Devare S G, Srinivasan A, Bohan C A, Spira T J, Curran J W, Kalyanaraman V S
Proc Natl Acad Sci U S A. 1986 Aug;83(15):5718-22. doi: 10.1073/pnas.83.15.5718.
We have isolated retroviruses from six acquired immunodeficiency syndrome (AIDS) and three lymphadenopathy syndrome (LAS) patients by cocultivation of patients' lymphocytes with phytohemagglutinin-stimulated normal T cells. In an effort to address the extent to which these viruses have identical genetic information or there is divergence in their genomic sequences, we have compared the nine retrovirus isolates by the following criteria: (i) antigenic cross-reactivity by highly specific and sensitive competition radioimmunoassay for the major internal antigen; (ii) restriction site mapping analysis; and (iii) immunoblot analysis and metabolic labeling of viral structural proteins and their analysis by polyacrylamide gel electrophoresis. The data indicate that individual retroviruses have significant restriction site polymorphism in their genome even though they were isolated from patients residing at one geographic location. Furthermore, this polymorphism is reflected in the variation of the apparent size of the gag and env gene-encoded structural proteins. The heterogeneity in AIDS retrovirus-encoded proteins may be due to either substitutions in the primary amino acid sequence of the protein or deletions or additions in the coding regions of proteins. The alterations in viral structural proteins among various AIDS retroviruses could have important implications in antigenic properties and/or pathogenicity in development of the disease, its detection, and ultimately its eradication.
我们通过将患者的淋巴细胞与植物血凝素刺激的正常T细胞共培养,从6名获得性免疫缺陷综合征(AIDS)患者和3名淋巴结病综合征(LAS)患者中分离出了逆转录病毒。为了探究这些病毒在多大程度上具有相同的遗传信息,或者它们的基因组序列是否存在差异,我们按照以下标准对这9株逆转录病毒分离株进行了比较:(i)通过针对主要内部抗原的高度特异性和灵敏的竞争放射免疫测定法进行抗原交叉反应性检测;(ii)限制性酶切位点图谱分析;(iii)免疫印迹分析以及病毒结构蛋白的代谢标记,并通过聚丙烯酰胺凝胶电泳对其进行分析。数据表明,尽管这些逆转录病毒是从居住在同一地理位置的患者中分离出来的,但单个逆转录病毒在其基因组中仍具有显著的限制性酶切位点多态性。此外,这种多态性反映在gag和env基因编码的结构蛋白的表观大小变化上。AIDS逆转录病毒编码蛋白的异质性可能是由于蛋白质一级氨基酸序列的替换,或者蛋白质编码区域的缺失或添加所致。不同AIDS逆转录病毒之间病毒结构蛋白的改变可能对疾病发展过程中的抗原特性和/或致病性、疾病的检测以及最终的根除具有重要意义。