          > will be NLT noon, Monday, 27 August. NOTE: the date of the
          August NGAV
          > meeting is 30 August, 0900-1030 hrs, Pentagon, Room 3C257.
          It does not
          > start at 0930 as previously mentioned at the last meeting.
          >>
          <> <>
          >>>
          >>
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: FW: Covance contract information
          Date: Wednesday, September 05, 2001 9:13:24 AM
          >—–Original Message—–
          >From:
          >Sent: Wednesday, September 05, 2001 8:17 AM
          >To: Ivins, Bruce E Dr USAMRIID
          >Subject: Covance contract information
          >>>>
          >Let me know when you plan to go and hopefully I can join you one day
          >
          >Thanks,
          >
          >>>
          >This contract is for the vaccination of 100 NZW rabbits on days 0 and 28. Includes prebleeds, im
          injections, terminal bleeds, maintenance, GLP study procedures, and cost of transportation and
          packaging.
          >
          >Pertinent Alores information for this contract is as follows:
          >Document No. W23MYC1159N015
          >APC 6R56
          >HR R8R
          >DAC
          >Purchase Order No. 01P0831
          >
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          From: Ivins, Bruce E Dr USAMRIID
          To: Ivins, Bruce E Dr USAMRIID;
          Subject: RE: Covance contract information
          Date: Wednesday, September 05, 2001 9:26:51 AM
          Addendum – in general, the best days for us are Mondays, Tuesdays and Thursdays. Wednesdays are
          usually full of animal work, and Fridays are frequently days of leave (use or lose). Other bad days –
          October 2, October 8, November 12, week of November 26.
          – Bruce
          >—–Original Message—–
          >From: Ivins, Bruce E Dr USAMRIID
          >Sent: Wednesday, September 05, 2001 8:34 AM
          >To:
          >Subject: RE: Covance contract information
          >
          >Dear ,
          >Let us know what days are especially bad for you (you’ll be on leave, at some wonderful meeting,
          etc.) or especially good for you. We’ll match that up with what days are good and bad for us and come
          up with a time to go. Thanks!!
          >- Brucie
          >>
          —–Original Message—–
          > From:
          > Sent: Wednesday, September 05, 2001 8:17 AM
          > To: Ivins, Bruce E Dr USAMRIID
          > Subject: Covance contract information
          >>>>>
          Let me know when you plan to go and hopefully I can join you one day
          >>
          Thanks,
          >>>>>>
          This contract is for the vaccination of 100 NZW rabbits on days 0 and 28. Includes prebleeds, im
          injections, terminal bleeds, maintenance, GLP study procedures, and cost of transportation and
          packaging.
          >>
          Pertinent Alores information for this contract is as follows:
          > Document No. W23MYC1159N015
          > APC 6R56
          > HR R8R
          > DAC
          > Purchase Order No. 01P0831
          >
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: FW: ABCNEWS.com: Researcher Infected With Bio-Warfare Agent
          Date: Wednesday, September 05, 2001 9:49:10 AM
          I thought you might find this story interesting – Bruce
          Researcher Infected With Bio-Warfare Agent
          http://abcnews.go.com/sections/living/DailyNews/biowarfare010725.html
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject:
          Date: Wednesday, September 05, 2001 4:15:59 PM
          Hi,
          has just told me that she is beginning to think of career advancement, and she is starting to
          look at other openings in the research technician field, including positions at USAMRIID. We’ve told her
          that this position is finite in duration, although we haven’t told her that this will be her last year in the
          position, beacause the position will end this coming July. We will inform her about this in January. In
          the meantime, please keep your eye open for positions around here that she may be interested in, and
          let her know of any. If she finds one that requires her to leave before July, it will be OK, as long as she
          gives us some lead time on leaving. She has had some very good training with us, and she can enter
          the biocontainment suites, and that is a very big plus at USAMRIID.
          Thanks!
          – Bruce
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: Ames spores
          Date: Thursday, September 06, 2001 9:42:11 AM
          Hi,
          I talked to in contracts here, and he said that everything is ready to go as far as
          the contract for the production of the Ames spores. We’re looking forward to getting them – we’re down
          to about 1/3 of the spore prep that you made previously – and then purifying them. Please let us know
          ahead of time when we should start to expect them.
          Thanks!! Hope you have a good fall!
          – Bruce
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: FW: Ames spores
          Date: Thursday, September 06, 2001 10:17:03 AM
          Info on the Dugway Ames spores. – Bruce
          —–Original Message—–
          From:
          Sent: Thursday, September 06, 2001 10:28 AM
          To: ‘Ivins, Bruce E Dr USAMRIID’
          Subject: RE: Ames spores
          Hi Bruce,
          I’m getting ready to have some maintenance performed on the
          fermentor that we will use. We’ll probably start up in about a month. I’ve
          got a couple of other runs to do first. I’m having a wonderful introduction
          to Fall with my football team at 2 and 0 and the temperature starting to
          drop.
          —–Original Message—–
          From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
          Sent: Thursday, September 06, 2001 7:42 AM
          To: ‘
          Subject: Ames spores
          Hi,
          I talked to in contracts here, and he said that
          everything is ready to go as far as the contract for the production of the
          Ames spores. We’re looking forward to getting them – we’re down to about 1/3
          of the spore prep that you made previously – and then purifying them. Please
          let us know ahead of time when we should start to expect them.
          Thanks!! Hope you have a good fall!
          – Bruce
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: Rabbit Immunizations
          Date: Monday, September 10, 2001 10:00:49 AM
          said that she has spoken to you about immunizing 100 more rabbits with recombinant
          B. anthracis protective antigen (with Alhydrogel adjuvant), 0.5 ml per intramuscular dose. We would like
          to bring the vaccine to you on Tuesday, September 18, and then 4 weeks later on Tuesday, October 16.
          We will give you 120 doses (60 ml total) to allow for a little extra.
          If these are not acceptable dates, please let me know. Thank you.
          – Bruce Ivins
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: Anti-PA Human serum
          Date: Monday, September 10, 2001 12:43:05 PM
          Hi,
          We have Anti-PA IgG from humans which we are using in the mouse passive protection studies. Is
          there any human anti-PA serum (not human IgG) which we could use in these studies? We would need
          about 100 ml for each experiment.
          Thanks!
          – Bruce
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: CpG progress update
          Date: Monday, September 10, 2001 1:23:05 PM
          Here is a progress update on CpG studies in animal hosts:
          1) In guinea pigs there is no non-specific protection afforded by CpG oligonucleotides against
          a parenteral challenge of virulent B. anthracis spores.
          2) In guinea pigs, there appears to be some enhancement of specific protection against
          parenteral challenge. At 0 and 4 weeks guinea pigs were immunized with AVA or AVA + CpG (100 or
          300 micrograms). The animals were challenged i.m. at 10 weeks with about 5,000 Ames spores.
          Results:
          GROUP SURVIVED/TOTAL (% SURVIVAL)
          Saline Controls 1/28 (4)
          AVA 15/32 (47)
          AVA + CpG (100 micrograms) 12/15 (80)
          AVA + CpG (300 micrograms) 11/16 (69)
          ____________________________________________________________________
          3) Currently, at CBER, an experiment is being conducted to see whether CpG specifically
          enhances the serological immune response of rhesus monkeys to PA with both AVA and rPA vaccines.
          Animals have been immunized with AVA, AVA + CpG, rPA vaccine, or rPA vaccine + CpG. Periodic
          bleeds will be taken and serum tested by ELISA and TNA for anti-PA titers.
          – Bruce
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: Isoform studies information
          Date: Monday, September 10, 2001 2:28:33 PM
          Here is the information on the isoform studies. is writing the protocol. After
          determines the level of PA needed (2 doses) to give a mid-level protective response in rabbits, the
          same level of each of 2 PA isoforms will be used to immunize twice, at 0 and 4 weeks. At 10 weeks,
