Impact of HIV-1 Fitness on Disease Progression

描述（由申请人提供）：本提案的基本目标将 是为了测量来自感染个体的HIV-1分离物的体外适应性， 进化枝B和非进化枝B HIV-1，然后将这种适应性与各种 疾病进展的预测因子。一项初步研究显示， HIV-1适应性和病毒载量之间的关系。为了扩大这项研究，多个 从大约80名艾滋病毒感染者中分离出的HIV- 1已被 从瑞典卡罗林斯卡研究所获得， 比利时安特卫普的热带医学。在具体目标1中，我们将竞争每一个 患者分离株与四种对照HIV-1菌株，以确定相对离体 适合度值然后将这些适应度值与各种临床 参数如CD 4细胞计数和病毒载量。这一目标的目的是 为了扩展先前的一项试点研究，该研究显示， 健康和疾病进展。最终， 进展可能在HIV-1分离株或亚型的分布中起作用， 疫具体目标2是比较内部和 亚型间适应度来自进化枝A、B、C和 D通过外周血单个核细胞的成对竞争。初步 数据表明，不同亚型的分离株之间的差异大于 相同的亚型。因此，在具体目标3中，我们将研究是否可能 亚型适应性的差异可能影响非进化枝B的疾病进展 感染.如在目的1中一样，来自HIV-I的原代HIV-1分离株的离体适合性是由以下因素决定的： 大约150名感染艾滋病毒的乌干达人和津巴布韦人（参加一个 NICHD资助的研究）将与疾病的各种预测因子进行比较 进展由于病毒首先从血液和阴道中获得， 在原发性感染的分泌物，我们也将进行亚型间 使用Langerhan细胞的竞争实验。适合度的病毒繁殖 研究是一种费力的技术，可以通过实验来规避。 具体目标4。使用测定来查看在该方法中的多个步骤。 HIV-1复制周期在双重病毒感染，我们现在有初步的 数据表明，HIV-1的进入可能正在控制竞争。我们将 在SPECIFIC AIM 4中证实了这一初步观察结果，然后使用一种新的 产生用env基因假型化的HIV- 1 NL 4 -3克隆的克隆策略 直接从患者样本中提取。这些env假型HIV-1之间的竞争 然后将克隆与两个繁殖的HIV-1之间的竞争进行比较 两个病人样本的分离株

DESCRIPTION (provided by applicant): The basic objective of this proposal will be to measure the ex vivo fitness of HIV-1 isolates from individuals infected with clade B and non-clade B HIV-1 and then compare this fitness to various predictors of disease progression. A pilot study revealed a strong correlation between ex vivo HIV-1 fitness and viral load. To extend this study, multiple HIV- 1 isolates from approximately 80 HIV-infected individuals have been obtained from the Karolinska Insititute in Sweden and at the Institute of Tropical Medicine in Antwerp, Belgium. In SPECIFIC AIM1, we will compete each patient isolate with four control HIV-1 strains to determine a relative ex vivo fitness value. These fitness values will then be compared to various clinical parameters such as CD4 cell counts and viral load. The objective of this aim is to extend a previous pilot study showing a strong correlation between HIV-1 fitness and disease progression. Ultimately the rate of intrapatient disease progression may play a role in distribution of HIV-1 isolates or subtypes in the epidemic. SPECIFIC AIM 2 is to compare differences in intra- and intersubtype fitness. The fitness of multiple isolates from clades A, B, C, and D via pair-wise competitions in peripheral blood mononuclear cells. Preliminary data suggests a greater difference between isolates of different subtypes than of the same subtype. Thus, in SPECIFIC AIM 3, we will investigate if possible differences in subtype fitness may affect disease progression in non-clade B infections. As in aim 1, the ex vivo fitness of primary HIV-1 isolates from approximately 150 HIV-infected Ugandans and Zimbabweans (participating in an NICHD-funded study) will be compared to various predictors of disease progression. Since virus will be first obtained from both blood and vaginal secretions during primary infection, we will also perform intersubtype competition experiments using Langerhan cells. Viral propagation for fitness studies is a laborious technique that may be circumvented with the experiments proposed in SPECIFIC AIM 4. Using an assay to look at multiple steps in the HIV-1 replication cycle during dual virus infections, we now have preliminary data to suggest that HIV-1 entry may be controlling the competition. We will confirm this preliminary observation in SPECIFIC AIM 4 and then use a novel cloning strategy to produce HIV- 1 NL4-3 clones pseudotyped with env genes directly from patient samples. Competitions between these env pseudotyped HIV-1 clones will then be compared to competitions between two propagated HIV-1 isolates from the same two patient samples.