INHIBITION OF HIV-1 MULTIPLICATION IN A HUMAN CD4-LINE EXPRESSING ANTISENSE AND SENSE RNA MOLECULES CONTAINING HIV-1 PACKAGING SIGNAL AND REV RESPONSE ELEMENT(S)( LYMPHOCYTIC CELL)

Moloney murine leukemia virua (MoMuLV)-derived retroviral vectors were engineered to express human immunodeficiency virus type 1 (HIV-1) pac kaging (psi) signal and Rev response element (RRE) sequences in either sense or antisense orientation. The RRE sequences were expressed unde r the control of the herpes simplex virus (HSV) thymidine kinase (tk) promoter fused to the HIV-1 trans-activation-responsive (TAR) element, while the psi signal sequences were expressed under control of the HS V tk promoter. Both RRE and psi signal sequences were expressed as par t of the 3' untranslated region of the neomycin phosphotransferase (ne o) mRNA. The constructs were used to transfect/infect packaging cell l ines and the retroviral vector particles released were used to infect a human CD4+ lymphocyte-derived MT4 cell line. The stable MT4 transfor mants, harboring proviral vector DNA expressing one to two copies of H IV-1 RRE and psi signal in either antisense or sense orientation, were each tested for their susceptibility to HIV-1 infection. Compared to the results obtained with the control cells lacking any of the test DN A sequences, the rate of HIV-1 production remained unaltered in RRE1(sense RNA containing a single copy of RRE) RNA-containing cells, wher eas ft was delayed in cells expressing both RRE2+ (sense RNA containin g two copies of RRE) and RRE1- (antisense RNA containing a single copy of RRE) RNA-expressing cells. In cells expressing HIV-1 psi signal, H IV-1 production remained unaltered in psi+ RNA-expressing cells, where as it was delayed by up to 30 days in psi- RNA-expressing cells. These results indicate that retroviral vectors expressing two copies of HIV -1 RRE in sense orientation, a single copy of HIV-1 RRE in antisense o rientation, and particularly the HIV-1 psi signal in antisense orienta tion can be used to confer HIV-1 resistance.