tailieunhanh - Báo cáo khoa học: Role for nectin-1 in herpes simplex virus 1 entry and spread in human retinal pigment epithelial cells

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 inva-sion and permanent vision impairment. | ễFEBS Journal Role for nectin-1 in herpes simplex virus 1 entry and spread in human retinal pigment epithelial cells Vaibhav Tiwari1 Myung-Jin Oh1 Maria Kovacs1 Shripaad Y. Shukla1 Tibor Valyi-Nagy2 and Deepak Shukla1 3 1 Department of Ophthalmology and VisualSciences College of Medicine University of Illinois Chicago IL USA 2 Department of Pathology College of Medicine University of Illinois Chicago IL USA 3 Department of Microbiology and Immunology College of Medicine University of Illinois Chicago IL USA Keywords actin cytoskeleton filopodia herpes simplex virus 1 human retinalpigment epithelial cells nectin-1 Correspondence D. Shukla Department of Ophthalmology VisualSciences 1855 West Taylor Street M C648 Chicago IL 60612 USA Fax 1 312 996 7773 Tel 1 312 355 0908 E-mail dshukla@ Received 16 June 2008 revised 5 August 2008 accepted 22 August 2008 doi 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 .