Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
Contract No. HHSN266200500040-C
ADB Contract No. N01-AI-50040
Section I: Purpose and Scope of Effort
The Tularemia Vaccine Development Contract will lead to vaccine candidates, two animal
models and cellular assays vital for testing vaccine efficacy.
Sections II and III: Progress and Planning Presented by Milestone
Active milestones: 1, 2, 3, 5, 12, 34, 41, 46
Inactive milestones: 4, 6-11, 13-33, 35-40, 42-45, 47-54
Milestone 1
Milestone description: Subcontracts finalized
Institution: UNM/All subcontractors
1. Date Started: 11/1/2005
2. Date Completed: pending
3. Work performed and progress including data and preliminary conclusions
a. LBERI, ASU, Cerus and UTSA have signed contracts with COA numbers assigned. LBERI
b.
c.
d.
e.
f.
g.
h.
has COA#2 as of 2/23/06, ASU has COA#3 as of 3/2/06, Cerus has COA#4 as of 3/2/06
and UTSA has COA#6 as of 4/3/06.
Final fully executed subcontracts for Cerus, ASU, and LBERI have been sent to Ross
Kelley within 30 days of the COA assignment date. UNM is awaiting UTSA’s signed
contract to arrive so that UNM can sign and send to Ross Kelley.
Cerus, ASU, LBERI and UTSA have received formatted templates for invoices to UNM, for
monthly Financial Reports by Milestone, for monthly Technical Reports, and for monthly
detail documentation back up for Materials and Supplies Expenses (M&S).
Rick Lyons and Barbara Griffith traveled to Cerus for a site on 3/22/2006 for one day to
setup financial, technical and timeline reporting expectations and to assure quality for the
scientific milestones.
UNM executed NDAs with all 3 DVC subcontractors (Umea University, NRC, and DSTL)
before the 3/28/06 meeting in Charleston, SC.
UNM participated in a joint conference with NIAID and DVC in Charleston, SC 3/28 to
3/30/06 to open communication between the two TVDC teams.
UNM Strategic Work Plan: re-revised version was provided to Contract and Project Officers
on 4/5/06 and review is anticipated by 4/12/06.
Barbara, Mindy and Lori Diehl have conferred with two MS Project Consultants who
recommended that UNM not divide the MS Project file into subunits by contractor as the
reassembly into a unified MS Project file would be problematic for linkages and dates.
4. Significant decisions made or pending
a. The customized timeline report template in MS Project is being reconsidered. UNM is now
planning to retain the MS Project timeline file and have subcontractors submit timeline
changes to UNM so that UNM can manually enter the changes into MS Project to retain the
linkages and dates.
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
5. Problems or concerns and strategies to address
Mindy is taking a 6 session MS Project course and Barbara and Rick are contemplating a small
contract with an MS Project Consultant (Ms Jane Betterton) utilizing discretionary, non-TVDC
funds. The goal is to develop efficient reports from the MS Project timeline file and to develop a
template for timeline data transmission from the subcontractors to UNM. Data will be manually
entered at UNM into the MS Project file to retain linkages and date consistency.
6. Deliverables completed
3 fully executed subcontracts have been finalized and returned to Ross Kelley (Cerus, ASU, and
LBERI)
7. Quality of performance
Excellent
8. Percentage completed
90%
9. Work plan for upcoming month
a.
b.
c.
d.
UNM to complete Strategic Work Plan after 4/12/06 review by Vicki Pierson
UNM to send Ross Kelley fully signed and executed subcontract with UTSA by ~ 5/3/06
UNM to finalize strategy for timeline changes to be reported by subcontractors to UNM
UNM to add the changes to the MS Project timeline file.
10. Anticipated travel
Dr. Lyons and Barbara Griffith are planning site visits to ASU and UTSA at the end of April to
setup financial, technical and timeline reporting expectations and to assure quality for the
scientific milestones.
