Tularemia Vaccine Development Contract: Technical Report
Period: 2/01/2006 to 2/28/2006
Due Date: 3/14/2006 and Prepared by: Barbara Griffith and Terry Wu
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, 5
Inactive milestones: 3, 4, 6-30, 31-54
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, and Cerus have signed contracts with COA number assigned. LBERI has
COA#2 as of 2/23/06, ASU has COA#3 as of 3/2/06, and Cerus has COA#4 as of 3/2/06
UTSA- subcontract is in negotiation, with teleconference meeting on 3/10/06
The timeline report will be in MS Project.
ASU, UTSA, Cerus and LBERI have fully signed Datasharing plans with UNM.
UNM, ASU, UTSA, Cerus and LBERI have NDAs signed with CSC/DVC
NDAs- UNM with DVC subcontractors
i. NDAs executed : Umea University and NRC
ii. NDAs pending: DSTL
g. . . Strategic Work Plan: has been provided and revisions will be given to Contract Officer
and Project Officer by 3/15/06
b.
c.
d.
e.
f.
4. Significant decisions made or pending
a. Barbara is preparing the customized timeline report template in MS Project and the
expense report template by milestone is in preparation by Mindy Tyson and Michelle
Matteucci
b. Final fully executed subcontracts will be submitted to Ross Kelley within 30 days of the
COA number assignment.
5. Problems or concerns and strategies to address
a. UTSA- UNM negotiating with their first round of language requests on 3/10/06 in
teleconference, with UTSA legal representatives and Noe Saldana (Pre-award for UTSA)
6. Deliverables completed
None- subcontracts will be finalized when fully executed and returned to Ross Kelley.
7. Quality of performance
Good
8. Percentage completed
80%
9. Work plan for upcoming month
UNM to complete the revised Strategic Work Plan by 3/21/06
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Tularemia Vaccine Development Contract: Technical Report
Period: 2/01/2006 to 2/28/2006
Due Date: 3/14/2006 and Prepared by: Barbara Griffith and Terry Wu
10. Anticipated travel
a. Ten members of the UNM TVDC team will travel to Charleston, SC for a joint DVC and
UNM TVDC meeting on 3/28/06 to 3/30/06
b. With two nights lodging per person, per diem, ground transportation, round trip airfare, and
meeting room costs, UNM anticipates a cost of $13,500.
c. Rick Lyons and Barbara Griffith will travel to Cerus site on 3/22/2006 for one day 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 #1- 1/17/06- approves purchase of 3 computers, which were budgeted in ASU’s
b.
c.
d.
e.
f.
g.
subcontract but which were not listed in Government Property Schedule IA
COA#2- 2/23/06- approves subcontract with LBERI
COA#3- 3/2/06 approves subcontract with ASU
COA#4- 3/2/06 approves subcontract with Cerus
COA pending- potential purchase of equipment for monitoring blood coagulation
parameters in the ABSL facility and the complete bronchscopy system for LBERI usage.
COA pending- for biobubble purchase for LBERI installation
Request to Ross submitted to correct the 4 cost errors in the Government Property
Schedule 1A
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.
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. At an approximate cost of $3380 to $10,000, 30 people cost
$101,400 minimum and 51 people cost $172,380 minimum. UNM is having discussions
with LBERI regarding covering 20 of their 41 people. Barbara compared the prioritized
vaccination list supplied by Chuck Hobbs (attached) with the personnel named in the
UNM business proposal and there are 13 overlaps. All prioritized 41 LBERI scientists
and staff fulfill roles and responsibilities that will be utilized by the TVD Contract. The
24hr/7 day year round care of the non-human primates requires 3 shifts of coverage for
such responsibilities as necropsy, animal care, veterinary care, pathology, and more.
