EVOLUTION OF AN ACUTELY LETHAL SIV

描述：（改编自申请人的摘要）为了获得更好的 了解人类免疫缺陷病毒（HIV）的发病机制， 研究者建议使用猕猴的感染， 分离自白眉猴的相关猿猴病毒（SIVsmm）。 博士 Fultz已经确定了一个独特的系统，用于解剖一个 急性致死性病毒SIVsmmPBj 14，其分离自猪尾 猕猴接种SIVsmm 9 14个月后，SIVsmm 9是一种原型病毒， 诱发类艾滋病。 申请人将使用连续分离株 在SIVsmm 9接种和恢复之间的定期间隔获得 SIVsmmPBj 14的。 然后，研究人员将尝试定义，在 在生物和分子水平上， 原型和八种体外生物学特性， SIVsmm 9和SIVsmmPBj 14。 这将通过表征 猪尾静脉中SIVsmmPBj 14分离株的致病性 猕猴和八种生物特性中的每一种。 此外一些 将对连续分离株进行分子克隆， 复制分离物的所有表型的病毒， 产生了。 由于SIVsmm 9和 LTR（长末端重复序列）和env（指定病毒）中的SIVsmmPBj 14 膜糖蛋白）和nef（负调节因子[原3' ORF]）基因，申请人提出扩增该基因的相关区域。 通过聚合酶链反应（PCR）从所有分离株的基因组中扩增。 她将 然后对这些序列进行排序， 基因组特定区域的变化。 这应该是 这是可能的，因为已知SIVsmmPBj 14的一些独特性质， 隔离。 基因组准种的体内相关性将是 使用PCR和序列分析直接对从 受感染动物的组织。 最后，为了深入了解 病毒-淋巴细胞相互作用，因为它们与疾病有关，研究者 建议确定SIVsmmPBj 14是否在静息状态下复制， 淋巴细胞，并确定该变体激活的机制 并诱导淋巴细胞增殖。 富尔茨博士会尝试检测 在单细胞水平上感染淋巴细胞，以评估 细胞因子和代谢抑制剂的作用，并确定 通过使用阻断剂诱导增殖的特异性 试剂和富集的细胞群体。

DESCRIPTION: (Adapted from applicant's abstract) To gain a better understanding of the pathogenesis of human immunodeficiency virus (HIV), the investigator proposes to use infection of macaques with a closely related simian virus isolated from sooty mangabey monkeys (SIVsmm). Dr. Fultz has identified a unique system for dissecting the evolution of an acutely lethal virus, SIVsmmPBj14, which was isolated from a pig-tailed macaque 14 months after inoculation with SIVsmm9, a prototype virus that induces AIDS-like disease. The applicant will use sequential isolates obtained at regular intervals between inoculation of SIVsmm9 and recovery of SIVsmmPBj14. The investigator will then attempt to define, at the biological and molecular levels, the evolution of the acutely lethal prototype and eight in vitro biologic properties that differ between SIVsmm9 and SIVsmmPBj14. This will be accomplished by characterization of the sequential SIVsmmPBj14 isolates for pathogenicity in pig-tailed macaques and for each of the eight biologic properties. In addition, some sequential isolates will be molecularly cloned to generate infectious viruses that reproduce all the phenotypes of the isolate from which they were generated. Because of distinct differences between SIVsmm9 and SIVsmmPBj14 in the LTR (long terminal repeat) and env (specifies viral membrane glycoproteins) and nef (negative regulatory factor [formerly 3' ORF]) genes, the applicant proposes to amplify relevant areas of the genome from all isolates by polymerase chain reaction (PCR). She will then sequence these to correlate acquisition of specific biologic properties with changes in specific regions of the genome. This should be possible since some of the unique properties of SIVsmmPBj14 are known to segregate. The in vivo relevance of the genomic quasispecies will be assessed using PCR and sequence analysis directly on DNA isolated from tissues of infected animals. Finally, to gain insight into virus-lymphocyte interactions as they relate to disease, the investigator proposes to determine whether SIVsmmPBj14 replicates in resting lymphocytes and to define the mechanism(s) by which this variant activates and induces lymphocytes to proliferate. Dr. Fultz will attempt to detect infection of lymphocytes at the single cell level, to assess the role of cytokines and the effects of metabolic inhibitors and to determine the specificity of induction of proliferation through the use of blocking agents and enriched cell populations.