MOLECULAR CLONING AND CHARACTERIZATION OF RETROVIRUSES ASSOCIATED WITH AIDS

本项目的目标是获得有关 艾滋病毒在其生物化学、结构和多样性方面， 致病性和免疫变异性。 异质性 逆转录病毒基因组是慢病毒的独特特征。 人类 免疫缺陷病毒1型慢病毒（HIV-1），病原体 获得性免疫缺陷综合征，已被分离出来， CD 4阳性外周血T淋巴细胞（PBL）和单核细胞源性 巨噬细胞（MDM）的血清阳性个体。 稳定感染 HIV-1的分子克隆，它产生高滴度的病毒， 感染培养的PBL、MDM和多种CD 4阳性T细胞 白血病细胞系已被广泛用于描绘结构， HIV-1编码蛋白的功能。 然而，只有少数分子 克隆已经直接从感染的组织中分离出来。 这将是 有利的是获得这样的克隆而不必在组织中传代 文化和减少的概率拿起突变或 选择适应组织培养条件的变体。 我们 能够直接从SIV-HIV（SHIV）获得的肺组织中克隆 感染的、CD 4耗尽的食蟹猴，并获得6个个体 克隆 一种高度致细胞病变和形成合胞体的HIV-1病毒分离自 乌干达一名患者的血液， 感染人类PBL。 分离到9个完整的感染性克隆 从这个单一的隔离。 有趣的是， 克隆的病毒能够在人类PBL、黑猩猩PBL、MT-4 细胞（连续T细胞系）、H-9细胞和MDM。 这种分子 分离物是双嗜性，就其感染不同 细胞系

The objective of this project is to obtain information pertaining to the structure and diversity of HIV with respect to its biochemical, pathogenic, and immunological variability. Heterogeneity among retroviral genomes is a distinct feature of Lentiviruses. The human immunodeficiency virus type 1 Lentivirus (HIV-1), the causative agent of acquired immunodeficiency syndrome, has been isolated from CD4-positive peripheral blood T lymphocytes (PBLs) and monocyte-derived macrophages (MDMs) of seropositive individuals. Stable infectious molecular clones of HIV-1, which generate high titers of virus capable of infecting cultured PBLs, MDMs and a variety of CD4 positive T cell leukemic lines have been extensively used to delineate the structure and function of HIV-1 encoded proteins. However, only a few molecular clones have been isolated directly from infected tissue. It would be advantageous to obtain such clones without having to passage in tissue culture and lessening the probability of picking up mutations or selecting for variants that adapt to tissue culture conditions. We were able to clone directly from lung tissue obtained from a SIV-HIV (SHIV) infected, CD4 depleted cynomolgus macaque and obtained six individual clones. A highly cytopathic and syncytia forming HIV-1 virus was isolated from the blood of a patient from Uganda and was molecularly cloned from infected human PBLs. Nine complete and infectious clones were isolated from this single isolate. Of interest is the fact that the molecularly cloned viruses were able to grow in human PBLs, chimpanzee PBLs, MT-4 cells (continuous T-cell line), H-9 cells, and MDMs. This molecular isolate is dual tropic with respect to its ability to infect different cell lines.