          the rabbits will be parenterally challenged with anthrax spores. Survival and anti-body titers will be
          determined. This experiment will hopefully demonstrate that the two isoforms are equally protective.
          This experiment is necessary in that two isoforms are found in purified preparations of rPA.
          – Bruce Ivins
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: RE: update for rPA
          Date: Tuesday, September 11, 2001 7:49:40 AM
          Thanks,
          – Bruce
          >—–Original Message—–
          >From:
          >Sent: Tuesday, September 11, 2001 7:29 AM
          >To:
          >Cc: Ivins, Bruce E Dr USAMRIID
          >Subject: update for rPA
          >
          >Here is the portion of the progress report from and myself including the rPA stability study and
          isoform investigation. I understand Bruce has already sent you the animal part of the isoform study.
          >
          >A contract is being finalized with Battelle to perform a combined stability and efficacy study. The GMP
          rPA product will be formulated with Alhydrogel at neutral pH (no formaldehyde) and held at three
          temperatures (4 C, 25 C, 40 C) for various time intervals (including 3 days; 1 week; 1,3,6 & 12
          months), after which rabbits will be immunized with a single dose of this material (0.5 ml containing
          100 ug protein) and then challenged with 100 LD50 of B. anthracis Ames spores. The route of exposure
          still under discussion will either be aerosol, to follow the ORD, or subcutaneous, for better control of the
          delivery and statistical evaluation of the results. Animals will be monitored for general signs of health
          and blood drawn at weekly intervals for bioassay of titer by ELISA and activity by TNA. Protein at each
          interval will also be evaluated for biophysical integrity by assaying rPA that is spontaneously and actively
          desorbed from the aluminum hydroxide.
          >
          >rPA isoforms will be purified by tandem anion exchange chromatography for use biological and
          biophysical tests. The composition of each of two isoforms will be investigated to identify the source of
          charge micro-heterogeneity. Amino acid modification including deamidation will be investigated using
          available methods including chemical stains and ESI-LC-MS/MS analysis of tryptic fragments.
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: FW: Anthrax, mice, and CpG
          Date: Tuesday, September 11, 2001 8:38:43 AM
          —–Original Message—–
          From:
          Sent: Tuesday, September 11, 2001 8:37 AM
          To: ‘Ivins Bruce E’
          Subject: RE: Anthrax, mice, and CpG
          Dear Bruce,
          I’ve been invited to deliver a lecture on the protective activity of CpG ODN
          in mid October. They’ve requested an abstract be submitted by the end of
          this week. Is the following OK with you?
          The ability of synthetic oligodeoxynucleotides (ODN) containing
          immunostimulatory “CpG” motifs to trigger an innate immune response capable
          of improving host survival following bacterial, viral and parasitic
          infection was investigated. Initial studies showed that normal mice treated
          with 5 – 50 ug of CpG ODN were protected for approximately 2 weeks from
          subsequent infection by >1,000 LD50 of Listeria monocytogenes or Francisella
          tularensis, or from 100 leishmania parasites. Subsequent studies
          established that higher doses of CpG ODN prolonged and/or enhanced the
          survival of mice challenged with biothreat agents such as inhaled anthrax
          and mouse-adapted Ebola virus. In each challenge model, protection was
          critically dependent on the CpG motif, being lost when the CpG dinucleotide
          was inverted or methylated.
          Two strategies were pursued to extend the protection conferred by
          CpG ODN. In the first, mice were treated with CpG ODN every two weeks for
          up to 4 months. Protection against L. monocytogenes and F. tularensis was
          induced and maintained for the duration of therapy. In the second, mice
          were injected with CpG ODN encapsulated in cationic stealth liposomes (CSL).
          CSL encapsulation significantly improved the uptake of ODN by cells of the
          immune system and increased the magnitude and persistence of the resultant
          immune response. Mice treated with encapsulated CpG ODN were protected
          from L. monocytogenes and F. tularensis for >4 weeks.
          To determine whether CpG ODN would be effective immunoprotective
          agents in humans, we examined the response of PBMC from >100 normal human
          donors to a large panel of ODN. Two distinct classes of CpG ODN were
          identified that activated human PBMC. The first (“K” type) ODN have a
          phosphorothioate backbone and multiple TCGTT and/or TCGTA. “K” ODN
          primarily stimulate B cells and monocytes to proliferate and secrete IgM,
          IL-6 and IL-10. The second (“D” type) ODN have a mixed phosphodiester/
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          phosphorothioate backbone and contain a single hexameric
          purine/pyrimidine/CG/purine/pyrimidine motif flanked by self-complementary
          bases that form a stem-loop structure capped at the 3′ end by a poly G tail.
          “D” ODN induce APC to mature and secrete IFNa and triggered NK cells to
          secrete IFNg.
          Whereas PBMC from all donors responded to at least one member of the
          “D” and “K” ODN panel, no single ODN induced an optimal response in all
          subjects. However, mixtures of ODN containing multiple CpG motifs strongly
          activated PBMC from all subjects. These mixtures also stimulated PBMC from
          non-human primates, such as chimpanzees and rhesus macaques. These mixtures
          were tested for their ability to protect rhesus monkeys from cutaneous
          leishmania infection. The severity of leishmania infection in macaques
          treated with “D” ODN 3 days before and 3 days after challenge was reduced by
          >90% (p. >
          >1. Vaccine: NEW recombinant PA (50 micrograms) + Dulbecco’s PBS + Alhydrogel (0.5 mg metallic aluminum) in a total volume of 0.5 ml, with or without formaldehyde (to be determined prior to experiment)
          >
          >2. Animals – New Zealand white rabbits, 2.5-3.5 kg and rhesus macaques, adults.
          >
          >3. Challenge – aerosol challenge with 100 – 500 LD50 of B. anthracis Ames spores.
          >
          ****************************************************************************************************************************************************************
          Immunization and challenge schedule for rabbits (10 males and 10 females per challenge group & 2 males and 2 females per control group):
          > 0 month and 1 month (4 weeks) – immunize 160 rabbits (8 groups of 20) with rPA vaccine, and immunize 8 control groups of rabbits with PBS
          > 3 months – challenge group 1 and group 1 controls
          > 6 months – challenge group 2 and group 2 controls
          > 12 months – challenge group 3 and group 3 controls
          > 12 months – immunize group 6 rabbits (with rPA vaccine) and group 6 controls (with PBS); if group 3 challenged animals have less than 80% (?) survival, challenge group 6 and group 6 control rabbits at 13 months
          > 18 months – challenge group 4 rabbits and group 4 controls
          > 18 months – immunize group 7 rabbits (with rPA vaccine) and group 7 controls (with PBS); if group 4 challenged animals have less than 80% (?) survival, challenge group 7 and group 7 control rabbits at 19 months
          > 24 months – challenge group 5 rabbits and group 5 controls
          > 24 months – immunize group 8 rabbits (with rPA vaccine) and group 8 controls (with PBS); if group 5 challenged animals have less than 80% (?) survival, challenge group 8 and group 8 control rabbits at 25 months
          >
          >5. Bleeding schedule for rabbits:
          > 1 week prior to 3 months – all rabbits
          > 4, 7, 10 and 14 days after challenge – group 1
          >>
          1 week prior to 6 months – all remaining rabbits
          > 4, 7, 10 and 14 days after challenge – group 2
          >>
          1 week prior to 12 months – all remaining rabbits
          > 4, 7, 10 and 14 days after challenge – group 3
          >>
          1 week prior to 13 months – group 6 and group 6 controls
          > 4, 7, 10 and 14 days after challenge – group 6
          >>
          1 week prior to 18 months – all remaining rabbits
          > 4, 7, 10 and 14 days after challenge – group 4
          >>
          1 week prior to 19 months – group 7 and group 7 controls
          > 4, 7, 10 and 14 days after challenge – group 7
          >>
          1 week prior to 24 months – all remaining rabbits
          > 4, 7, 10 and 14 days after challenge – group 5
          >>
          1 week prior to 25 months – group 8 and group 8 controls
          > 4, 7, 10 and 14 days after challenge – group 8
          >
          >6. Blood and sera will be assayed by ELISA, TNA, & ELISPOT.