d. A request for a COA for the biobubble for LBERI has been submitted to Ross Kelley
5. Problems or concerns and strategies to address
a. Barbara is preparing a revised budget for the “USAMRIID” expense category, which has
been retitled “Occupational Health” .The new budget will cover travel expenses for 20
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Tularemia Vaccine Development Contract: Technical Report
Period: 2/01/2006 to 2/28/2006
Due Date: 3/14/2006 and Prepared by: Barbara Griffith and Terry Wu
LBERI staff, for 6 from UTSA and for 4 from UNM. It will also cover pre-health screening
expenses for the LVS vaccinations, which are required by USAMRIID. It will also cover
the cost of the biobubble
6. Deliverables completed
None
7. None Quality of performance
Good
8. Percentage completed
10%
9. Work plan for upcoming month
a. 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.
b. Revise the budget for Occupational Health, to cover travel and prehealth screening for
30-31 scientists and staff.
10. Anticipated travel
None in the next month; travel could occur in any month between April and June 2006
11. Upcoming Contract Authorization (COA) for subcontractors
a. COA- has been submitted to request the purchase of a Biobubble ($20-24,000) as
additional “Occupational Health” personal protective equipment at LBERI
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 have completed the first in vivo experiment using the LVS stock from DVC (Experiment
code Ftc6) and data is located in Notebook 81 on pages 36 to 45.
b. The purpose of this experiment was to: (1) Determine the LD50 for this LVS stock in out
bred NIH Swiss mice when given i.n., s.c. or i.d.; (2) Determine the kinetics of bacterial
growth associated with each inoculation route; and (3) Compare the sensitivity of out bred
NIH Swiss mice and inbred BALB/c mice to i.n. LVS infection.
c. Based on preliminary results derived with the LVS stock currently used at UNM, NIH Swiss
mice were infected s.c. or i.d. with 5 x 104 to 5 x 106 CFU/mouse and i.n. with 5 x 103 to 5 x
106 CFU/mouse. We did not expect any of the infected NIH Swiss mice to die. As shown in
Fig. 1A, mice infected s.c. or i.d. survived infection with the highest possible dose we could
achieve with DVC’s stock. However, mice infected i.n. only survived the lowest dose of 1.7 x
103CFU/ml, Similar results were obtained following i.n infection of BALB/c mice (Fig. 1B).
Thus, the LVS stock from DVC has similar virulence in out bred Swiss mice and in inbred
BALB/c mice.
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Tularemia Vaccine Development Contract: Technical Report
Period: 2/01/2006 to 2/28/2006
Due Date: 3/14/2006 and Prepared by: Barbara Griffith and Terry Wu
NIH Swiss
A
Intranasal
Subcutaneous
1.8 x 10 6
1.3 x 10 5
1.8 x 10 4
1.7 x 10 3
50
25
0
100
75
6
2.6 x 10
2.6 x 10 5
2.6 x 10 4
50
25
0
0
10
20
30
B
10
20
30
Intranasal
100
75
1.8 x 10 4
1.7 x 10 3
2.4 x 10 2
2.1 x 10 1
50
25
2.6 x 10 6
2.6 x 10 5
2.6 x 10 4
50
25
0
Days
BALB/c
75
0
0
Days
Percent survival
Percent survival
100
75
Percent survival
Percent survival
100
Intradermal
10
20
30
Days
Figure 1. Survival of mice infected with LVS.
Groups of 10 NIH Swiss mice (A) and
BALB/c mice (B) were infected i.n., s.c., or id
with the indicated dose of LVS and monitored
for survival.
0
0
10
20
30
Days
d. To determine the kinetics of bacterial growth, twelve additional mice were infected with the
highest dose of each route. Table 1 shows that 4 days after i.n. infection with 1.8 x 106 CFU,
LVS had proliferated 1-2 log10 in the lung and disseminated to the spleen and liver (Table 1).
Results from later time points could not be obtained because most of the mice died within 7
to 10 days of infection,
e. The kinetics of bacterial growth in mice infected s.c. or i.d. were very similar. Four days
after infection with approximately 2.5 x 106 CFU, only a small fraction of the inocula had
disseminated systemically and most of those preferentially colonized the spleen and liver
(Table 2 and 3). By d 10 and d 15, most of the mice had cleared the infection.
