Franchini G, Fargnoli K A, Giombini F, Jagodzinski L, De Rossi A, Bosch M, Biberfeld G, Fenyo E M, Albert J, Gallo R C
Laboratory of Tumor Cell Biology, National Cancer Institute, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1989 Apr;86(7):2433-7. doi: 10.1073/pnas.86.7.2433.
We obtained complete genomic clones of human immunodeficiency virus type 2 (HIV-2) from the DNA of the neoplastic human cell line HUT 78 freshly infected with a HIV-2 isolate, strain SBL6669. The recombinant phage DNA was transfected into the lymphocytes of CD4-positive HUT 78 cell line to test the replication competence of the proviral DNA. One genomic clone, designated HIV-2SBL/ISY, yielded retroviral particles after a few weeks of culture of the transfected cells. The HIV-2SBL/ISY clone contained a complete provirus and cellular flanking sequence. We obtained the DNA sequence of the provirus and compared it with the published sequence of two other HIV-2 isolates. The degree of variability among HIV-2 isolates is comparable to that observed among African HIV-1 isolates sequenced to date. Immunologically, HIV-2SBL/ISY is similar to the parental virus (HIV-2SBL6669) but differs in the envelope transmembrane protein that is truncated (gp32-34) in the parental virus and not in HIV-2SBL/ISY (gp41). Both the parental and the cloned viruses are infectious and cytopathic for some human T-cell lines, induce syncytia, and infect a human macrophage cell line (U937) in vitro. The availability of a biologically active HIV-2 clone provides the means to study the role and interaction of HIV-2 genes in vitro as well as to assess the functional similarities among HIV-1 and HIV-2 genes. Since HIV-2SBL/ISY cloned virus infects fresh peripheral blood T cells from Rhesus macaques in vitro and infects the same animal in vivo, its use in animals may represent a model for functional study of viral genes in vivo as well as for development of experimental approaches to prevent and cure retroviral infection in humans.
我们从新感染了HIV-2分离株SBL6669的肿瘤性人类细胞系HUT 78的DNA中获得了人类免疫缺陷病毒2型（HIV-2）的完整基因组克隆。将重组噬菌体DNA转染到CD4阳性HUT 78细胞系的淋巴细胞中，以测试前病毒DNA的复制能力。一个基因组克隆，命名为HIV-2SBL/ISY，在转染细胞培养几周后产生了逆转录病毒颗粒。HIV-2SBL/ISY克隆包含一个完整的前病毒和细胞侧翼序列。我们获得了前病毒的DNA序列，并将其与另外两个HIV-2分离株的已发表序列进行了比较。HIV-2分离株之间的变异程度与迄今为止测序的非洲HIV-1分离株中观察到的变异程度相当。在免疫学上，HIV-2SBL/ISY与亲本病毒（HIV-2SBL6669）相似，但在包膜跨膜蛋白方面有所不同，亲本病毒中的该蛋白被截短（gp32 - 34），而HIV-2SBL/ISY中则没有（gp41）。亲本病毒和克隆病毒对一些人类T细胞系都具有感染性和细胞病变作用，能诱导形成多核巨细胞，并在体外感染人类巨噬细胞系（U937）。具有生物活性的HIV-2克隆的获得为在体外研究HIV-2基因的作用和相互作用以及评估HIV-1和HIV-2基因之间的功能相似性提供了手段。由于HIV-2SBL/ISY克隆病毒在体外能感染恒河猴的新鲜外周血T细胞，并在体内感染同一只动物，因此将其用于动物可能代表了一种在体内对病毒基因进行功能研究的模型，以及开发预防和治疗人类逆转录病毒感染的实验方法的模型。