          >*************************************************************************************************************************************************
          >7. Immunization and challenge schedule for monkeys (5 males and 5 females per challenge group & 1 male and 1 female per control group):
          >>
          0 month and 1 month (4 weeks) – immunize 77 monkeys (7 groups of 10) with rPA vaccine, and immunize 7 control groups with PBS
          > 6 months – challenge group 1 and group 1 controls
          > 12 months – challenge group 2 and group 2 controls
          > 12 months – immunize group 5 monkeys (with rPA vaccine) and group 5 controls (with PBS); if group 3 challenged animals have less than 80% (?) survival, challenge group 5 and group 5 control monkeys at 13
          months
          > 24 months – challenge group 3 rabbits and group 3 controls
          > 24 months – immunize group 6 monkeys (with rPA vaccine) and group 6 controls (with PBS); if group 5 challenged animals have less than 80% (?) survival, challenge group 6 and group 6 control monkeys at 25
          months
          > 36 months – challenge group 4 monkeys and group 4 controls
          > 36 months – immunize group 7 monkeys (with rPA vaccine) and group 7 controls (with PBS); if group 7 challenged animals have less than 80% (?) survival, challenge group 7 and group 7 control monkeys at 37
          months
          >>
          >8. Bleeding schedule for monkeys:
          >>
          1 week prior to 6 months – all monkeys
          > 4, 7, 10 and 14 days after challenge – group 1
          >>
          1 week prior to 12 months – all remaining rabbits
          > 4, 7, 10 and 14 days after challenge – group 2
          >>
          1 week prior to 13 months – group 5 and group 5 controls
          > 4, 7, 10 and 14 days after challenge – group 5
          >>
          1 week prior to 24 months – all remaining rabbits
          > 4, 7, 10 and 14 days after challenge – group 3
          >>
          1 week prior to 25 months – group 6 and group 6 controls
          > 4, 7, 10 and 14 days after challenge – group 6
          >>
          1 week prior to 36 months – all remaining rabbits
          > 4, 7, 10 and 14 days after challenge – group 4
          >>
          1 week prior to 37 months – group 7 and group 7 controls
          > 4, 7, 10 and 14 days after challenge – group 7
          >>
          >9. Blood and sera will be assayed by ELISA, TNA, & ELISPOT.
          >
          >***************************************************************************************************************************************************************************************
          >10. Surviving rabbits will be euthanized. Surviving monkeys will be “grayed out.”
          >
          >11. Rabbits will be challenged at 13, 19 or 25 months only if there is less than 80% survival at 12, 18 or 24 months.
          >
          >12. Monkeys will be challenged at 13, 25 or 37 months, only if there is less than 80% survival at 12, 24 or 36 months.
          >
          >***********************************************************************************************************************************************************************************
          >13, Points to consider:
          > a) Should part of the studies be contracted out in order to save USAMRIID animal room space? Where? SRI? Covance? SAIC?
          > b) Other points?
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          )
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          From: Ivins, Bruce E Dr USAMRIID
          To:
          Subject: RE: Anthrax, mice, and CpG
          Date: Tuesday, September 11, 2001 8:47:41 AM
          It looks fine. It’s very kind of you to include me on your abstract. I have only one comment:
          1) Mice were challenged with anthrax spores via the parenteral (subcutaneous) rather than the
          aerosol (inhaled) route.
          – Bruce
          —–Original Message—–
          From:
          Sent: Tuesday, September 11, 2001 8:37 AM
          To: ‘Ivins Bruce E’
          Subject: RE: Anthrax, mice, and CpG
          Dear Bruce,
          I’ve been invited to deliver a lecture on the protective activity of CpG ODN
          in mid October. They’ve requested an abstract be submitted by the end of
          this week. Is the following OK with you?
          The ability of synthetic oligodeoxynucleotides (ODN) containing
          immunostimulatory “CpG” motifs to trigger an innate immune response capable
          of improving host survival following bacterial, viral and parasitic
          infection was investigated. Initial studies showed that normal mice treated
          with 5 – 50 ug of CpG ODN were protected for approximately 2 weeks from
          subsequent infection by >1,000 LD50 of Listeria monocytogenes or Francisella
          tularensis, or from 100 leishmania parasites. Subsequent studies
          established that higher doses of CpG ODN prolonged and/or enhanced the
          survival of mice challenged with biothreat agents such as anthrax
          and mouse-adapted Ebola virus. In each challenge model, protection was
          critically dependent on the CpG motif, being lost when the CpG dinucleotide
          was inverted or methylated.
          Two strategies were pursued to extend the protection conferred by
          CpG ODN. In the first, mice were treated with CpG ODN every two weeks for
          up to 4 months. Protection against L. monocytogenes and F. tularensis was
          induced and maintained for the duration of therapy. In the second, mice
          were injected with CpG ODN encapsulated in cationic stealth liposomes (CSL).
          CSL encapsulation significantly improved the uptake of ODN by cells of the
          immune system and increased the magnitude and persistence of the resultant
          immune response. Mice treated with encapsulated CpG ODN were protected
          from L. monocytogenes and F. tularensis for >4 weeks.
          To determine whether CpG ODN would be effective immunoprotective
          (b) (6)
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          agents in humans, we examined the response of PBMC from >100 normal human
          donors to a large panel of ODN. Two distinct classes of CpG ODN were
          identified that activated human PBMC. The first (“K” type) ODN have a
          phosphorothioate backbone and multiple TCGTT and/or TCGTA. “K” ODN
          primarily stimulate B cells and monocytes to proliferate and secrete IgM,
          IL-6 and IL-10. The second (“D” type) ODN have a mixed phosphodiester/
          phosphorothioate backbone and contain a single hexameric
          purine/pyrimidine/CG/purine/pyrimidine motif flanked by self-complementary
          bases that form a stem-loop structure capped at the 3′ end by a poly G tail.
          “D” ODN induce APC to mature and secrete IFNa and triggered NK cells to
          secrete IFNg.
          Whereas PBMC from all donors responded to at least one member of the
          “D” and “K” ODN panel, no single ODN induced an optimal response in all
          subjects. However, mixtures of ODN containing multiple CpG motifs strongly
          activated PBMC from all subjects. These mixtures also stimulated PBMC from
          non-human primates, such as chimpanzees and rhesus macaques. These mixtures
          were tested for their ability to protect rhesus monkeys from cutaneous
          leishmania infection. The severity of leishmania infection in macaques
          treated with “D” ODN 3 days before and 3 days after challenge was reduced by
          >90% (p. —–Original Message—–
          > From:
          > Sent: Tuesday, September 11, 2001 8:46 AM
          > To:
          > Cc:
          > Subject: RE: rPA Rabbit serum
          >>
          I never heard anything from her after I sent her the email asking for any
          > suggestions (08/23/01). It was the rPA sera that was in the broken
          > bottles, correct?…did they supply this? We compared it side by side to
          > the normal sera, which was filterable. I was wondering if they have
          > noticed that it is so thick and how they work with it?
          >>
          We can try syringe filtering it with a 5.0 micrometer pore size (the
          > largest pore size we had been using was 0.4 micrometers) but I hate to
          > keep freezing and thawing it so I was hoping for some further direction
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          > from them before trying something else, so we could have multiple options
          > ready for the next time it is thawed. If they don’t have any ideas, we
          > can go ahead with syringe filtering it.
          >>
          >>
          —–Original Message—–
          > From:
          > Sent: Monday, September 10, 2001 5:14 PM
          > To:
          > Cc:
          > Subject: rPA Rabbit serum
          >>
          I talked to today. Did we resolve the issue with filtering
          > the broken sera? I told her we would also look at the rPA, normal, and
          > AVA sera to see if they all appear to be similar in consistency, color,
          > etc. the next time we have some thawed.
          >>
          >>
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        • DXer said

