Further evaluation of soluble CD4 as an anti-HIV type 1 gene therapy: demonstration of protection of primary human peripheral blood lymphocytes from infection by HIV type 1.
We previously reported on the construction of retroviral vectors that produce a secreted form of the HIV-1 receptor, T cell antigen CD4 (Morgan et al., AIDS Res Hum Retroviruses 1990;6:183-191). In this article we test the ability of these sCD4-expressing retroviral vectors to protect human T-cell lines or primary T cells from HIV-1 infection. To demonstrate that protection from HIV-1 infection is mediated by the soluble nature of this protein, two coculture protection experiments were conducted. In these experiments, sCD4-expressing retroviral vectors were used to engineer mouse NIH 3T3 cells. In one coculture experiment the human SupT1 cell line was added directly to the culture of sCD4-producing NIH 3T3 cells, and in another experiment the two cell types were separated physically by a semipermeable membrane. In both coculture configurations, the T cell line was protected from HIV-1 challenge as measured by syncytium formation and indirect immunofluorescent assays. In addition, the SupT1 line was directly engineered with sCD4-expressing retroviral vectors and shown to be protected from HIV-1 challenge. As a prelude to further preclinical studies, we tested the ability of retroviral vectors to transduce primary human peripheral blood lymphocytes (PBLs). Conditions used to stimulate T cell growth resulted in significant shifts in the CD4/CD8 cell in favor of CD8 cells. Retroviral-mediated gene transfer under these conditions resulted in low levels of gene transfer (< 5%).(ABSTRACT TRUNCATED AT 250 WORDS)
Morgan
R A
RA
National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
Baler-Bitterlich
G
G
Ragheb
J A
JA
Wong-Staal
F
F
Gallo
R C
RC
Anderson
W F
WF
D016475:3T3 Cells; D000818:Animals; D015704:CD4 Antigens / Q000096:biosynthesis / Q000235:genetics* / Q000627:therapeutic use; D002460:Cell Line; D002478:Cells, Cultured; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015658:HIV Infections / Q000517:prevention & control* / Q000628:therapy; D015497:HIV-1; D006801:Humans; D008214:Lymphocytes / Q000821:virology; D051379:Mice; D012995:Solubility; D014161:Transduction, Genetic
D015704:CD4 Antigens