1 Cytokine Expression in the Central Nervous System and Periphery during Infection with Rabies Virus Nicholas Johnson Rabies and Wildlife Zoonoses Group WHO Collaborating Centre for the Characterization of Rabies and Rabies-Related Viruses
Mar 27, 2015
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Cytokine Expression in the Central Nervous System and Periphery during Infection with Rabies Virus
Nicholas JohnsonRabies and Wildlife Zoonoses Group
WHO Collaborating Centre for the Characterization of Rabies and Rabies-Related Viruses
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Interferon induction following infection with a fixed strain of Rabies virus (Challenge Virus Standard)
RV nucleoprotein
Interferon-
Interferon –
Interferon-
603bp
x RV negative RV positive
310bp
603bp
310bp
310bp
310bp
-actin
TNF-
Johnson et al., 2006 JGV 87, 2663
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Objectives
Measure inflammatory changes in response to infection with a non-fixed strain
Observe inflammation within the Central Nervous System
Identify possible mediators of inflammation (cytokines / chemokines)
Compare the CNS to the periphery during infection
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The Model
00 2 4 6 8 10 12
Inoculate Antibodies Signs Death
Isolate RV 61 (UK human, ex-India)
CD1 outbred mice
Subdermal inoculation / hind footpad
11d course (sampled at 0, 5, 11)
Brain / salivary glands: Transcript
Spinal cord: Histopathology
Blood: Cytokine / Antibody
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Anti-RABV immunostaining in the Dorsal Root Ganglion & Spinal Cord
Dorsal Root Ganglion (d11) Spinal cord (d11)
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Inflammatory changes observed in the DRG and Spinal Cord
DRG (Media only) DRG (RABV d11)
Spinal cord (Media only) Spinal cord (RABV d11)
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T- and B-Cell Lymphocyte Immunostaining
CD45R/220
CD3
Dorsal Root Ganglia Spinal cord
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Inflammatory Cytokine Detection in the Brain
CCL3
M
RABV - RABV + Day O Day 5 Day 11
GAPDH
CCL2
CCL4
CCL5
Overt disease
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Chemokine transcript response in the brain
0
30
60
90
120
0 5 110
100
200
300
400
0 5 11
0
100
200
300
400
0 5 11
0
80
160
240
320
0 5 11
0
600
1200
1800
2400
0 5 110
1200
2400
3600
4800
0 5 11
INF- CCL2 (MCP-1) CCL3 (MIP-1)
CCL4 (MIP 1) CCL5 (Rantes) CXCL10 (IP10)
Tra
nscr
ipti
on f
old
incr
ease
Time post inoculation (days)
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Peripheral Changes
Salivary glands: No change in TLR-3 transcripts.Increases observed in levels of a number of inflammatory transcripts (IL-6, CCL5, CXCL10, 2’-5’ OAS1)
Serum:
Neutralising antibodies detected as early as day 5.No change in a range of cytokines (INF-, IL-2, IL-6, IL-10, TNF-)
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Conclusions
Inflammatory changes are observed in the CNS during neurovirulent infection with RABV.
Infiltrating lymphocytes are predominantly CD3+ (but not observed in the brain)
Interferons and chemokines are up-regulated in response to infection.
Chemokine responses proceed detection of RABV in the brain.
Chemokines that attract T cells show the greatest up-regulation.
These changes are not reflected in the periphery.
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Acknowledgements
Co-authors
Veterinary Laboratories Agency
Anthony FooksSharon BrookesKaren MansfieldDerek HealyDaniel HicksAlex Nunez
University of Edinburgh
John FazakerleyClive McKimmie
Funding
Defra (UK): grant SE0524