          I have posted Dr. Ivins’ emails from July, August and September 2001. Dr. Gerard Andrews, I think, explained that Dr. Ivins had a role in 19 different animal studies during this period.

          Civil depositions tend to involve a methodical Q and A about presented documents.

      • DXer said

        Perhaps one source of information relating to the series of experiments done in August, September and October would be REPORT OF PROTOCOL COMPETITION / TERMINATION available under FOIA from the VETERINARY MEDICINE DIVISION. FOIA officers need assistance in pinpointing to do ask and how to precisely describe the documents sought. Here is an example of a REPORT OF PROTOCOL COMPETITION / TERMINATION form relating to B99-97. (Dr. Ivins, his supervisor reported, was involved in 19 animal projects, I think).

        From:
        Sent: Wednesday, August 08, 2001 3:25 PM
        To: Ivins, Bruce E Dr USAMRIID
        Subject: Notification of Protocol Completion/Termination Form due for
        protocol:B99-07
        REPORT OF PROTOCOL COMPLETION / TERMINATION
        Date : 08/08/2001
        Division : BACTERIOLOGY
        Investigator : IVINS, BRUCE
        Protocol : B99-07
        TITLE : Protective Effect of Immunostimulatory Oligonucleotides Against Bacillus anthracis
        Intramuscular Challenge in Guinea
        1. Was the protocol been completed as
        orginally designed and/or ammemded?
        Explain: Yes
        2. Date completed: _25 JUN 01
        If terminated prior to completion, explain:
        3. Report any change to the number of animals
        in the painful procedure categories if different
        than the number reported in the approved protocol.
        No Pain = 48 (30%) Pain = 112 (70%) Originally we believed that 50% of the
        animals would be in the NO PAIN category
        4. What were the results and/or conclusions?
        (Report any negative findings if applicable.)
        The results indicated that specific immunity to anthrax spore challenge in guinea pig is enhanced
        by coadministration of CpG oligonucleotides.
        5. Will the research result in a publication or
        presentation? (Provide specifics.) We intend to present the data at the Annual Meeting of the
        American Society for Microbiology, Salt Lake City, Utah, in 2002.
        Date : 10 AUG 01 Signature/Division : __Bruce E. Ivins, USAMRIID Bacteriology Division_
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        RETURN THIS FORM TO (b) (6) VETERINARY MEDICINE DIVISION

        • DXer said

          I have submitted a FOIA to the wonderful USAMRC FOIA official:

          “Could I obtain a copy from Veterinary Medicine Division a copy of all REPORT OF PROTOCOL COMPETITION / TERMINATION for animal studies for which Dr. Bruce Ivins participated in FY 2001?

          One of the notebooks uploaded yesterday raises important issues regarding corroboration of how he spent his time in the B3. These Veterinary Medicine Division forms are an independent means of obtaining corroboration. It is the period of August 2001- December 2001 that is of interest but the forms might also be dated 2002, and refer back to earlier work.”

