Structure and Origin of HIV Type 1 DNA in Persistently Infected B Lymphoblastoid Cell Lines

Abstract

The Epstein-Barr virus (EBV) and human immunodeficiency virus type 1 (HIV-1) can coinfect resting B cells, leading to EBV-carrying lymphoblastoid cell lines (LCLs) persistently infected with HIV-1. LCLs established from coinfected peripheral blood lymphocytes (PBLs) differed from LCLs derived from HIV-1-infected cell lines, in that the majority if not all of the cells expressed gp 120 and a high percentage produced infectious HIV-1 after continuous passage for 6-9 months. Restriction analysis of the putative HIV-1 provirus revealed that persistently infected LCLs carried variable copies of primarily unintegrated circular and/or linear forms of HIV-1 DNA. This extrachromosomal location is strikingly different from that of the one to three copies of integrated proviral DNA deleted in persistently infected T cell and monocytic cell lines. Anti-gp120 monoclonal antibody and 3′-azido-3′-deoxythymidine (AZT) inhibited HIV-1 expression and reduced HIV-1 DNA copy number in persistently infected LCLs, supporting the hypothesis that unintegrated HIV-1 DNA accumulates primarily as a result of superinfection. We propose that the extrachromosomal location of the HIV-1 DNA contributes to the semipermissive nature of B cell infection by HIV-1.