6th International AIDS Conference


San Francisco, California, USA — June 20-23, 1990

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HIV-1-infectivity bioassay.

Int Conf AIDS 1990 Jun 20-23; 6:341 (abstract no. 1109)
Pichler CE, Bettens F, Pichler WJ, Felber BK, Schwartz S, Pavlakis GN, de Weck AL; Institute for Clinical Immunology, Inselspital, Bern, Switzerland

OBJECTIVES: Short time bioassay to monitor HIV-1 virus infectivity in low numbers (less than 10(6)) of PBMC.

METHODS: 10(6) PBMC of HIV-1 infected individuals were stimulated with the anti-CD3 MoAb BMAO30 F(ab)2 fragment crosslinked to the anti-CD4 MoAb (Clonab T4),(=dual antibody stimulation). Thereby selective stimulation of only CD4+ T cells is achieved. HIV-1 replication was measured with a p24 enzyme immunoassay. To test HIV-1 infectivity, stimulated PBMC were cocultivated 2 days with the monocytic target cell line BF-24 (Science (1988), 239:184). The BF-24 cells have stably integrated copies of HIV-1 LTR promotor linked to the CAT gene (Chloramphenicol-acetyl-transferase). After HIV-1 infection, the virally encoded Tat proteins induce transcription of the CAT enzyme gene.

RESULTS: Selective stimulation of CD4+ cells in PBMC cultures of HIV-1 individuals enables to detect replicating virus 3 to 4 days after stimulation. In comparison, simultaneous stimulation of CD4+ and CD8+ T cells of HIV-1 individuals required longer in vitro cultures or inhibited detection of HIV-1 replication (p24 assay) and of HIV-1 infectivity (CAT enzyme assay).

CONCLUSION: Direct and selective stimulation of PBMC from HIV-1 infected individuals is possible and allows measurement of HIV-1 replication and infectivity without addition of allogene activated PBMC.

Keywords: AEGIS, HIV-1, HIV Infections, Virus Replication, HIV Long Terminal Repeat, Antigens, CD4, Gene Products, tat, Antigens, CD8, T-Lymphocytes, Monocytes, Cats, Animal, In Vitro, virology, genetics, ICA6KWDaegis,hiv-1,hivinfections,virusreplication,hivlongterminalrepeat,antigens,cd4,geneproducts,tat,antigens,cd8,t-lymphocytes,monocytes,cats,animal,invitro,virology,genetics,ica6

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