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Bahram Bodaghi, Marlea E. P. Slobbe–vanDrunen, Andrzei Topilko, Emmanuelle Perret, Renée C. R. M. Vossen, Maria C. E. van Dam–Mieras, Donato Zipeto, Jean-Louis Virelizier, Phuc LeHoang, Cathrien A. Bruggeman, Susan Michelson; Entry of Human Cytomegalovirus into Retinal Pigment Epithelial and Endothelial Cells by Endocytosis. Invest. Ophthalmol. Vis. Sci. 1999;40(11):2598-2607.
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purpose. Human retinal pigment epithelial (RPE) cells and endothelial cells
(HUVECs) are targets of human cytomegalovirus (HCMV) infection in vivo
with significantly protracted replication in vitro compared with that
in fibroblasts. This study analyzes the kinetics and mechanisms of HCMV
entry into both cell types.
methods. RPE cells were obtained from donor eyes. HUVECs were isolated from
human umbilical cords. HCMV entrance was followed by electron
microscopy and immunofluorescence in the presence of lysosomotropic
agents and cytochalasin B.
results. Human cytomegalovirus entered into RPE cells and HUVECs as early as 5
minutes after virus–cell contact. Entry was mediated by endocytosis,
whereas HCMV enters fibroblasts through fusion. Most internalized viral
particles and dense bodies appeared to be degraded within vacuoles.
Viral entry, transport of viral proteins to the nucleus, and onset of
viral transcription (immediate early [IE] protein expression) were
significantly blocked by cytochalasin B. Lysosomotropic agents did not
significantly reduce IE expression in RPE cells or HUVECs.
conclusions. This study shows that HCMV penetrates these highly specialized relevant
cells via endocytosis. The low level of infection and the delay in the
onset of HCMV expression seen in these cells compared with fibroblasts
may be related to the sequestration and degradation of incoming viral
particles in endocytic vacuoles.
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