      • DXer said

        From: Ivins, Bruce E Dr USAMRIID To: Subject: “Old” formaldehyde experiments Date: Wednesday, September 12, 2001 9:47:56 AM Here is the information you requested on the Covance study: Five years ago we made rPA vaccine/Alhydrogel with and without formaldehyde added. We tested the vaccines after various periods of time of storage and noted (in guinea pigs) that the presence of formaldehyde appeared to boost potency of the vaccine. It was unknown whether the boost in potency was due to stabilization of the protein, or to an adjuvant effect. (Formaldehyde itself causes local inflammation which would draw APCs and other cell types to the site.) The vaccine is now 5 years old since it was formulated, and we wished to see (in the rabbit model) if there is any difference in potency between the 2 vaccines. (The rabbit model is preferred over the guinea pig model in tests of anthrax vaccine efficacy.) Twenty-four New Zealand white rabbits (12 of each gender) were immunized with 0.5-ml intramuscular doses of vaccine containing 50 micrograms rPA, Alhydrogel (0.5 mg aluminum) and PBS (with formaldehyde, 0.02%). Twenty-four New Zealand white rabbits (12 of each gender) were immunized with 0.5-ml intramuscular doses of vaccine containing 50 micrograms rPA, Alhydrogel (0.5 mg aluminum) and PBS (without formaldehyde). Four rabbits (2 of each gender) will be controls receiving Alhydrogel and PBS. Rabbits will be bled at weeks 2 and 4 for anti-PA antibody titers. They will be challenged subcutaneously with virulent anthrax spores 6 weeks after immunization and monitored for survival. This experiment will demonstrate whether the presence of formaldehyde in an rPA/Alhydrogel vaccine increases or preserves potency. – Bruce (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: CRM Task Order 0086 Date: Wednesday, September 12, 2001 1:26:29 PM I approve. – Bruce Ivins >—–Original Message—– >From: >Sent: Wednesday, September 12, 2001 1:26 PM >To: Ivins, Bruce E Dr USAMRIID >Cc: >Subject: CRM Task Order 0086 > >Dr. Ivins; > >I have received invoice no. 11 for for the month of August for $5,442.33. Please email reply to all your approval. Any questions, please call. > >Thanks 9/12/01 > (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Protocol modifications Date: Friday, September 14, 2001 10:32:09 AM – We need to wait until finishes his 2-dose study to decide what dose of rPA to use. If it turns out the dose is 10 micrograms -which is a good guess, will you have enough rPA for 2 shots, 30 rabbits of each isoform and 30 rabbits with rPA? Responses to 1. It is OK with me if we go to the larger number of rabbits. 2. OK. 3. This does not need to be a GLP study. This is still a tech-base study to tell us whether there are differences (as far as vaccinating ability) between the two isoforms. If it needs to be done GLP, then Battelle should do it. – Bruce >—–Original Message—– >From: >Sent: Tuesday, September 04, 2001 8:36 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: FW: Protocol modifications > >Re: animal protocol to study the contribution of rPA isoforms to protection against a lethal anthrax infection in rabbits. suggests we do 30 rabbits per group; see below. The change would result in 94 rabbits instead of 52 ($$$?). Please advise if we wish to accept his recommendations so I can make the changes. (Bruce had originally suggested 16 per group). Thanks in advance. > <> >> —–Original Message—– >From: >Sent: Friday, August 31, 2001 3:30 PM >To: >Subject: Protocol modifications >>> The isoform protocol is suggested to undergo the following statistical revisions. There are three reasons > for the revisions: >> 1. This study is approaching the final formulation of the rPA anthrax vaccine candidate and thus must > adhere to stricter criteria. >> 2. The stricter criteria involve the concept of “equivalency” testing. This means that we seek to reject > the hypothesis that the two isoforms are NOT equivalent with respect to survival and immune response > as measured by ELISA and TNA. Rejection of the hypothesis establishes the statistical evidence that the (b) (6) (b) (b) (6) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) > isoforms are equivalent. This requires larger sample sizes than a test of the hypothesis that the two > isoforms are not different (a subtle difference), which is an early developmental hypothesis. >> 3. Should this not be a strict GLP study due to the need for FDA approval of the decision that will > arise from it? We do not wish to have to repeat the study in the future under GLP standards. >> Revisions suggested: >> Page 3 replace objective with: “The hypothesis is that the two isoforms are NOT equivalent in survival rates > or immune response as measured by either ELISA and TNA. Equivalency is defined as a difference > in survival rates of no more than 20%, and a ratio of geometric mean ELISA titers and TNA titers of > no more than 4-fold respectively. The hypothesis will be tested at the 95% confidence level (2-tailed). Rejection > of the hypothesis establishes the equivalency of the survival and immune response of the two isoforms. > Failure to reject any of the hypothesis (survival or immune response) fails to establish the equivalency > of the two isoforms. Sex-specific tests will be done.” >> Page 11 replace Data Analysis with: “Sex-specific survival rates will be tested for equivalence using the method > of Farrington and Manning (ref). ELISA and TNA titers will be transformed to log10 and analyzed for > sex-specific equivalency based on the 95% confidence intervals (2-tailed) around the ratio of geometric mean titers. > Further analysis of survival correlations with sex, titers and isoform will use standard logistic regression. > Also, further analysis of titers will use multiple regression to test for sex differences and to test for > isoform differences. All tests will be at the 95% confidence level (2-tailed). All data will be automated > and verified prior to analysis. Statistical software package SAS (version 8.0 or greater) will be used > for analysis.” >> Page 11 Move the sample size justification to the page 5 and replace the statement there with: >> The test of equivalency of survival rates assumes 100% survival in both isoforms and a 20% > maximum difference resulting in a sample size of 15 of each sex per isoform. The test of > equivalency of immune response assumes a log10 standard deviation of 0.50 logs and a > maximum ratio of geometric mean titers of 4-fold (0.60 logs) resulting in a sample > size of 12 of each sex per isoform. Therefore, 30 rabbits (15 male and 15 female) must be > tested at each isoform and with the combined isoforms as a control. The unprotected controls > will be set at a nominal 4 animals (2 of each sex) due to confidence in the 100% lethality > of the challenge dose.” >> Reference: Farrington CP and Manning G. Test statistics and sample size formulae for comparative > binomial trials with null hypothesis of non-zero risk difference or non-unity relative risk. > Statistics in Medicine, vol 9, 147-1454, 1990. > > From: Ivins, Bruce E Dr USAMRIID To: ” Subject: RE: anti-rPA serum Date: Wednesday, September 19, 2001 8:13:16 AM Hi, , We will have both placebo and vaccine ready for pickup on Monday, September 24. We will make enough placebo to cover future immunizations as well, since there is no worry about antigen change or degradation. You may give my phone number or email out, but due to security precautions, they won’t be able to get on post. I will meet them at the Post office on 7th Street. They will come to Frederick via 270N, then continue on 15N. The exit they will take will be the 7th Street exit. They should bear to the right, and turn right onto 7th Street (towards Fort Detrick, not the hospital, which is towards the left.) They will go under the Route 15 overpass and take the first right into the post office parking lot. I will meet them there. If they need a map, I can send them one by FAX. – Sincerely, Bruce Ivins —–Original Message—– From: Sent: Tuesday, September 18, 2001 5:08 PM To: ‘Bruce.Ivins@DET.AMEDD.ARMY.MIL’ Subject: anti-rPA serum Bruce, It is time again to bother you. Sorry! Next booster of the 500 rPA-animals is scheduled for Tuesday, September 25 (6 micrograms in 0.5 ml doses). Our last meeting was so hectic (by the way, thanks much for all your help) that I forgot to request from you the placebo for the control group. This consists of 30 mice that will be bled in groups of ten each, three weeks after one, two or three doses of placebo (0.5 ml of same vaccine diluent containing same amount of aluminum than rPA-vaccine). Hopefully you can provide us with this latter prep this time (enough for all 30 mice; bleeding of the last group will be out of phase three weeks, but I don’t think it’s a big deal). One last thing. We are paying the contractor for picking up material; last time I delivered the vaccine myself in person, but in this instance I would prefer that Biocon picks it up directly from you. Is that possible? Please let me know, and if you don’t mind I will give them your phone to make arrangements for pick up directly. Thanks much and best regards. (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: UBMTA to sign Date: Wednesday, September 19, 2001 8:40:58 AM Thanks, – Bruce >—–Original Message—– >From: >Sent: Wednesday, September 19, 2001 8:40 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: UBMTA to sign > >Hi Bruce, >I received the signed UBMTAs from Whenever you get a chance can you please come by and sign & date the agreements. I’ll be at a meeting this morning. I’ll leave them on my chair. Before even receiving them, I started the routing initials (as you know it takes awhile). Once I get okay, we’re good to go. > (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: SPores Date: Thursday, September 20, 2001 9:41:06 AM If you think it would be helpful, I would be willing to come up to Battelle to offer advice or suggestions with respect to spore production, purification, storage, etc. When we first started working on it years ago, it took us quite awhile to learn the “art” and techniques involved in getting spores which were, stable, pure, unclumped, etc. Honestly, in 1 day to no more than a week, I might be able to save more than several weeks to several months worth of work on your part. Although spore production and purification sounds, on the surface, very easy and cookbook, there are some little nuances in the methodology which are important, yet hard to write down. Seriously, please let me know if you’d