BRM's Capabilities in Virology, Viral Immunology and Pathology: HSV-1, HSV-2, Influenza, RSV, LCMV, and CMV. For more information, please visit our website at www.BRMCRO.com.

1. Biomedical Research Models, Inc.
Contract Discovery Research
Virology, Viral Immunology and Pathology

2. BRM Capabilities
in Infectious Diseases
• NIH/NIAID, CDC funded mucosal HSV-2 and
RSV vaccine research grants ($6.8M)
• NIAID has awarded a seven year contract to
develop animal models of infectious diseases to a
research consortium including:
◦ BRM
◦ University of Massachusetts Medical School
◦ US Army Medical Research Institute of Infectious
Diseases
◦ Jackson Laboratories

3. In Vivo
• Virus Infection Models
• Vaccines
• Antiviral
In Vitro
• Immunology
(B, T cell and mucosal
responses),
Virology
(Plaque assay, RT-PCR)
and Pathology

4. Viral Infection Models
• HSV-2
◦ Female mice and guinea pigs (female and male)
• HSV-1
◦ Mouse ocular infection model and lung infection
• Influenza virus
◦ Mouse adapted human infection model and others
• Respiratory syncytial virus (RSV, mice and cotton rats)
• Lymphocytic choriomeningitis virus (LCMV) and
Cytomegalovirus (CMV)
◦ Chronic and Acute infection in Mice
• Humanized mouse models for infectious diseases
◦ NIH has awarded a seven year contract to a research
consortium

5. Influenza Virus Infection Model
• Intranasal instillation of virus to monitor mortality,
morbidity (body weight loss, decreased activity, less
food and water intake, show tendencies to huddle,
increased respiration, ruffled fur is evident)
• Test protective and therapeutic efficacy of vaccines,
immunomodulation agents and antivirals
• Evaluate immune responses, protection and pathology

6. Survival Post Influenza Infection
(mice, n=5/group)
Percentage of Survival
100%
80%
60%
40%
10 µg Flu Vaccine
1 µg Flu Vaccine
20%
0.1 µg Flu Vaccine
naïve
0%
D0
D1
D2
D3
D4
D5 D6 D7 D8
Days Post Infection
D9
D10 D11 D12 D13 D14

7. Body Weight Post Influenza Infection
(mice, n=5/group)
Relative weight change post influenza
challenge
115
(10 mg vaccine)
Relative weight change post influenza
challenge
120
(0.1 mg vaccine)
Relative weight
Relative weight
110
110
105
100
100
90
80
70
95
60
0
2
4
6
8
0
Days post infection
2
4
6
8
Days post infection
110
Relative weight
Relative weight change post influenza
challenge
120
(Naive )
110
Relative weight
Relative weight change post influenza
challenge
120
(1 mg vaccine)
100
90
100
90
80
80
70
70
60
0
2
4
Days post infection
6
8
0
2
4
Days post infection
6
8

8. HSV Infection Models
• Intravaginal inoculation of HSV-2 virus to monitor
mortality, morbidity (clinical score, body weight loss) and viral
load during primary and recurrent phases for both female mice
and guinea pigs.
• HSV-2 infection model in male guinea pig
• Intranasal infection with attenuated and virulent HSV-1 strains
• Test systemic and mucosal immune responses and protection
• Investigate latency and recurrence using clinical scores and RTqPCR methods

9. HSV-2 Clinical Score Grading System
•
•
•
•
•
0 – No apparent clinical signs
1 – Redness or mild swelling
2 – Raw patch, blister, moderate swelling
3 – Multiple blisters or raw patches
4 – Large blisters with severe and/or deep
sores; and/or urinary retention; and/or hind
limb paralysis; animal euthanized
• 5 – Animal found dead

10. Survival Post HSV-2 Infection
(mice, n=5/group)
100
Percent of Survival
80
60
40
Naïve
Vaccine 1
20
Vaccine 2
Vaccine 3
0
0
5
10
Days Post Infection
15
20

