Comparative analyses of intracellularly expressed antisense RNAs as inhibitors of human immunodeficiency virus type 1 replication.
The antiviral activities of intracellularly expressed antisense RNAs complementary to the human immunodeficiency virus type 1 (HIV-1) pol, vif, 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 promoter-directed retroviral transcript were constructed. The CD4+ T-cell line CEM-SS was transduced with retroviral constructs, and Northern blot 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, leading to 2- to 3-log10 reductions in p24 antigen production even at high inoculation doses (4 x 10(4) 50% tissue culture infective doses) of the HIV-1 strain HXB3. The strong antiviral effect correlated with a reduction of HIV-1 steady-state RNA levels, and with intracellular Tat protein production, suggesting that antisense transcripts act at an early step of HIV-1 replication. A lower steady-state antisense RNA level was detected in transduced primary CD4+ lymphocytes than in CEM-SS cells. Nevertheless, replication of the HIV-1 JR-CSF isolate was reduced with both the pol and env antisense RNA. Intracellularly expressed antisense sequences demonstrated more pronounced antiviral efficacy than the transdominant RevM10 protein, making these antisense RNAs a promising gene therapy strategy for HIV-1.
Veres
G
G
Systemix Inc., a Novartis Company, Palo Alto, California 94304, USA. gveres@stem.com
Junker
U
U
Baker
J
J
Barske
C
C
Kalfoglou
C
C
Ilves
H
H
Escaich
S
S
Kaneshima
H
H
Böhnlein
E
E
D016475:3T3 Cells; D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D002460:Cell Line; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015752:Genes, env; D015751:Genes, pol; D015757:Genes, rev; D016341:Genes, vif; D005822:Genetic Vectors; D016655:HIV Core Protein p24 / Q000032:analysis; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D051379:Mice; D008979:Moloney murine leukemia virus; D009690:Nucleic Acid Conformation; D016372:RNA, Antisense / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014779:Virus Replication / Q000187:drug effects*
D019380:Anti-HIV Agents; D016655:HIV Core Protein p24; D016372:RNA, Antisense