like me to come, and if so, when. At a time like this, I think we all need to come together in single directed purpose, and if that includes my coming there for one or a few days, I’d be happy to do it. – Bruce (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: Research Progress report Date: Thursday, September 20, 2001 9:52:09 AM Attachments: I have enclosed my research progress report, with changes made as required by the Anthrax Steering Committee. – Bruce (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: FW: SPores Date: Thursday, September 20, 2001 10:12:05 AM I just sent this to and I thought I’d send it to you. If you think it might be helpful for me to make a trip to Battelle for advice, suggestions, etc. on spore purification, harvest, storage, etc., I’d be happy to come. It’s as much an art as a science, and I could be there for whatever parts of the process you’d like. The purification on gradients is especially tricky. – Bruce >—–Original Message—– >From: Ivins, Bruce E Dr USAMRIID >Sent: Thursday, September 20, 2001 9:41 AM >To: >Subject: SPores > , > If you think it would be helpful, I would be willing to come up to Battelle to offer advice or suggestions with respect to spore production, purification, storage, etc. When we first started working on it years ago, it took us quite awhile to learn the “art” and techniques involved in getting spores which were, stable, pure, unclumped, etc. Honestly, in 1 day to no more than a week, I might be able to save more than several weeks to several months worth of work on your part. Although spore production and purification sounds, on the surface, very easy and cookbook, there are some little nuances in the methodology which are important, yet hard to write down. >> Seriously, please let me know if you’d like me to come, and if so, when. At a time like this, I think we all need to come together in single directed purpose, and if that includes my coming there for one or a few days, I’d be happy to do it. >> – Bruce (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Contract Mod Date: Thursday, September 20, 2001 10:20:26 AM Thanks, I also think that a parenteral challenge will be the best in this situation, having been involved in both parenteral and aerosol challenges. – Bruce —–Original Message—– From: Sent: Thursday, September 20, 2001 10:18 AM To: ‘ Cc: Ivins, Bruce E Dr USAMRIID; Subject: RE: Contract Mod We are completing the protocols and I plan to send them to you by tomorrow. They will not be in SOP form but instead in rougher form, as you call “study specific methods.” I want to preview for you the following changes not mentioned in the SOW: 1. Two methods for desorption of rPA will be used as follows. Sodium carbonate for protein to be assayed by SDS-PAGE, Western, and RP-HPLC. Sodium Phosphate for assay by native PAGE (Phast Gel). These procedures are identical except for the solution as indicated. The rationale is that while sodium carbonate will desorb much of the rPA its biophysical form is altered as seen by native PAGE but not by the other assays. Sodium phosphate will elute rPA that is identical to unadsorbed rPA, but the yield is only about 50%. So, we will look at the bulk of the adjuvanted protein with assays that are not sensitive to slight changes in charge, and look at a smaller portion of the adjuvanted protein using an assay that will distinguish between charge forms. 2. The tech base committee recommended injection rather than aerosol for exposure due to much greater precision in delivery of spores and subsequent confidence in statistical analysis of results. However, I am still awaiting a “go” from our prime systems contractor. I know this would change the costing and planning, so I will get back to you as soon as I get a final word from those in higher authority. —–Original Message—– From: Sent: Monday, September 17, 2001 1:20 PM To: ‘ Cc: Subject: Contract Mod Hi On Friday we received the contract mod from the USAMRMC Contracting Officer to start your stability project. We have started to write the animal use protocol and we ordered the PhastSystem today. Any idea when you can provide us with all the SOPs and Methods that we need to have in place (in our format) before we can start the work? We also need to schedule the first teleconference – perhaps a better use of everybody’s time if we schedule it after you have sent us the SOPs and we have had a couple of days to review them so that we can ask relevant questions. Thanks (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Cc: Subject: rPA Date: Thursday, September 20, 2001 2:41:41 PM , Thank you for the 30 vials of rPA, lot 100506, 1.18 mg/ml, that you gave me this morning for tech-base studies in support of the development of a new human anthrax vaccine. It will be put to use immediately. – Bruce Ivins (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Cc: Subject: B98-03 rabbit bacteremia data Date: Tuesday, September 25, 2001 9:13:34 AM Attachments: Here are the B98-03 rabbit bacteremia data in an EXCEL file. – Bruce (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: Money for long-term rabbit study Date: Tuesday, September 25, 2001 11:22:04 AM This is the money I come up with for the long-term rabbit study. What is needed for the long-term monkey study and the strain study will be forthcoming. The figures are for supplies, animals, etc. necessary to do the study. Personnel money (salaries, travel, etc.) is not included. Costs: 192 rabbits…………………………………………………$21,120 Aerosolization costs………………………………………$1,920 Supplies costs (Tyveks, gloves, booties, head covers, syringes, needles, tubes…………………$10,000 ELISA and TNA costs…………………………………….$3,400 Husbandry costs (daily rabbit care)…………………$296,640 __________ Total………………………………………………………..$330,080 ********************************************************************************************************************************************************************** as just given me her costs for the diverse strain study: Costs: 200 rabbits………………………………………………..$22,000 Animal husbandry (100 days)…………………………$60,000 Aerosolization costs………………………………………$2,000 Supplies……………………………………………………$10,000 ELISA and TNA assays………………………………….$1,000 _______ Total………………………………………………………..$95,000 I’ll get the monkey study info to you as soon as possible. – Bruce (b) (6) (b) (6) (b ) (6 ) From: Ivins, Bruce E Dr USAMRIID To: Subject: Money for long-term rabbit study Date: Tuesday, September 25, 2001 11:22:04 AM This is the money I come up with for the long-term rabbit study. What is needed for the long-term monkey study and the strain study will be forthcoming. The figures are for supplies, animals, etc. necessary to do the study. Personnel money (salaries, travel, etc.) is not included. Costs: 192 rabbits…………………………………………………$21,120 Aerosolization costs………………………………………$1,920 Supplies costs (Tyveks, gloves, booties, head covers, syringes, needles, tubes…………………$10,000 ELISA and TNA costs…………………………………….$3,400 Husbandry costs (daily rabbit care)…………………$296,640 __________ Total………………………………………………………..$330,080 ********************************************************************************************************************************************************************** as just given me her costs for the diverse strain study: Costs: 200 rabbits………………………………………………..$22,000 Animal husbandry (100 days)…………………………$60,000 Aerosolization costs………………………………………$2,000 Supplies……………………………………………………$10,000 ELISA and TNA assays………………………………….$1,000 _______ Total………………………………………………………..$95,000 I’ll get the monkey study info to you as soon as possible. – Bruce (b) (6) (b) (6) (b ) (6 ) From: Ivins, Bruce E Dr USAMRIID To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Money for long-term rabbit study Date: Tuesday, September 25, 2001 4:07:00 PM Monkey long-term efficacy study money requirements. Please keep in mind that the costs are for 3 years. We plan to use 10 monkeys + 2 controls per time point (3 months, 6 months, 12 months, 24 months, 25 months (if 24 month survival is less than 80%), 36 months, and 37 months (if 36 month survival is less than 80%). Cost: 72 monkeys….($3,000 per monkey)………………………….$216,000 Aerosolization costs…………………………………………………$1,000 Supplies………………………………………………………………$10,000 ELISA and TNA costs……………………………………………….$2,000 Husband costs ($6.00 per day per animal)…………………..$822,240 __________ Total………………………………………………………………..$1,051,240 If the costs are deemed too high, then we can lower the number of timepoints, thus lowering the total number of monkeys. – Bruce >—–Original Message—– >From: Ivins, Bruce E Dr USAMRIID >Sent: Tuesday, September 25, 2001 11:22 AM >To: Ivins, Bruce E Dr USAMRIID; >Subject: Money for long-term rabbit study > This is the money I come up with for the long-term rabbit study. What is needed for the long-term monkey study and the strain study will be forthcoming. The figures are for supplies, animals, etc. necessary to do the study. Personnel money (salaries, travel, etc.) is not included. > >Costs: > >192 rabbits…………………………………………………$21,120 >Aerosolization costs………………………………………$1,920 >Supplies costs (Tyveks, gloves, booties, head > covers, syringes, needles, tubes…………………$10,000 >ELISA and TNA costs…………………………………….$3,400 >Husbandry costs (daily rabbit care)…………………$296,640 > __________ >Total………………………………………………………..$330,080 >> >********************************************************************************************************************************************************************** has just given me her costs for the diverse strain study: > >Costs: > >200 rabbits………………………………………………..$22,000 >Animal husbandry (100 days)…………………………$60,000 >Aerosolization costs………………………………………$2,000 >Supplies……………………………………………………$10,000 >ELISA and TNA assays………………………………….$1,000 > _______ >Total………………………………………………………..$95,000 >>> >I’ll get the monkey study info to you as soon as possible. >- Bruce (b) (6) (b) (6) (b) (6) ( b ) ( 6 ) (b ) (6) From: Ivins, Bruce E Dr USAMRIID To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Money for long-term rabbit study Date: Tuesday, September 25, 2001 4:07:00 PM Monkey long-term efficacy study money requirements. Please keep in mind that the costs are for 3 years. We plan to use 10 monkeys + 2 controls per time point (3 months, 6 months, 12 months, 24 months, 25 months (if 24 month survival is less than 80%), 36 months, and 37 months (if 36 month survival is less than 80%). Cost: 72 monkeys….($3,000 per monkey)………………………….$216,000 Aerosolization costs…………………………………………………$1,000 Supplies………………………………………………………………$10,000 ELISA and TNA costs……………………………………………….