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Role for nectin‐1 in herpes simplex virus 1 entry and spread in human retinal pigment epithelial cells
Author(s) -
Tiwari Vaibhav,
Oh MyungJin,
Kovacs Maria,
Shukla Shripaad Y.,
ValyiNagy Tibor,
Shukla Deepak
Publication year - 2008
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2008.06655.x
Subject(s) - microbiology and biotechnology , biology , herpes simplex virus , filopodia , viral entry , retinal pigment epithelium , nectin , fascin , virology , virus , retina , cell , viral replication , cell adhesion , actin , genetics , neuroscience
Herpes simplex virus 1 (HSV‐1) demonstrates a unique ability to infect a variety of host cell types. Retinal pigment epithelial (RPE) cells form the outermost layer of the retina and provide a potential target for viral invasion and permanent vision impairment. Here we examine the initial cellular and molecular mechanisms that facilitate HSV‐1 invasion of human RPE cells. High‐resolution confocal microscopy demonstrated initial interaction of green fluorescent protein (GFP)‐tagged virions with filopodia‐like structures present on cell surfaces. Unidirectional movement of the virions on filopodia to the cell body was detected by live cell imaging of RPE cells, which demonstrated susceptibility to pH‐dependent HSV‐1 entry and replication. Use of RT‐PCR indicated expression of nectin‐1, herpes virus entry mediator (HVEM) and 3‐ O ‐sulfotransferase‐3 (as a surrogate marker for 3‐O‐sulfated heparan sulfate). HVEM and nectin‐1 expression was subsequently verified by flow cytometry. Nectin‐1 expression in murine retinal tissue was also demonstrated by immunohistochemistry. Antibodies against nectin‐1, but not HVEM, were able to block HSV‐1 infection. Similar blocking effects were seen with a small interfering RNA construct specifically directed against nectin‐1, which also blocked RPE cell fusion with HSV‐1 glycoprotein‐expressing Chinese hamster ovary (CHO‐K1) cells. Anti‐nectin‐1 antibodies and F‐actin depolymerizers were also successful in blocking the cytoskeletal changes that occur upon HSV‐1 entry into cells. Our findings shed new light on the cellular and molecular mechanisms that help the virus to enter the cells of the inner eye.