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Study of the corneal endothelial infection after Herpes simplex virus type 1 (HSV1) in a murine model: comparison between in situ confocal microscopy (ISCM) and immunofluorescent analysis on histological sections
Author(s) -
POGORZALEK N,
HUOT N,
CREPIN S,
FRANCELLE L,
OFFRET H,
NAAS T,
LABETOULLE M
Publication year - 2008
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2008.561.x
Subject(s) - confocal microscopy , art , confocal , humanities , physics , microbiology and biotechnology , biology , optics
Purpose To compare images obtained with ISCM of cornea and immunofluorescence analysis in a mouse model of HSV1 ocular infection (keratouveitis), with the aim of assessing inflammatory reaction (macrophages, lymphocytes and polymorphonuclear cells) and apoptosis during acute infection. Methods Fifty mice were analyzed using ISCM of left infected corneas, 6 days after HSV1 inoculation. The corneas were then flat mounted, and stained using immmunofluorescence for viral infection, inflammatory markers (macrophages, lymphocytes, PMNs) and apoptosis. Results ISCM showed multiple retrocorneal cellular precipitates on the inner face of the cornea, with a cluster distribution, and multiple hyper‐ and hypo‐reflective cells randomly distributed all over the endothelium. Histological analysis proved precipitates to be centered by infected endothelial cells and macrophages. Besides, hyper‐ and hypo‐reflective ISCM signals could correspond to either lymphocytes or PMNs, which were found on flat corneas to be scattered onto the endothelium cells, or to apoptotic cells. Most of them corresponded to lymphocytes closely localised near infected endothelial cells (ongoing study). Conclusion As ISCM images observed in our mouse model were very similar to those observed in human clinical practice with IVCM, our results provide precious information on the biological meaning of clinical observations. In the future, IVCM could be routinely used to monitor herpetic infection in pre‐clinical studies, for example in experiments assessing the efficacy of new antiviral strategies.