ISOLATION AND LONG-TERM CULTURE OF PRIMARY OCULAR HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 ISOLATES IN PRIMARY ASTROCYTES

Vitreous specimens from 14 HIV-1 infected persons undergoing medically indicated vitrectomy were assayed for the presence of infectious HIV- 1 and viral tropism. Human primary fetal astrocytes, adult lymphocytes , or macrophages were exposed to vitreous in culture and cells were th en assayed for HIV-1 DNA by polymerase chain reaction amplification. W e found that 11 of 14 patients tested carried ocular HIV-1 which repli cated in one or more primary cell types; of the 13 vitreous samples te sted in astrocytes, eight contained transmissible HIV-1. The three pat ients with no culturable ocular virus were in antiviral therapy at the time of vitrectomy. Comparison of envelope V3 sequences from astrocyt es infected in culture to that in uncultured blood cells revealed 21% sequence divergence indicating that ocular HIV-1 transmitted to astroc ytes was not recently derived from virus present in the blood. Two ocu lar samples transmissible to astrocytes were tested further and found capable of sustained replication by serial passage to uninfected astro cytes. However, the viral structural proteins produced by infected ast rocytes were abnormal, p24 was absent and higher molecular weight Gag proteins were present. We conclude that the eye is a central nervous s ystem compartment which frequently contains HIV-1 cap able of replicat ion in human astrocytes.