11. Body Weights Post HSV-2 Infection
(mice, n-5/group)
103
Percent of Body Weight
101
99
97
95
93
91
naïve
89
Vaccine 1
87
Vaccine 2
Vaccine 3
85
0
1
2
3
4
5
6
7
8
9
10
11
12
Days Post Infection
13
14
15
16
17
18
19
20

12. HSV-1 Ocular Infection Model
Acclimation of mice
(4-7 days)
Day 1: Corneal delivery of
compound(s)
Day 1: Corneal Infection of
HSV-1 viruses
Daily Clinical Assessment:
Observe Facial lesions and
swelling (3 weeks)
Humane Endpoints
Collection of fluid from both
eyes for testing viral
shedding
(Days 0 to 7, 14, 21)
Organ and Tissue Collection

13. HSV-1 Clinical Readouts
• HSV-1 KOS strain
• Daily Clinical Scoring
◦ Facial swelling based on a grading scale of 0-6
◦ Bilateral facial lesions on a grading scale of 0-8
(usually initiating after day 6)
• Virus titration of lacrimal drainage
• Histopathology of eyes and brain and other
organs of interest
◦ Stained with Luxol fast blue-periodic acidSchiff reagent and hematoxylin

14. Respiratory Syncytial Virus
(RSV)
• RSV virus A2 strain (High titer, quality stock)
• Infection of Balb/c mice results in low and
moderate level of replication that peaks on day4.
• Clinical symptoms: weight loss, changes in lung
function, ruffled fur
• Pulmonary eosinophilia

15. Acute and Chronic LCMV
Infection Models
• Mouse LCMV infection has been widely used to evaluate
therapeutic treatments against human viral infections
◦ Viral clearance or persistent infection
• Acute infection (Armstrong strain)
◦ Robust anti-viral T cell response
◦ Mediated by CD8+ virus-specific T cells
◦ Cleared within 2 weeks post infection
• Chronic infection (clone 13)
◦ Associated with functional impairment, exhaustion,
and deletion of virus-specific CD8+ T cells
◦ Lasting several months

16. CD8+ T Cell Responses 29 Days
Post LCMV Infection
TNF-a
GP 276
IFN-g
TNF-a
GP 276
IFN-g
LCMV Clone 13
NP 396
NP 396
LCMV Armstrong
CD44
CD44
CD44

17. The Establishment of Chronic Infection
in LCMV clone 13 Infected Mice
Viral titers in the serum post LCMV
infection
Viral titers in Kidney and Lung
(Day 29 post infection)
5.00
5.00
4.50
4.00
Day 2
Day 8
3.50
Day 15
Day 22
3.00
Day 29
Viral titer (PFU/mL)
Viral titer (PFU/mL)
4.50
4.00
3.50
3.00
Kidney
Lung
2.50
2.00
2.50
1.50
2.00
1.00
LCMV Armstrong
LCMV Clone 13
LCMV Armstrong
LCMV Clone 13

18. Humanized Mouse Models
• Under the aegis of the NIAID contract, the
following humanized mouse strains are being
evaluated:
◦
◦
◦
◦
◦
NODscidIL2rgnull mice
HLA-A2 transgenic NOD-scid IL2rnull mice
Human Blys transgenic NOD-scid IL2rnull mice
TLR4KO NOD-scid IL2rnull mice,
Myd88 KO NOD-scid IL2rnull mice

19. Cytomegalovirus
(CMV)
• Infection is species-specific for HCMV, MCMV, RCMV and
guinea pig CMV
• Serious diseases in immunosuppressed patients ( 50-60% organ
and bone marrow transplantation patients)
• Primary infection is followed by persistent and recurrent
infections
• Severe combined immunodeficient (SCID) mice, CB-17 SCID
mice which lack of T and B cells are sensitive to MCMV
infection i.p.
(blood, liver, spleen, lung, adrenal, pancreas, kidney, brain, sali
vary)
• Infection of Balb/c mice (3-week-old) i.p. , dose-dependent
mortality
• SCID-hu mice for ocular HCMV infection

20. Summary
• Scientific team with decades of combined
experience
• Tailored study design for your project needs
• Competitive pricing
• Flexible and rapid study start to
your timelines