Trans-activated interferon-alpha2 delivered to T cells by SV40 inhibits early stages in the HIV-1 replicative cycle.
Several lines of evidence suggest a potential major role for interferon (IFN) in controlling HIV-1 replication. However, this inhibition is moderate and is reversible upon IFN removal. To achieve prolonged high concentrations of IFN at the site of infection, we devised an SV40-based vector, SV[HIVLTR]IFN, to direct the synthesis of human IFN-alpha2, by employing a virus-trans-activated human IFN-alpha2 gene to be transcribed in response to HIV-1 infection. Expression of IFN-alpha2 was confirmed by Northern and Western blotting, in SV[HIVLTR]IFN-transduced, HIV-1-challenged human lymphocyte lines and primary human lymphocytes. SV[HIVLTR]IFN-transduced cells showed no evidence of HIV-1-related cytophatic effects when challenged with high doses of HIV-1(NL4-3). As measured by supernatant HIV-1 p24 antigen concentration, IFN-alpha2-expressing cell lines and peripheral blood lymphocytes (PBL) were protected from high-dose challenges of HIV-1. rSV40-delivered IFN-alpha2 inhibited gp120 protein synthesis and expression of HIV-1 mRNAs. Finally, Southern analysis revealed that levels of proviral DNA were markedly reduced in SV[HIVLTR]IFN-transduced cells compared to control cultures. IFN-alpha2 expression driven by HIVLTR delivered by an rSV40 vector thus strongly inhibits HIV-1 replication, probably by blocking a preintegration step in HIV-1 infection. Targeted expression of IFN-alpha2 delivered by SV40 can thus repress HIV-1 replication, and may be a useful approach to HIV-1 treatment.
Cordelier
Pierre
P
Department of Pathology, Jefferson Medical College, Philadelphia, PA 19107, USA.
Calarota
Sandra A
SA
Strayer
David S
DS
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D019556:COS Cells; D002460:Cell Line; D002478:Cells, Cultured; D004279:DNA, Viral / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D016655:HIV Core Protein p24 / Q000276:immunology; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis; D015497:HIV-1 / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use*; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011533:Proviruses / Q000502:physiology; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013539:Simian virus 40 / Q000378:metabolism / Q000502:physiology*; D013601:T-Lymphocytes / Q000276:immunology*; D013997:Time Factors; D015533:Transcriptional Activation
D004279:DNA, Viral; D016655:HIV Core Protein p24; D015699:HIV Envelope Protein gp120; D016898:Interferon-alpha; D012333:RNA, Messenger; D012313:RNA