In Vitro Selection of HIV-1 Resistant to an Anti-CD4 Monoclonal Antibody That Inhibits Virus Transcription

Abstract

Phase I studies using monoclonal antibodies (mAbs) that bind to the Ig-CDR3-like loop in domain 1 of CD4 (e.g., 13B8-2 mAb) have already been documented for HIV-1-infected patients. In vitro, such mAbs do not inhibit virus to cell fusion but are able to inhibit virus envelope-mediated syncytia formation. Moreover, these mAbs inhibit Tat-induced activation of HIV-1 promoter and HIV-1 transcription in infected CD4+ cells. Here, we report the selection of escaped mutant virus or viruses derived from HIV-1Lai capable of replicating in vitro in the presence of concentrations of 13B8-2 mAb, that usually inhibit HIV-1Lai particle production. The escaped mutant virus or viruses, termed HIV-1Lai13EM, kept the major enzymatic restriction sites found in HIV-lLai and remained sensitive to anti-CD4 mAb-, soluble CD4-, and recombinant gpl20-mediated inhibition of syncytia formation. Possible genetic changes affecting the tat gene or the 5′ long terminal repeat (LTR) were investigated. Partial sequence analysis of HIV-1Lai13EM and a control HIV-lLai grown for 85 days in CEM cells, demonstrated that the first tat exon of these two viruses encoded identical proteins. Although a point mutation G>A was frequently encountered (6 of 13 sequences) in the LTRs of HIV-1Lai13EM at position -188 within the negative regulatory element (NRE), this mutation did not confer the escape mutant phenotype. Our study indicates that the mutant phenotype probably requires genetic changes in a region or regions outside the LTRs.