AEGiS-07IAC: Development of an HIV-1 infection system for studies on viral life cycle in neural cells and isolation of neurotropic HIV-1 strains.

7th International AIDS Conference

Florence, Italy — June 16-21, 1991

Development of an HIV-1 infection system for studies on viral life cycle in neural cells and isolation of neurotropic HIV-1 strains.

Int Conf AIDS 1991 Jun 16-21; 7:99 (abstract no. M.A.1030)
Volsky B, Sakai K, Shahabuddin M, Volsky DJ; Molecular Virology Laboratory, St.Luke's/Roosevelt Hospital Center, Columbia University, New York, N.Y.

OBJECTIVE: HIV-1 infects neural cells but major differences exist compared to its infection of T cells. The route of HIV-1 entry into neural cells is unknown, the infection is small in extent and non-cytopathic, and virus can persist in a dormant/inducible state. Our objective was to develop an in vitro neural cell system which is susceptible to efficient HIV-1 infection, and to employ it for studies on the HIV-1 life cycle in neural cells, its interruption with antiviral compounds, and for isolation of neurotropic HIV-1 strains.

METHODS: Neural cells were stably transfected with a CD4 expression vector pKS286. CD4+ and CD4- cells were infected with laboratory strains of HIV-1, and a natural isolate which could not be propagated in T cells. The infected cells were tested for: kinetics of expression of viral DNA, RNA, and proteins; relative levels of integrated and unintegrated viral DNA; cellular regulation of viral expression; sensitivity to antiviral compounds.

RESULTS: Stable transfection of glial HTB148 cells with the pKS286 vector generated a cell line, HTB148/286, which expressed high levels of surface CD4 receptors. Three days after infection with HIV-1, 20-30% of HTB148/286 cells expressed HIV-1 antigens and produced high titers of progeny virus. HIV-1 expression declined upon prolonged incubation. AZT was equally effective in blocking infection in these cells and T cells, suggesting that the early stages of HIV-1 infection were similar in both cell systems. In contrast, agents active in T cells in inhibition of transcription from DNA were significantly less effective in neural cells, reflecting the presence of distinct neural cell factors mediating HIV-1 transcription. The HTB148/286 cells supported the replication of a natural HIV-1 isolate which initially did not grow in T cells.

CONCLUSIONS: These data establish the utility of this model system for high expression of HIV-1 in neural cells. Cell-specific expression of some HIV-1 functions should be considered in the design of new therapeutics.

Keywords: AEGIS, HIV-1, Virus Replication, HIV Infections, Antigens, CD4, Zidovudine, DNA, Viral, Life Cycle Stages, CD4-Positive T-Lymphocytes, Cells, Anti-HIV Agents, In Vitro, growth & development, virology, therapy, surgery, drug therapy, ICA7
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