April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
In vivo Imaging of mCMV Infection in the Eye
Author Affiliations & Notes
  • M. S. Zinkernagel
    Experimental Immunology, Centre for Ophthalmology and Visual Sciences (COVS), Lions Eye Institute, University of Western Australia, Perth, Australia
  • H. R. Chinnery
    Ocular Immunology, Centre for Ophthalmology and Visual Sciences (COVS), University of Western Australia, Lions Eye Institute, Perth, Australia
  • C. Petitjean
    Experimental Immunology, Centre for Ophthalmology and Visual Sciences (COVS), Lions Eye Institute, University of Western Australia, Perth, Australia
  • M. E. Wikstrom
    Experimental Immunology, Centre for Ophthalmology and Visual Sciences (COVS), Lions Eye Institute, University of Western Australia, Perth, Australia
  • P. G. McMenamin
    Ocular Immunology, Centre for Ophthalmology and Visual Sciences (COVS), University of Western Australia, Lions Eye Institute, Perth, Australia
    School of Anatomy and Human Biology, University of Western Australia, Perth, Australia
  • M. A. Degli-Esposti
    Experimental Immunology, Centre for Ophthalmology and Visual Sciences (COVS), Lions Eye Institute, University of Western Australia, Perth, Australia
  • Footnotes
    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.
  • Footnotes
    Support  Swiss National Science Foundation, Grant No PBZHB-121040 and Lions Eye Institute, Perth, Western Australia
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 2388. doi:
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      M. S. Zinkernagel, H. R. Chinnery, C. Petitjean, M. E. Wikstrom, P. G. McMenamin, M. A. Degli-Esposti; In vivo Imaging of mCMV Infection in the Eye. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2388.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

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.

Keywords: retinochoroiditis • inflammation • imaging/image analysis: non-clinical 
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