$2,000 Husband costs ($6.00 per day per animal)…………………..$822,240 __________ Total………………………………………………………………..$1,051,240 If the costs are deemed too high, then we can lower the number of timepoints, thus lowering the total number of monkeys. – Bruce >—–Original Message—– >From: Ivins, Bruce E Dr USAMRIID >Sent: Tuesday, September 25, 2001 11:22 AM >To: Ivins, Bruce E Dr USAMRIID; >Subject: Money for long-term rabbit study > > This is the money I come up with for the long-term rabbit study. What is needed for the long-term monkey study and the strain study will be forthcoming. The figures are for supplies, animals, etc. necessary to do the study. Personnel money (salaries, travel, etc.) is not included. > >Costs: > >192 rabbits…………………………………………………$21,120 >Aerosolization costs………………………………………$1,920 >Supplies costs (Tyveks, gloves, booties, head > covers, syringes, needles, tubes…………………$10,000 >ELISA and TNA costs…………………………………….$3,400 >Husbandry costs (daily rabbit care)…………………$296,640 > __________ >Total………………………………………………………..$330,080 >> >********************************************************************************************************************************************************************** has just given me her costs for the diverse strain study: > >Costs: > >200 rabbits………………………………………………..$22,000 >Animal husbandry (100 days)…………………………$60,000 >Aerosolization costs………………………………………$2,000 >Supplies……………………………………………………$10,000 >ELISA and TNA assays………………………………….$1,000 > _______ >Total………………………………………………………..$95,000 >>> >I’ll get the monkey study info to you as soon as possible. >- Bruce (b) (6) (b) (6) (b) (6) (b) (6) (b ) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: Strains Date: Monday, October 01, 2001 2:41:38 PM Hi, Thanks for your phone call earlier today. and I both gave some thought as to the B. anthracis strains of interest with respect to sequencing. Am I correct that the Ames strain has been sequenced? It certainly would be a fine candidate for a “type strain.” The other strains would be 1) Vollum 1B 2) The Australia strain that you and sent (It killed all 16 immunized guinea pigs.) 3) The Kruger A isolate 4) The Kruger B isolate I think that if you do one of the Kruger isolates, you should do the other. The Vollum 1B strain is of interest because it has been so extensively used, and because it is demonstrably less virulent than the Ames strain in certain animal models. The Australia strain is of interest because of its exceptional virulence in the immunized guinea pig. Hope this is helpful. If you’d like to talk about these any more, I’d be happy to do so. Have a great fall! – Bruce (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: Florida case(?) Date: Thursday, October 04, 2001 9:57:19 PM Hi, I just heard this evening (and read over internet news) that a case of pulmonary anthrax may have been identified in Florida. Is this true, or is this just hysteria? The only Florida strain of B. anthracis that I am familiar with is V770, which is the parent of V770-NP1-R, the strain used in production of the human anthrax vaccine. (I believe that V770 was originally isolated from a cow in Florida in the early 1950s.) The article said that this person was an “Outdoorsman,” and had drunk water from a creek in North Carolina. If he really does have anthrax, could he have gotten it this way, or did he get it by tromping around some dusty field area. (Has North Carolina been dry this summer?) I know that in the wild in Africa, animals are supposed to be able to get it from water holes by stirring up spores and presumably ingesting and possibly inhaling them as an aerosol. Could this have happened? What if an animal had died upstream and the stream was contaminated? (Drinking from a stream or creek without boiling or purifying the water first is an invitation to intestinal disease or parasites, but have any other human anthrax cases been documented from people drinking contaminated water?) You called me several times in the recent past with regards to another anthrax issue. If there’s anything I can help with here (if you or coworkers are involved) please let me know. I don’t know if there’s anything I can do, but I’m certainly willing to provide whatever informational assistance I can. (I would have been less surprised if the Florida man had been hunting deer in Texas, where there is identifiable anthrax. I don’t recall North Carolina as having ideal soil for preservation of anthrax spores or for anthrax cycling of spore-vegetative cell-spore-vegetative cell etc., but I suppose there could be areas of higher soil calcium and alkalinity.) Anyway, please don’t hesitate to give me a call if there’s anything I can do. We are currently testing the virulence (in immunized and unimmunized guinea pigs) of B. anthracis strains from all over the world, including China, and we’ve come up with some very interesting differences in virulence among the strains. Take care of yourself, – Bruce (b) (6) (b) (6) (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 10:52:45 AM The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. Basically what we have as far as the experiment: 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. 2) Results so far, 3 days after challenge: Group Survived/Total A – Vaccine plus formaldehyde 24/24 (no deaths) B – Vaccine minus formaldehyde 16/24 (8 deaths) C – Controls 0/4 (4 deaths) Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. – Bruce (b) (6) (b) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:04:56 AM No, just survival data. – Bruce —–Original Message—– From: Sent: Friday, October 05, 2001 10:56 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine Bruce, Do you have any data yet on the protein composition (i.e. is it degraded) in the with or without formaldehyde? —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 10:53 AM To: Subject: Stabilizer in a new rPA vaccine The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. weren’t some FDA-acceptable stabilizers going to be identified? If (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) there some out there, maybe we should start thinking about them now. Basically what we have as far as the experiment: 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. 2) Results so far, 3 days after challenge: Group Survived/Total A – Vaccine plus formaldehyde 24/24 (no deaths) B – Vaccine minus formaldehyde 16/24 (8 deaths) C – Controls 0/4 (4 deaths) Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. – Bruce From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:04:56 AM No, just survival data. – Bruce —–Original Message—– From: Sent: Friday, October 05, 2001 10:56 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine Bruce, Do you have any data yet on the protein composition (i.e. is it degraded) in the with or without formaldehyde? —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 10:53 AM To: Subject: Stabilizer in a new rPA vaccine The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. and .weren’t some FDA-acceptable stabilizers going to be identified? If (b) (6) (b) (6) (b) (6) (b ) (6 ) (b) (6) (b) (6) (b) (b) (6) (6) there some out there, maybe we should start thinking about them now. Basically what we have as far as the experiment: 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. 2) Results so far, 3 days after challenge: Group Survived/Total A – Vaccine plus formaldehyde 24/24 (no deaths) B – Vaccine minus formaldehyde 16/24 (8 deaths) C – Controls 0/4 (4 deaths) Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. – Bruce From: Ivins, Bruce E Dr USAMRIID To: Ivins, Bruce E Dr USAMRIID; Subject: FW: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:07:04 AM It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. – Bruce >—–Original Message—– >From: >Sent: Friday, October 05, 2001 11:05 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: Stabilizer in a new rPA vaccine > >Bruce, >This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? >> —–Original Message—– >From: Ivins, Bruce E Dr USAMRIID >Sent: Friday, October 05, 2001 10:53 AM >To: >Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. >> – Bruce > From: Ivins, Bruce E Dr USAMRIID To: Ivins, Bruce E Dr USAMRIID; Subject: FW: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:07:04 AM It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. – Bruce >—–Original Message—– >From: >Sent: Friday, October 05, 2001 11:05 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: Stabilizer in a new rPA vaccine > >Bruce, >This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? >> —–Original Message—– >From: Ivins, Bruce E Dr USAMRIID >Sent: Friday, October 05, 2001 10:53 AM >To: >Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. >> – Bruce > From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:10:17 AM I don’t have an earlier time point in rabbits, just guinea pigs. – Bruce >—–Original Message—– >From: >Sent: Friday, October 05, 2001 11:10 AM >To: Ivins, Bruce E Dr USAMRIID >Subject: RE: Stabilizer in a new rPA vaccine > >Do you have an earlier time point? Is this the same batch of protein that showed to be more degraded with formaldehyde than without? >> —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, October 05, 2001 11:07 AM > To: Ivins, Bruce E Dr USAMRIID; > Subject: FW: Stabilizer in a new rPA vaccine >> It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. > – Bruce > —–Original Message—– > From: > Sent: Friday, October 05, 2001 11:05 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: RE: Stabilizer in a new rPA vaccine >> Bruce, > This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? > >> —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, October 05, 2001 10:53 AM > To: > Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. >> – Bruce > From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:21:42 AM That is domain. – Bruce —–Original Message—– From: Sent: Friday, October 05, 2001 11:02 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine You will still need the protein data to confirm that it is a stability problem and not an adjuventing affect. (yeah, I know, I always have to play devil’s advocate). —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 11:07 AM To: Ivins, Bruce E Dr USAMRIID; Subject: FW: Stabilizer in a new rPA vaccine It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. – Bruce > —–Original Message—– > From: (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) > Sent: Friday, October 05, 2001 11:05 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: