首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Molecular and biological characterization of a replication competent human immunodeficiency type 2 (HIV-2) proviral clone.
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Molecular and biological characterization of a replication competent human immunodeficiency type 2 (HIV-2) proviral clone.

机译:具有复制能力的人类2型免疫缺陷病毒(HIV-2)前病毒克隆的分子和生物学特征。

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摘要

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.
机译: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2型免疫缺陷病毒(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原病毒DNA的复制能力。一个基因组克隆,命名为HIV-2SBL / ISY,在培养转染细胞数周后产生逆转录病毒颗粒。 HIV-2SBL / ISY克隆包含完整的原病毒和细胞侧翼序列。我们获得了原病毒的DNA序列,并将其与其他两个HIV-2分离株的公开序列进行了比较。 HIV-2分离株之间的变异程度与迄今测序的非洲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)类似,但在亲本病毒中截短的包膜跨膜蛋白(gp32-34)不同,而在HIV-2SBL / ISY(gp41)中则不同。亲本病毒和克隆病毒都对某些人类T细胞系具有传染性和致细胞性,诱导合胞体并在体外感染人类巨噬细胞系(U937)。具有生物活性的HIV-2克隆的可用性为研究HIV-2基因在体外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克隆病毒体外感染恒河猴的新鲜外周血T细胞并体内感染同一动物,因此其在动物中的应用可能代表了体内病毒基因功能研究以及实验性开发的模型预防和治疗人类逆转录病毒感染的方法。

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