Human T-cell lymphotropic virus type 1 can infect primary rat retinal glial cells and induce gene expression of inflammatory cytokines

Abstract

PURPOSE. To examine whether or not retinal glial cells can be infected by human T-cell lymphotropic virus type 1 (HTLV-1) and test the possibility that HTLV-1-infected retinal glial cells are involved in the pathogenesis of HTLV-1 uveitis (HU). METHODS. We tested infection of HTLV-1 by a standard co-culturing method using WKAH rat retinal glial cells and irradiated MT-2, a human T cell line that produces HTLV-1. Infection was confirmed by detecting the integrated HTLV-1 provirus, using polymerase chain reaction (PCR), viral gene expression, using reverse transcriptase-PCR (RT-PCR) and HTLV-1 p19 ELISA, and by identifying the HTLV-1-infected glial cells by immunofluorescence cytochemistry and in situ hybridization. Changes in cytokine gene expression were studied by RT-PCR. RESULTS. Using a semiquantitative PCR of HTLV-1 provirus sequence, we found that 2.6% of the retinal glial cells were infected at 3 days after infection, followed by a gradual decrease in the percentage with an extended period of culture up to 4 weeks. This time course of infection was also verified by RT-PCR and ELISA studies that detect viral mRNA expression and protein production, respectively. Expression of HTLV-1 gag protein and tax mRNA was detected in a part of glial cells by indirect immunofluorescence cytochemistry and in situ hybridization, respectively. RT-PCR analysis of cytokine gene expression revealed that gene expression of IL-6, CINC-1 (Gro, KC), and TNF-a were induced in these cells, with a peak at 3 weeks after infection. CONCLUSION. These results provided supportive evidence for the theory that the infection of retinal glial cells by HTLV-1 and subsequent production of inflammatory cytokines could be one contributing factor for the development of the unique clinical features of HU. A better understanding of the specific roles of the inflammatory cytokines in the pathogenesis of HU would be beneficial in the treatment and control of this disease.