11. Upcoming Contract Authorization (COA) for subcontractors
a. COA#5 4/03/06 approved purchase of equipment for monitoring blood coagulation
parameters in the ABSL facility, the complete bronchscopy system and the biobubble for
LBERI usage.
b. COA#6- 4/3/06 approved subcontract with UTSA
c. Barbara will submit requests for COAs for approved equipment on the Cerus subcontract
and approved equipment on the ASU subcontract; Cerus obtained current quotations from
equipment vendors prior to seeking approval to purchase.
d. Barbara will submit a request for COA for new UNM conference phone equipment and web
site software site licenses.
Milestone 2
Milestone description: Vaccinations performed on relevant personnel
Institution: UNM/LRRI
1. Date started: 11/01/1005
2. Date completed: pending
3. Work performed and progress including data and preliminary conclusions
NIAID is working on the IAA with USAMRIID and discussions regarding funding transfers are in
progress.
4. Significant decisions made or pending
a. As of 3/6/2006 NIAID is waiting for Martin Crumrine, DMID Program Officer, to provide
an update on the status of the IAA between NIAID and USAMRIID.
b. UNM has notified NIAID that if vaccinations are not administered by February 2006, or no
later than June 2006, additional protective equipment will be necessary to prevent a
stoppage of work.
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
c. The number of potential vaccinees has increased to 41 for LBERI, 6 for UTSA, 4 for
UNM. Total = 51, though LBERI originally had 20 our on TVDC and had ~21 on a project
associated with Judy Hewitt. UNM currently anticipates funding the travel for 32
scientists (22 from LBERI, 6 for UTSA and 4 for UNM) to USAMRIID and anticipates
funding the pre-health screenings for 26 (22 from LBERI and 4 for UNM) since UTSA is
paying for their own pre-health screenings. UNM EOHS has provided a quote for
$579/person for the pre-health screenings including chest X-ray, infectious disease
testing and EKG.
d. The “Occupational Health budget” is approximately $341,711. The costs for UNM
prehealth screenings are now estimated at cost $579/person and include labs, chest xray, EKG (see list at end of agenda); same cost for UNM and LBERI employees. 26 x
$579=$15,054
i. Travel & labor has been estimated at min of $3400/person.
ii. So prehealth/travel/labor= min of ~$4000/person. 32 x $4000= $128,000
iii. If travel /labor /prehealth is closer to $8,000/person 32 x $8000= $256,000
e. UNM received COA#5 to purchase the biobubble for LBERI as additional protective
equipment.
5. Problems or concerns and strategies to address
When will the NIAID-USAMRIID IAA be completed and when can UNM begin the prehealth
screenings?
6. Deliverables completed
None
7. None Quality of performance
Good
8. Percentage completed
12%
9. Work plan for upcoming month
Ross Kelley will continue to monitor the progress of whether Martin Crumrine's IAA between
NIAID and USAMRIID will inform UNM when and whether the TVD Contractors can be
vaccinated under this IAA.
10. Anticipated travel
None in the next month; travel could occur in any month between May and June 2006
11. Upcoming Contract Authorization (COA) for subcontractors
None
Milestone 3
Milestone description: Bioaerosol technique selected and optimized
Institution: LBERI
1. Date started: 2/23/2006
2. Date completed: in progress
3. Work performed and progress including data and preliminary conclusions
Submitted ES&H work permit for approval
4. Significant decisions made or pending
a. Will use Collison nebulizer as the baseline for comparison
b. Selected 2 additional bioaerosol generators for comparison testing; liquid sparging
generator and micropump generator
5. Problems or concerns and strategies to address
None
6. Deliverables completed
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
None
7. Quality of performance
Fair
8. Percentage completed
1%
9. Work plan for upcoming month
a. Order and obtain first 2 pieces of equipment; micropump and liquid sparging generators
b. Obtain ES&H authorization
c. Obtain LVS vials from UNM
10. Anticipated travel
None anticipated at the present time
11. Upcoming Contract Authorization (COA) for subcontractors
Preparing COA for liquid sparging generator
Milestone 5
Milestone description: Species tested for sensitivity to LVS & generation of immunity against a
pulmonary challenge of Schu4
Institution: UNM
1. Date started: 12/12/2005
2. Date completed: pending
3. Work performed and progress including data and preliminary conclusions
a. We challenged the mice that had survived LVS inoculation in experiment Ftc6
(henceforth called vaccinated mice) intranasally with SCHU S4 to determine the
vaccination route and dose that would generate the best protection against i.n. SCHU S4
challenge. The experiment code is Ftc6B
i. Based on our experience with vaccinated BALB/c mice, we wanted to challenge
the vaccinated mice intranasally with an sublethal dose of 2 x 102 CFU and a
lethal dose of 2 x 103 CFU SCHU S4. However, the concentration of SCHU S4
in the inocula was 60% lower than targeted and consequently the number of
bacteria actually deposited in the deep lung was 28 CFU and 293 CFU.