Table 1. Kinetics of bacterial growth in NIH Swiss mice after i.n. infection
Day of
Tissue infection
Mouse
1
2
3
4
Lung
0
2.4 x 106 1.3 x 106 1.7 x 106
4
1.8 x 107 1.6 x 108 8.4 x 107 6.8 x 107
Spleen
4
2.2 x 105
1.1 x 106
6.5 x 105
1.5 x 106
Liver
4
9.1 x 105
2.0 x 106
6.9 x 105
2.8 x 106
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Tularemia Vaccine Development Contract: Technical Report
Period: 2/01/2006 to 2/28/2006
Due Date: 3/14/2006 and Prepared by: Barbara Griffith and Terry Wu
Table 2. Kinetics of bacterial growth in NIH Swiss mice after i.d. infectiona
Day of
Tissue infection
Mouse
1
2
3
4
Lung
4
2.0 x 101 2.0 x 101 1.2 x 106 2.8 x 102
10
4.0 x 101
NDb
ND
ND
15
ND
ND
ND
ND
Spleen
3.4 x 103
2.0 x 101
ND
4
10
15
1.4 x 103
2.0 x 101
ND
4.5 x 106
ND
4.8 x 101
3.8 x 103
ND
ND
4
3.8 x 103 4.4 x 102 5.2 x 107 1.1 x 103
10
ND
ND
ND
ND
15
ND
ND
ND
ND
a Mice were infected i.d. with approximately 2.5 x 10 6 LVS
b ND = not detected. Limit of detection for this assay is 20 CFU/organ
Liver
Table 3. Kinetics of bacterial growth in NIH Swiss mice after s.c. infectiona
Day of
Tissue infection
Mouse
1
2
3
4
Lung
4 2.0 x 101 2.0 x 101 1.4 x 102 2.0 x 101
10
ND
ND
ND
ND
15
ND
ND
3.0 x 101
ND
Spleen
4
10
15
2.9 x 103
6.0 x 101
ND
1.7 x 104
ND
ND
4.0 x 104
ND
ND
2.8 x 103
8.0 x 101
ND
4 7.6 x 102 5.2 x 103 4.5 x 104 3.6 x 103
10 1.6 x 102
ND
ND
ND
15
ND
ND
ND
ND
a Mice were infected s.c. with approximately 2.9 x 10 6 LVS
b ND = not detected. Limit of detection for this assay is 20 CFU/organ
Liver
4. Significant decisions made or pending
a. Experiment Ftc6 with mice will be repeated as experiment Ftc7
b. We decided to purchase Fischer 344 rats from NCI-Frederick for the next phase of our
study because its price is lower than all other vendors. We will set up a standing order to
ensure delivery of our requested number of rats
5. Problems or concerns and strategies to address
NA
6. Deliverables completed
None
7. Quality of performance
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Tularemia Vaccine Development Contract: Technical Report
Period: 2/01/2006 to 2/28/2006
Due Date: 3/14/2006 and Prepared by: Barbara Griffith and Terry Wu
Good
8. Percentage completed
4%
9. Work plan for upcoming month
a. We will repeat the Ftc 6 experiment in Ftc 7 to determine the LD50 for DVC’s LVS stock
and the LVS growth kinetics in NIH Swiss mice and BALB/c mice following i.n., i.d., and
s.c. infection. We will use the results from Ftc6 as a reference to determine the
appropriate dose range for infection in order to accomplish the stated objective.
b. UNM has received 100 vials of SCHU S4 from Fisher Biosciences on March 9, 2006. We
will determine the titer of this stock and the LD50 in NIH Swiss mice after i.n. infection.
The mice that had survived LVS infection in Ftc6 have essentially been vaccinated and
will therefore be challenged to determine the whether LVS vaccination induces immunity
against i.n. SCHU S4 challenge.
10. Anticipated travel
a. Dr. Terry Wu will be attending the TVD Contract meeting in Charleston, SC in Mar 2006.
11. Upcoming Contract Authorization (COA) for subcontractors
None
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