RE: Stabilizer in a new rPA vaccine >> Bruce, > This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? > >> —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, October 05, 2001 10:53 AM > To: > Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. and .weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. > (b) (6) (b) (6) (b) (b) (6) (6) > – Bruce > From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:21:42 AM That is domain. – Bruce —–Original Message—– Sent: Friday, October 05, 2001 11:02 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine You will still need the protein data to confirm that it is a stability problem and not an adjuventing affect. (yeah, I know, I always have to play devil’s advocate). —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 11:07 AM To: Ivins, Bruce E Dr USAMRIID; Subject: FW: Stabilizer in a new rPA vaccine It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. – Bruce > —–Original Message—– > From: (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) > Sent: Friday, October 05, 2001 11:05 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: RE: Stabilizer in a new rPA vaccine >> Bruce, > This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? >> —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, October 05, 2001 10:53 AM > To: > Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. and ..weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. > (b) (6) (b) (6) (b) (b) (6) (6) > – Bruce > From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:31:44 AM The only data we have on this is the guinea pig data. Perhaps the question can be addressed by the stability study at Battelle. .your thoughts??? – Bruce —–Original Message—– From: Sent: Friday, October 05, 2001 11:21 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine If all or most of the rabbits die in the without formaldehyde group, then this would more strongly suggest that you need a stabilizer. Still, the vaccine is 5 years old; and if the rPA did degrade, we don’t know anything about the rate of degradation or how this correlates to efficacy. For the licensed vaccine, the ORD says it needs to be stable for 2 years. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 11:22 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine That is domain. – Bruce (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) —–Original Message—– From: Sent: Friday, October 05, 2001 11:02 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine You will still need the protein data to confirm that it is a stability problem and not an adjuventing affect. (yeah, I know, I always have to play devil’s advocate). —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 11:07 AM To: Ivins, Bruce E Dr USAMRIID; Subject: FW: Stabilizer in a new rPA vaccine It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. – Bruce > —–Original Message—– > From: > Sent: Friday, October 05, 2001 11:05 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: RE: Stabilizer in a new rPA vaccine >> Bruce, > This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? >>> —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, October 05, 2001 10:53 AM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) > To: > Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. and weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. >> – Bruce > (b) (6) (b) (b) (6) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 11:31:44 AM The only data we have on this is the guinea pig data. Perhaps the question can be addressed by the stability study at Battelle. …your thoughts??? – Bruce —–Original Message—– From: Sent: Friday, October 05, 2001 11:21 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine If all or most of the rabbits die in the without formaldehyde group, then this would more strongly suggest that you need a stabilizer. Still, the vaccine is 5 years old; and if the rPA did degrade, we don’t know anything about the rate of degradation or how this correlates to efficacy. For the licensed vaccine, the ORD says it needs to be stable for 2 years. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 11:22 AM To: Subject: RE: Stabilizer in a new rPA vaccine That is domain. – Bruce (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) —–Original Message—– From: Sent: Friday, October 05, 2001 11:02 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine You will still need the protein data to confirm that it is a stability problem and not an adjuventing affect. (yeah, I know, I always have to play devil’s advocate). —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 11:07 AM To: Ivins, Bruce E Dr USAMRIID; Subject: FW: Stabilizer in a new rPA vaccine It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. – Bruce > —–Original Message—– > From: > Sent: Friday, October 05, 2001 11:05 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: RE: Stabilizer in a new rPA vaccine >> Bruce, > This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? >>> —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, October 05, 2001 10:53 AM (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) > To: > Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. and .weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. >> – Bruce > (b) (6) (b) (b) (6) (6) From: Ivins, Bruce E Dr USAMRIID To: Subject: RE: Stabilizer in a new rPA vaccine Date: Friday, October 05, 2001 2:29:40 PM All – After the final data are in, shall we plan a meeting to discuss where we (tech base/advanced development) go from here with respect to incorporating a stabilizer in the vaccine? Possible questions to address: 1) What if any studies are necessary to follow up these data? 2) What stabilizers are acceptable to the FDA? 3) What path should we take as far as incorporating a particular stabilizer into a new anthrax vaccine? 4) Others(???) – Bruce —–Original Message—– From Sent: Friday, October 05, 2001 12:17 PM To: Subject: RE: Stabilizer in a new rPA vaccine In my opinion, this shows that half of the rPA had spontaneously desorbed from the Alhydrogel after 5 years, and remained undegraded in the presence of formaldehyde (lane 9), or completely degraded in the absence of formaldehyde (lane 8). Protein that remained on the Alhydrogel multimerized in the presence of formaldehyde (lane 4) or remained largely intact in the absence of formaldehyde, while at the same time having a larger number of breakdown products of small concentration (lane 5). Both of the latter samples contain the familiar charge isoforms of full-length rPA protein. —–Original Message—– From: Sent: Friday, October 05, 2001 11:51 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine Here is the gel on rPA desorbed from 5 year old vaccine. —–Original Message—– From: Ivins, Bruce E Dr USAMRIID Sent: Friday, October 05, 2001 11:22 AM To: (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) Subject: RE: Stabilizer in a new rPA vaccine That is domain. – Bruce —–Original Message—– From: Sent: Friday, October 05, 2001 11:02 AM To: Ivins, Bruce E Dr USAMRIID; Subject: RE: Stabilizer in a new rPA vaccine You will still need the protein data to confirm that it is a stability problem and not an adjuventing affect. (yeah, I know, I always have to play devil’s advocate). —–Original Message—– From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL] Sent: Friday, October 05, 2001 11:07 AM To: Ivins, Bruce E Dr USAMRIID; Subject: FW: Stabilizer in a new rPA vaccine It’s up to everyone else. It does strongly appear as though we will need a stabilizer, however. – Bruce > —–Original Message—– > From: > Sent: Friday, October 05, 2001 11:05 AM > To: Ivins, Bruce E Dr USAMRIID > Subject: RE: Stabilizer in a new rPA vaccine (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) (b) (6) >> Bruce, > This could figure into the upcoming stability/efficacy study, for which I specifically asked about an concurrent set with an excipient but DVC considered this too early. Should we meet? >> —–Original Message—– > From: Ivins, Bruce E Dr USAMRIID > Sent: Friday, October 05, 2001 10:53 AM > To: > Subject: Stabilizer in a new rPA vaccine >> The data we are getting from our with formaldehyde/without formaldehyde experiment in rabbits is giving us VERY strong evidence that we should incorporate a stabilizer in with rPA and Alhydrogel. ..weren’t some FDA-acceptable stabilizers going to be identified? If there some out there, maybe we should start thinking about them now. >> Basically what we have as far as the experiment: >> 1) Five years ago rPA/Alhydrogel/PBS vaccine was made with or without 0.02% formaldehyde (the level that’s in AVA) and stored at 4C. With these vaccines we immunized groups of rabbits as follows (0.5 ml per intramuscular dose): > Group A – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/0.02% formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group B – 24 rabbits (12 males, 12 females) get PA (50 ug)/Alhydrogel (0.5 mg)/PBS/No formaldehyde at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. > Group C – 4 rabbits (2 males, 2 females) get PBS/Alhydrogel (0.5 mg) at 0 weeks. Challenge (subcutaneous) at 6 weeks with about 100 LD50 Ames spores. >> 2) Results so far, 3 days after challenge: >>> Group Survived/Total >> A – Vaccine plus formaldehyde 24/24 (no deaths) >> B – Vaccine minus formaldehyde 16/24 (8 deaths) >> C – Controls 0/4 (4 deaths) >>> Note: We originally studied the effect of formaldehyde on rPA vaccine potency/stability in guinea pigs. The cumulative data indicated that stability/potency was enhanced by the presence of formaldehyde. >> – Bruce > (b) (6) (b) (6) (b) (6)
  2. Lew Weinstein said

    I seem to recall the FBI/DOJ saying it had presented material to a grand jury and was about to seek an indictment when Ivins committed suicide. Can this be confirmed? Or is it part of a “created story” invented to try to close the case on Aug 8, 2008?

  3. anonymous said

    http://www.thedailybeast.com/blogs-and-stories/2011-06-02/the-mirage-man-by-david-willman-anthrax-attacker-bruce-ivinss-obsessions/#
    dwurry
    In reality, Bruce Ivins was never accused of a crime while he was alive. The FBI concluded after uncovering loads of dirt on him that since he killed himself he must be guilty. In reality, the stuff the FBI dug up on him destroyed his family, his relationship with his children and probably lead to his suicide. We value privacy and freedom in this country–or we used to–because that is your best defense against improper investigation.

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