ii. Intranasal vaccination generated the best protection in NIH-Swiss mice (Fig. 1).
Eleven days after i.n. challenge with 293 CFU SCHU S4, 25% of the i.n.
vaccinated mice had died but more than 60% of the s.c.- and i.d.- vaccinated
mice had died. Intradermal vaccination produced the weakest protection
because only mice from this group died from challenge with 28 CFU SCHUS4.
The vaccination dose has no effect on the level of protection (Fig. 1).
iii. Intranasal LVS vaccination generated similar levels of protection against i.n.
SCHU S4 challenge in the out bred NIH-Swiss mice and inbred BALB/c mice
(Fig. 2).
iv. We will continue to monitor the survival for a total of 30 days. Additional deaths
over the next 3 weeks may affect our interpretation of the results.
b. Data is located in notebook 81, on pages 48-55
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
5
Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
4. Significant decisions made or pending
Wait 6 weeks after vaccination and confirm LVS clearance before SCHU S4 challenge.
5. Problems or concerns and strategies to address
NA
6. Deliverables completed
None
7. Quality of performance
Good
8. Percentage completed
6%
9. Work plan for upcoming month
a. Ftc 7: We are repeating experiment Ftc6 to determine the LD50 for the LVS stock and its
growth and clearance kinetics in NIH-Swiss mice and BALB/c mice following i.n., i.d., and
s.c. inoculation. We used the results from Ftc6 as a reference and included doses that
will enable us to calculate the LD50. The doses are: 5 x 103, 5 x 104 and 5 x 105 per
mouse for i.n. inoculation and 5 x 106 per mouse for i.d. and s.c. inoculation. The results
at this point are similar to those from Ftc6.
b. Ftc7B: We will repeat experiment Ftc6B to determine effect of vaccination dose and route
on the level of protection in mice. As discussed at the Charleston meeting, we will wait
for 6 weeks and confirm clearance of LVS before challenging them intranasally with
SCHU S4.
c. Ftc9: We are determining the i.n. LD50 of the SCHU S4 stock in NIH-Swiss mice. We
expect the LD100 to be very low and, therefore, we inoculated the mice with 10 1 , 102 and
103 CFU.
d. Rats: We are developing Standard Operating Procedures (SOPs) for working with rats.
These will include methods for infecting rats i.n., s.c. and i.d., monitoring their health
status, and processing the lungs, spleens, and livers to determine the bacterial load.
10. Anticipated travel
None
11. Upcoming Contract Authorization (COA) for subcontractors
None
Milestone 12
Milestone description: Assays for detecting relevant immune responses in animals & humans
developed
Institution: LBERI
1. Date started: 2/23/2006
2. Date completed: in progress
3. Work performed and progress including data and preliminary conclusions
Julie Wilder traveled to and prepared a presentation for the TVDC Kickoff meeting in Charleston,
SC, held 3/28-3/30; Dr. Wilder completed background reading pertinent to this presentation and
meeting.
4. Significant decisions made or pending
Ayako Monier was hired on 3/27 as Dr. Wilder’s technician and has been training on relevant
immunoassays in her laboratory at LBERI.
5. Problems or concerns and strategies to address
None
6. Deliverables completed
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
None
7. Quality of performance
Good
8. Percentage completed
0.1% of scientific work has been completed
9. Work plan for upcoming month
a. Ms. Monier will complete IACUC Animal Care training modules in order to be approved to
work with animals.
b. Ms. Monier will continue to train on relevant immunoassays in the laboratory.
c. Dr. Wilder and Ms. Monier will attend a meeting with Dr. Lyons and other UNM/LRRI
team members to discuss upcoming tasks.
