Double Trouble:

Double Trouble:
Consequences of Immune Cell
Mis-Recruitment following Multiple
Viral Challenges
Christine Matullo
Glenn Rall
Research and Technology Symposium
20 May 2008

Overview
Question
Model
Outcomes
Significance
Opportunity
Lunch

Can peripheral viral infections-or
the immune responses to those infections-contribute
to CNS disease?
CNS diseases of
unknown etiology
-Multiple sclerosis
-Autism
-Alzheimer’s
-Lou Gehrig’s disease/ALS
-Parkinson’s
Each possess “hallmarks” of
infection:
-Chronic inflammation
-Relapsing/remitting
-Timing
…but no viruses have been
detected at the sites of damage

Damage
Virus
Putative roles for viruses in CNS disease
periphery
brain
Immune
Response
Co-localized Spatially distant
Damage
Virus

Outline
Question
Model
Outcome
Significance
Opportunity

Two-Virus Model
Recruitment Signal:
Measles Virus
Peripheral Challenge:
Lymphocytic
Choriomeningitis Virus

LCMV-Armstrong
1000-100,000 PFU
LCMV: Peripheral Challenge
6-8 d
10-15 d
Robust infection of peripheral
tissues
NO CNS INVOLVEMENT
Virus resolved;
Complete survival;
Lifelong protection
Oldstone et al, 1983

Model of neuronal measles virus infection
NSE-CD46 + transgenic mice
CD46:
One of the 2 identified
human MV receptors
NSE-CD46 + transgenic mice
receptor expressed only in neurons
In vivo
In vitro
Primary CD46 + neurons
Rall, et al, 1997

Immune-Mediated Protection of MV-infected
NSE-CD46 Transgenic Mice
% Survival
100
90
80
70
60
50
40
30
20
10
0
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Days Post-Infection
Immunocompetent adults
CD46/RAG2 -/- adults
NO PERIPHERAL INVOLVEMENT
Lawrence, et al, 1999

Route Site
i.c./i.n. CNS
i.p.
periphery
Immune
Profile
CNS:
CD4, CD4,
CD8
Periphery:
CNS:
none
Periphery:
CD8 > CD4
CD8
* Peak immune response for both viral infections: 6-10 dpi
none
Outcome
100%
Survival
100%
Survival
Patterson et al, 2001
Khanolkar, Fuller, & Zajac, 2002

Immune response
tissue-restricted
?
Immune response
ŅhomogenizedÓ

Outline
Question
Model
Outcome
Significance
Opportunity

Fraction Original Body Weight
Pathogenesis in doubly infected mice associated
with extensive weight loss
Days Post Infection
-Dose level independent
Uninf. (12/12)
+MV ic (13/13)
-Neuropathology is independent of any viral variable (e.g.,
tropism, clearance rate, virus level)
+LCMV ip (14/14)
+MV ic +LCMV ip
asymptomatic (12/38)
+MV ic +LCMV ip
symptomatic (26/38)

α-CD8 CD8 α-CD4 CD4
Increased CD8 + T-cell infiltration into
brains of doubly infected mice
+ MV
+ LCMV
Quantitation?
+ MV + LCMV

10-fold increase in CD8+ T-cell infiltration
into brains of doubly infected mice
Percent CD8+
CD4
Total CD8+ count
uninfected
10 5 2.24 0.082
10 4
10 3
10 2
10 1
CD8
95.3
10 1
10 2
10 3
10 4
2.38
10 5
+MV ic +MV ic +LCMV ip
5
10 19.6 0.21
10 4
10 3
10 2
10 1
64.4
10 1
10 2
10 3
5
10 5.7 0.39
2.2 % 14.7 % 56.7 %
469 14,050 170,518
n=15 n=28 n=37
10 4
15.8
10 5
10 4
10 3
10 2
10 1
32.6
10 1
ρ < 0.0001
About 60% of these CNS-infiltrating T cells are of LCMV specificity!
10 2
10 3
10 4
61.3
10 5

Is this unique to this model system, or can other pathogenic
combinations result in a similar outcome?
MV
Ectromelia virus
CD4
10
10 4
10 3
10 2
10 1
5 4.55 0.082
CD8
76
10 1
+ECTV +MV +ECTV
10 2
10 3
10 4
19.4
10 5
10 5 9.85 0.12
10 4
10 3
10 2
10 1
33.2
10 1
10 2
10 3
10 4
56.8
10 5

Poliovirus, IC
(using PVR transgenic mice)
LCMV
α-CD8
+PV ic +PV ic +LCMV ip

Outline
Question
Model
Outcome
Significance
Opportunity

I. Direct link to human disease:
midbrain herniation
--Occurs in humans following meningitis/encephalitis, brain tumor, trauma
(that is: increased intracranial pressure)--very precipitous death,
often of unknown cause
--As a result of edema, midbrain pushed through foramen into spinal
cord
--Because the origin of the optic nerves connects to the basal midbrain,
sudden, unilateral pupillary dilation is the key clinical indicator of
this condition

Pupillary Dilation and Edema
MV alone MV and LCMV
% Water Weight
uninfected
+LCMV ip
+MV ic
+LCMV ic
+MV ic
+LCMV ip
Sick
Healthy

I. Direct link to human disease:
midbrain herniation
--Occurs in humans following meningitis/encephalitis, brain tumor, trauma
(that is: increased intracranial pressure)--very precipitous death,
often of unknown cause
--As a result of edema, midbrain pushed through foramen into spinal
cord
--Because the origin of the optic nerves connects to the basal midbrain,
sudden, unilateral pupillary dilation is the key clinical indicator of
this condition
--A sense of the frequency:
Cerebral malaria: 300-500 million cases annually worldwide
>1 million deaths
50-75% of these (mostly children) show signs of midbrain herniation

II. Broadening how we think about
human CNS disease
CNS
DISEASE
Potential role for peripherally triggered immune responses in CNS diseases
with inflammatory component:
Brain tumors, MS, ALS, Parkinson’s, Alzheimer’s, stroke

Outline
Question
Model
Outcome
Significance
Opportunity

I. In theory, ANY immune stimulus could
contribute to immune cell mis-recruitment
Viruses
Bacteria
Parasites
Allergens
Autoimmune diseases
Vaccinations
Graft rejection
Tumor
Is there a protective/amelioritive role for anti-inflammatory drugs in CNS
diseases?
Development of complex animal models to understand complex human
diseases

II. Patient to patient variability
CNS DISEASE
~50% ~50%

Former:
Diane Lawrence
Catherine Patterson
Srdjan Askovic
Mindy Vaughn
Alec Belman
Eric Callahan
Michael Birnbaum
Jaimy Joy
Nina Makhortova
Lisa Gechman
Thanks…
Current:
Christine Matullo
Wes Rose
Ginger Young
Lauren O’Donnell
Jazz Skipworth
Anna Vorobyeva
Kevin O’Regan
Steve Conway
Support:
NINDS
NIAID
F.M. Kirby Foundation
Autism Speaks
Collaborators:
John Wherry, Wistar
Mark Curtis, TJU
Fox Chase:
-LAF
-FACS facility
-qRT-PCR facility
-MRI facility