Jin Qingwen, Marsh Jon, Cornetta Kenneth, Alkhatib Ghalib
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Indiana University Vector Production Facility, Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
J Gen Virol. 2008 Oct;89(Pt 10):2611-2621. doi: 10.1099/vir.0.2008/003624-0.
It has previously been demonstrated that there are two distinct mechanisms for genetic resistance to human immunodeficiency virus type 1 (HIV-1) conferred by the CCR5Delta32 gene: the loss of wild-type CCR5 surface expression and the generation of CCR5Delta32 protein, which interacts with CXCR4. To analyse the protective effects of long-term expression of the CCR5Delta32 protein, recombinant lentiviral vectors were used to deliver the CCR5Delta32 gene into human cell lines and primary peripheral blood mononuclear cells that had been immortalized by human T-cell leukemia virus type 1. Blasticidin S-resistant cell lines expressing the lentivirus-encoded CCR5Delta32 showed a significant reduction in HIV-1 Env-mediated fusion assays. It was shown that CD4(+) T lymphocytes expressing the lentivirus-encoded CCR5Delta32 gene were highly resistant to infection by a primary but not by a laboratory-adapted X4 strain, suggesting different infectivity requirements. In contrast to previous studies that analysed the CCR5Delta32 protective effects in a transient expression system, this study showed that long-term expression of CCR5Delta32 conferred resistance to HIV-1 despite cell-surface expression of the HIV co-receptors. The results suggest an additional unknown mechanism for generating the CCR5Delta32 resistance phenotype and support the hypothesis that the CCR5Delta32 protein acts as an HIV-suppressive factor by altering the stoichiometry of the molecules involved in HIV-1 entry. The lentiviral-CCR5Delta32 vectors offer a method of generating HIV-resistant cells by delivery of the CCR5Delta32 gene that may be useful for stem cell- or T-cell-based gene therapy for HIV-1 infection.
先前已经证明,CCR5Δ32基因赋予对1型人类免疫缺陷病毒(HIV-1)遗传抗性的机制有两种:野生型CCR5表面表达的丧失以及与CXCR4相互作用的CCR5Δ32蛋白的产生。为了分析CCR5Δ32蛋白长期表达的保护作用,使用重组慢病毒载体将CCR5Δ32基因导入人类细胞系和由1型人类T细胞白血病病毒永生化的原代外周血单核细胞。表达慢病毒编码的CCR5Δ32的杀稻瘟菌素S抗性细胞系在HIV-1 Env介导的融合试验中显示出显著降低。结果表明,表达慢病毒编码的CCR5Δ32基因的CD4(+) T淋巴细胞对原发性但非实验室适应的X4毒株的感染具有高度抗性,提示不同的感染性要求。与先前在瞬时表达系统中分析CCR5Δ32保护作用的研究不同,本研究表明,尽管HIV共受体在细胞表面表达,但CCR5Δ32的长期表达仍赋予对HIV-1的抗性。结果提示产生CCR5Δ32抗性表型的另一种未知机制,并支持CCR5Δ32蛋白通过改变参与HIV-1进入的分子的化学计量比作为HIV抑制因子的假说。慢病毒-CCR5Δ32载体提供了一种通过递送CCR5Δ32基因产生HIV抗性细胞的方法,这可能对基于干细胞或T细胞的HIV-1感染基因治疗有用。