INHIBITION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REPLICATION BY NUCLEAR CHIMERIC ANTI-HIV RIBOZYMES IN A HUMAN T-LYMPHOBLASTOID CELL-LINE

Human immunodeficiency virus (HIV) infection represents one of the mos t challenging systems for gene therapy, Thanks to the extended knowled ge of the molecular biology of the HIV life cycle, many different stra tegies have been developed including transdominant modifications of HI V proteins, RNA decoys, antisense RNA, ribozymes, and intracellular an tibody fragments. In this paper, we have tested in a human T lymphobla stoid cell line the antiviral activity of ribozymes specifically desig ned to co-localize inside the nucleus with the Rev pre-mRNA before it is spliced and transported to the cytoplasm, This result was obtained by inserting the ribozyme in the spliceosomal U1 small nuclear RNA (sn RNA) and in a derivative that has perfect complementarity with the 5' splice site of the Rev pre-mRNA, These ribozymes were tested in human T cell clones and were shown to be very efficient in inhibiting viral replication, Not only were the p24 levels in the culture medium drasti cally reduced but so were the intracellular HIV transcripts, Control d isabled ribozymes enabled us to show the specificity of the ribozyme a ctivity, Therefore, these constructs have potential utility for gene t herapy of HIV-1 infection.