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Zinkernagel, M.S., Chinnery, H.R., Petitjean, C., Wikstrom, M.E., McMenamin, P.G. and
Degli-Esposti, M.A. 2009, In vivo imaging of mCMV infection in the eye, in ARVO 2009:
Proceedings of the 2009 Association for Research in Vision and Opthalmology Conference,
ARVO, Fort Lauderdale, Flo., pp. 1-1.
Available from Deakin Research Online:
http://hdl.handle.net/10536/DRO/DU:30052114
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Copyright: 2009, Association for Research in Vision and Ophthalmology.
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Presentation Abstract
Program#/Poster#: 2388/D695
Abstract Title:
In vivo Imaging of mCMV Infection in the Eye
Presentation
Start/End Time:
Monday, May 04, 2009, 3:45 PM - 5:30 PM
Location:
Hall B/C
Reviewing Code:
419 viral infections: intraocular - IM
Author Block:
M.S. Zinkernagel1, H.R. Chinnery2, C. Petitjean1, M.E. Wikstrom1, P.G. McMenamin2,3, M.A. Degli
-Esposti1. 1Experimental Immunology, Centre for Ophthalmology and Visual Sciences (COVS),
Lions Eye Institute, University of Western Australia, Perth, Australia; 2Ocular Immunology, Centre
for Ophthalmology and Visual Sciences (COVS), University of Western Australia, Lions Eye
Institute, Perth, Australia; 3School of Anatomy and Human Biology, University of Western
Australia, Perth, Australia.
Keywords:
701 retinochoroiditis, 558 inflammation, 552 imaging/image analysis: non-clinical
Abstract Body:
Purpose: To develop a technique by which murine cytomegalovirus (mCMV) can be tracked and
monitored in various ocular compartments in vivo, and to investigate the dynamics and kinetics of
ocular mCMV infection.
Methods: The development of a recombinant mCMV tagged with enhanced green fluorescent
protein (eGFP-MCMV) has made in vivo imaging of this pathogen feasible. Immunocompetent
BALB/c mice were infected intravitreally with eGFP-MCMV. On sequential days after inoculation,
infection in the eye was monitored with a Heidelberg retinal angiograph (HRA). Eyes were also
processed for histology and immunfluorescence microscopy to determine the nature of infected
cells in various ocular compartments. At indicated times after infection, FACS analysis was
performed to investigate recruitment of T cells into the retina.
Results: Green fluorescent signal appeared 24 hours after intraocular injection, with scattered foci
visible around the posterior pole of the retina. eGFP levels in the retina reached a maximum
between days 6 and 12, and signal was commonly found to accumulate around the optic nerve
head. At day 25 the signal became weaker, suggesting a decrease in the frequency of infected cells
in the retina. Signal in the iris was observed from day 4 and lasted up to day 50, with signal
commonly seen on the anterior curvature of the lens. Examinations of retinal and iris tissue
wholemounts by immunofluorescence revealed signal localized in the inner retina, iris stroma and
on the anterior lens capsule. FACS analysis of retinal tissue showed a recruitment of both CD8+
and CD4+ T cells from day 6 post infection. Tetramer analysis revealed a high frequency of
antigen-specific CD8+ T cells accumulating in the retina on day 12.
Conclusion: The ability to noninvasively monitor infectious agents in the eye may improve our
knowledge of the course and pathogenesis of intraocular infections and could lead to further
clarification of the mechanisms by which the immune system responds to intra-ocular pathogens.
Commercial
Relationships:
M.S. Zinkernagel, None; H.R. Chinnery, None; C. Petitjean, None; M.E. Wikstrom,
None; P.G. McMenamin, None; M.A. Degli-Esposti, None.
Support:
Swiss National Science Foundation, Grant No PBZHB-121040 and Lions Eye Institute, Perth,
Western Australia
©2009, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to www.iovs.org to access the
version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
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