TAT AND NEF DEFECTIVE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REPLICATES EFFICIENTLY IN TAT-EXPRESSING T-CELLS

Abstract

Human immunodeficiency virus type 1 (HIV-1) contains several nonstructural genes which are required for viral replication and disease pathogenesis. Among them, rat and nef genes encode an essential transactivator of HIV-1 LTR and a pluripotent protein which seems to be essential for in vivo but not in vitro viral replication, respectively. We constructed two tat and nef defective HIV-1 and tested for their abilities Co replicate in several T cells. The defective viruses did not replicate in CD4(+) T cells, but were rescued in the recombinant Jurkat-tat cells which express the rat gene. The replication of HXB/tnC which is defective in far and nef, and contains the chloramphenicol acetyltransferase (CAT) gene was easily detected by a CAT assay. No revertant was identified during the passages of the mutant-viruses for more than two months in Jurkat-tat cells, far and nef defective HIV-1 could be used instead of wild type viruses for several purposes such as inhibitor screening and development of killed AIDS vaccine.