Abstract
Abstract: :
Purpose: To study the regulation of transcription of Chemokine receptor genes in corneal and retinal pigment epithelial cell lines during infection with Herpes Simplex virus-1 (HSV-1). Methods: Human corneal epithelial cell line (HCET) and retinal epithelial cell line (ARPE) were infected with a laboratory-adapted strain (F1) and a clinical isolate of HSV-1. RNA from HSV infected and uninfected cells were purified and the transcripts of Chemokine receptors and a house keeping gene, GADPH were analyzed by multiplex RT-PCR to detect six targets in a single tube reaction at the same time. The DNA from PCR was resolved by agarose gel electrophoresis to find the qualitative differences in the expression of the chemokine receptors in the infected and uninfected cells. Results: HSV-1 infection results in up-regulation of transcripts of the chemokine receptors CXCR1, CXCR2, CXCR4, CCR4, CCR6, CCR7, CCR8 and V28 in both HCET and ARPE cells. In addition, the transcripts of CXCR3 and CCR5 (which are expressed at either low levels or not expressed in HCET cells respectively), are up-regulated in HSV-1 infected ARPE cells. CCR9 is expressed in both uninfected and infected HCET cells, and in uninfected ARPE cells, while it is down regulated in HSV-1 infected ARPE cells. . Later in infection CXCR4, CCR8 and V28 are down regulated only in ARPE cells, but not in HCET cells suggesting that up-regulation is mediated by HSV-immediate early / early genes. The mRNA coding for SDF, the ligand for the chemokine receptor, CXCR 4, is not expressed in HCET cells and uninfected ARPE cells while it is up-regulated in HSV infected ARPE cells. ELISA data reveals that HSV-1 infected cells do not produce MDC or TARC. Conclusion: We have shown differential cell specific up-regulation of several chemokine receptors in HSV-1 infected ocular cells.. These data suggest that chemokine ligands expressed by infiltrating cells have the potential to alter intracellular signaling upon binding to the chemokine receptors that are up-regulated during ocular HSV-1 infections.
Keywords: 425 herpes simplex virus • 372 cornea: epithelium • 567 retinal pigment epithelium