COMPARATIVE ANALYSES OF INTRACELLULARLY EXPRESSED ANTISENSE RNAS AS INHIBITORS OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REPLICATION

The antiviral activities of intracellularly expressed antisense RNAs c omplementary to the human immunodeficiency virus tape 1 (HIV-1) pol, v if, and env genes and the 3' long terminal repeat (LTR) sequence were evaluated in this comparative study, Retroviral vectors expressing the antisense RNAs as part of the Moloney murine leukemia virus LTR promo ter-directed retroviral transcript were constructed. The CD4(+) T-cell line CEM-SS was transduced with retroviral constructs, and Northern b lot analyses showed high steady-state antisense RNA expression levels, The most efficient inhibition of HIV-1 replication was observed with the env antisense RNA, followed by the pol complementary sequence, lea ding to 2- to 3-log(10) reductions in p24 antigen production even at h igh inoculation doses (4 x 10(4) 50% tissue culture infective doses) o f the HIV-1 strain HXB3. The strong antiviral effect correlated with a reduction of HIV-1 steady-state RNA levels, and with intracellular Ta t protein production, suggesting that antisense transcripts act at an early step of HIV-1 replication. A lower steady-state antisense RNA le vel was detected in transduced primary CD4(+) lymphocytes than in CEM- SS cells, Nevertheless, replication of the HIV-1 JR-CSF isolate was re duced with both the pol and env antisense RNA, Intracellularly express ed antisense sequences demonstrated more pronounced antiviral efficacy than the trans-dominant RevM10 protein, making these antisense RNAs a promising gene therapy strategy for HIV-1.