Federico M, Bona R, D'Aloja P, Baiocchi M, Pugliese K, Nappi F, Chelucci C, Mavilio F, Verani P
Laboratory of Virology, Istituto Superiore di Sanità, Rome, Italy.
Acta Haematol. 1996;95(3-4):199-203. doi: 10.1159/000203878.
A Hut-78 cell clone (F12) harboring a nonproducer human immunodeficiency virus (HIV-1) variant shows complete resistance to HIV-1 or HIV-2 superinfection. The F12-HIV provirus produces an altered HIV-1 protein pattern and cannot generate even immature viral particles. We demonstrated that HeLa CD4+ cells transfected with the F12-HIV genome resist HIV superinfection through a CD4-independent mechanism. As F12-HIV appears to be a useful system to induce anti-HIV intracellular immunization, we constructed various retroviral vectors containing the F12-HIV genome, modified by elimination of the F12 3'LTR and part of its nef gene, inserted 'antisense' with respect to the Moloney murine leukemia virus 5' LTR. Here we show that recombinant retroviral particles carrying the N2/F12-HIV nef- (as) construct can stably transduce both CEMss human cells and primary human peripheral blood lymphocytes, inducing the expression of the F12-HIV genome. These results could open the way to an anti-AIDS gene therapy strategy based on F12-HIV-induced intracellular immunization.
一个携带无病毒产生能力的人类免疫缺陷病毒（HIV-1）变异体的Hut-78细胞克隆（F12）对HIV-1或HIV-2的超感染表现出完全抗性。F12-HIV前病毒产生了改变的HIV-1蛋白模式，甚至无法产生未成熟的病毒颗粒。我们证明，用F12-HIV基因组转染的HeLa CD4+细胞通过一种不依赖CD4的机制抵抗HIV超感染。由于F12-HIV似乎是诱导抗HIV细胞内免疫的有用系统，我们构建了各种逆转录病毒载体，这些载体包含通过去除F12 3'LTR及其部分nef基因进行修饰的F12-HIV基因组，并相对于莫洛尼鼠白血病病毒5'LTR反向插入。在此我们表明，携带N2/F12-HIV nef-（反义）构建体的重组逆转录病毒颗粒能够稳定转导CEMss人细胞和原代人外周血淋巴细胞，诱导F12-HIV基因组的表达。这些结果可能为基于F12-HIV诱导的细胞内免疫的抗艾滋病基因治疗策略开辟道路。
