Transfer of an anti-HIV-1 ribozyme gene into primary human lymphocytes.
We reported previously that human CD4+ T cell lines stably expressing a hairpin ribozyme targeted to the human immunodeficiency virus type 1 (HIV-1) U5 leader sequence were resistant to challenge with diverse HIV-1 viral clones and clinical isolates (Yamada et al., 1994). To simulate more closely the in vivo infection process for investigations of anti-HIV-1 ribozyme gene therapy, we developed a system to transfer this ribozyme gene into freshly isolated human peripheral blood lymphocytes (PBLs) using a murine retrovirus vector. Following transduction and G418 selection, human PBLs from multiple donors expressed the ribozyme and resisted challenge by HIV-1 viral clones and clinical isolates, while control vector-transduced PBLs remained fully permissive for HIV-1 infection. No inhibition of an HIV-2 clone lacking the target was seen in ribozyme-expressing PBLs. Ribozyme expression had no effect on viability or proliferation kinetics of the primary lymphocytes. This study is the first demonstration in primary human T cells of resistance to HIV-1 infection conferred by gene transfer. A human clinical trial is in development to test further the safety and efficacy of this ribozyme in PBLs of HIV-1-infected patients in vivo.
Department of Medicine, University of California, San Diego, La Jolla 92093-0665.
M C
MC
Leavitt
M
M
Yu
O
O
Yamada
G
G
Kraus
D
D
Looney
E
E
Poeschla
F
F
Wong-Staal
D001483:Base Sequence; D002478:Cells, Cultured; D004351:Drug Resistance / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D015498:HIV-2 / Q000502:physiology; D006801:Humans; D007113:Immunity, Innate; D008214:Lymphocytes / Q000821:virology; D008969:Molecular Sequence Data; D012320:RNA Polymerase III / Q000378:metabolism; D016337:RNA, Catalytic / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012641:Selection, Genetic; D014779:Virus Replication
D005839:Gentamicins; D016337:RNA, Catalytic; D011993:Recombinant Fusion Proteins; C010680:antibiotic G 418; D012320:RNA Polymerase III