ENHANCEMENT OR INHIBITION OF HIV-1 REPLICATION BY INTRACELLULAR EXPRESSION OF SENSE OR ANTISENSE RNA TARGETED AT DIFFERENT INTERMEDIATES OFREVERSE TRANSCRIPTION

Objectives: To construct retroviral vectors expressing sense or antise nse RNA targeted at HIV reverse transcription intermediates, and to te st the anti-HIV properties of these constructs in transduced T cells. Design: Five double-copy retroviral vectors were constructed, in which the expression of the sense or antisense RNA corresponding to HIV min us- or plus-strand strong-stop DNA was driven by the human tRNA(met) p romoter. Method: The templates for the sense or antisense RNA were pol ymerase chain reaction-cloned from HIV pNL43 into a murine leukaemia v irus-based vector and corresponding defective virions were packaged in PA317 cells. Human jurkat T cells transduced with these vectors were challenged with HIV and monitored for viral RNA, viral DNA and p24 pro duction for 23 weeks. Results: Intracellular expression of HIV sense R U5 sequences (RNA complementary to minus-strand strong-stop DNA) enhan ced HIV replication in T cells. Expression of HIV sense or antisense U 3RU5 sequences (identical or complementary to plus-strand strong-stop DNA) conferred long-term inhibition of HIV replication, despite contin uous presence of viral challenge in the transduced cell cultures. Conc lusion: Plus-strand strong-stop DNA as an intermediate in the early pr ocess of viral reverse transcription can be explored as an additional target for anti-HIV gene therapy.