A system for the high efficiency replication of HIV-1 in neural cells and its application to anti-viral evaluation.
Stable transfection of H4 neuroglioma cells with the Epstein-Barr virus-based eucaryotic CD4 expression vector pKS286 generated the cell line, H4/CD4, in which greater than 90% of cells express surface CD4 receptors. Optimal conditions for infection of H4/CD4 cells with HIV-1 were determined; these included a cocultivation with growth-arrested, chronically infected T cells. Under these conditions, 3-days after infection up to 50% of H4/CD4 cells expressed HIV-1 antigens as detected by immunofluorescence assay, the number of intracellular HIV-1 RNA copies reached 10(3) molecules per cell as determined by liquid hybridization, and virus production ranged from 0.2 to 1.0 micrograms HIV-1 p24 core antigen per ml of culture supernatant, comparable to that measured under the same conditions in HIV-1 infected T cells. Giant cells and cytolysis were common. Inhibition of HIV-1 infection by nucleoside analogues in H4/CD4 cells was comparable to that in T cells, suggesting that the early stages of HIV-1 infection were similar in both cell systems. Infection in the presence of soluble CD4 reduced HIV-1 expression to the levels determined in CD4-negative H4 cells. This system may be useful for screening of drugs intended to block HIV-1 replication in the brain and for the evaluation of the HIV-1 life cycle in brain cells.
Volsky
B
B
Division of Molecular Virology, St Luke's/Roosevelt Hospital Center, Columbia University, New York, New York 10019.
Sakai
K
K
Reddy
M M
MM
Volsky
D J
DJ
D015704:CD4 Antigens / Q000235:genetics; D002459:Cell Fusion; D002478:Cells, Cultured; D016655:HIV Core Protein p24 / Q000032:analysis; D015658:HIV Infections / Q000188:drug therapy; D015497:HIV-1 / Q000254:growth & development*; D006801:Humans; D066298:In Vitro Techniques; D009474:Neurons / Q000382:microbiology*; D012367:RNA, Viral / Q000032:analysis; D011994:Recombinant Proteins; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects; D016047:Zalcitabine / Q000494:pharmacology; D015215:Zidovudine / Q000494:pharmacology
D015704:CD4 Antigens; D016655:HIV Core Protein p24; D012367:RNA, Viral; D011994:Recombinant Proteins; D015215:Zidovudine; D016047:Zalcitabine