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C. Clement, V. Tiwari, B.Y. Yue, D. Shukla; Susceptibility of Human Corneal Fibroblasts to Herpes Simplex Virus Type 1: An Electron Microscopy Study . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4629.
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Purpose: To investigate the entry of herpes simplex virus 1 (HSV-1) into human corneal fibroblasts using electron microscopy. Methods: This study was designed to assess the entry of herpes simplex virus 1 in cultured corneal fibroblasts, especially the anatomical changes in cell structure as a result of viral attachment and fusion using transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Cultures of corneal fibroblasts were derived from stroma of donor eyes. A recombinant strain of wild type HSV-1 (KOS gL86) was purified by sucrose gradient centrifugation and used for infecting the cultured corneal fibroblasts. A time course study of viral entry was performed using both TEM and SEM. Results: Our results show that HSV-1 attachment and fusion with human corneal fibroblasts is characteristically unique in comparison to cell lines like transformed Chinese hamster ovary (CHO-K1) cells and naturally susceptible HeLa cells. Although all the cell lines allowed viral entry, the corneal fibroblasts showed initial resistance by allowing delayed entry. Critical examination of micrographs revealed a slower attachment of viral particles. It was not totally unexpected because published data suggest corneal fibroblasts produce very little amount of heparan sulfate which the virus needs for attachment. The points of attachment of viruses on the corneal fibroblasts surfaces revealed that these sites were indented, this feature was absent in the nonprimary cells, suggesting some pressure from the virus initiating entry. Also, unlike the other cell types where viruses were detected on the nuclear membrane within a few minutes, it took more than one hour to detect viral particles in the nucleus of corneal fibroblasts. Conclusions: We have demonstrated the susceptibility of human corneal fibroblasts to HSV-1 entry, suggesting that these cells widely express HSV-1 receptors like HVEM, Nectin-1 and Nectin-2. By their nature these primary cells may provide a more useful model for studying virus cell interactions in ocular herpes infection.
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