10. Anticipated travel
None anticipated at the present time
11. Upcoming Contract Authorization (COA) for subcontractors
None for this milestone.
Milestone 34
Milestone description: Pilot studies for optimization of RNA isolation and hybridization
conditions done
Institution: UNM/ASU-Johnston
1. Date started: 3/01/06 UNM
2. Date completed: pending
3. Work performed and progress including data and preliminary conclusions
UNM prepared and sent SCHU S4 RNA (81 ug) and LVS RNA (61ug) and SCHU S4 DNA (35 ug)
and LVS DNA (31ug) to ASU. RNA demonstrated excellent integrity. Nucleic acids arrived at ASU
at correct temperature.
4. Significant decisions made or pending
None.
5. Problems or concerns and strategies to address
None
6. Deliverables completed
None
7. Quality of performance
Good
8. Percentage completed
0.5%
9. Work plan for upcoming months
ASU will utilize these LVS and SCHU S4 genomic DNAs and RNAs to test hybridization of printed
microarrays. ASU analyzes microarray gene expression data utilizing genomic DNA as
normalizing control. ASU utilizes cDNA made from bacterial RNA to analyze bacterial gene
expression by microarray data analysis.
10. Anticipated travel
Dr. Lyons and Barbara Griffith are traveling to ASU on 4/25/06 and will discuss scientific
strategies and plans.
11. Upcoming Contract Authorization (COA) for subcontractors
See MS#1 above
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
Milestone 41
Milestone description: Optimization of psoralen treatment and characterization of KBMA
F.novicida; Determine the amount of S-59 and UVA required to inactivate uvr mutants,
Determine extent of metabolic activity of uvr mutants after S-59 and UVA inactivation,
Determine the level of virulence attenuation of KBMA uvr strains in mice
Institution: Cerus
1. Date started: 3/2/06
2. Date completed: TBD
3. Work performed and progress including data and preliminary conclusions
Obtained current quotations for animal racks and cages needed for animal studies
Submitted application to USDA for permit to import and transport Francisella tularensis ssp.
novicida strains
Ordered media components to cultivate F. novicida
4. Significant decisions made or pending
None
5. Problems or concerns and strategies to address
Awaiting completion of subcontract with UTSA for uvr mutants to be completed and delivered to
Cerus.
6. Deliverables completed
None
7. Quality of performance
NA
8. Percentage completed
1%
9. Work plan for upcoming month
Obtain approval for and order equipment necessary for experiments.
Begin optimization of growth and inactivation conditions for U112 parental strain of F. novicida for
baseline characterization.
10. Anticipated travel
None
11. Upcoming Contract Authorization (COA) for subcontractors
None
Milestone 46
Milestone description: Scale up of KBMA LVS vaccine production; Optimize large–scale
Francisella tularensis culture conditions, Establish 3L culture scale purification conditions,
Optimize 3L scale photochemical inactivation process, Verify protective immunogenicity of
vaccine candidates produced by optimized large-scale process
Institution: Cerus
1. Date started: 3/10/06
2. Date completed: TBD
3. Work performed and progress including data and preliminary conclusions
Obtained current quotations for manufacturing equipment for 3L scale process development.
Submitted application to USDA for permit to import and transport LVS
4. Significant decisions made or pending
Standardized medium to be agreed upon by Working Group established at TVDT meeting in
Charleston, SC.
5. Problems or concerns and strategies to address
None
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Tularemia Vaccine Development Contract: Technical Report
Period: 3/01/2006 to 3/31/2006
Due Date: 4/14/2006 and Prepared by: Barbara Griffith, Terry Wu, Justin Skoble and Bob
Sherwood
6. Deliverables completed
None
7. Quality of performance
NA
8. Percentage completed
1%
9. Work plan for upcoming month
Fermentation equipment to be approved and ordered
10. Anticipated travel
None
11. Upcoming Contract Authorization (COA) for subcontractors
None
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