PHENOTYPE AND GENOTYPE OF HIV-1 STRAINS FROM CHILDREN

出现艾滋病毒相关疾病症状的年龄存在很大差异 首先出现在围产期感染的儿童中。在长期幸存者中 特定的先天病毒学特性和宿主免疫的结合 与病毒学相比，反应可以使感染“受到控制” 其他快速进展的受感染婴儿的参数 疾病。 我们将详细说明生物之间存在的关系 原发儿科分离株表现出的特征（病毒表型） HIV-1 的体外检测和疾病的临床表现。我们会 确定连续分离株是否变得更具致病性 感染进展以及儿科长期幸存者是否携带病毒 显示出与致病性降低相关的表型的分离株。 我们将尝试在单核细胞来源的巨噬细胞中培养所有分离株 了解这种增长特性如何演变以及它是否与 临床疾病或中枢神经系统疾病的进展发生变化。我们会 确定特定之间是否存在一致的关联 env 和病毒表型的 V1、V2 和 V3 环中的序列。我们会 还研究了病毒分离株的基因型在体内传代的程度 培养物符合野生病毒株的基因型 从血液单核细胞和血浆中从头分离。为了 更详细地研究基因型和表型之间的关系， 少数母亲和儿童的传染性原病毒 DNA 克隆 将获得分离株并检查其生物学行为。部分 这些克隆的 env 区域将被测序。我们会寻找 与特定表型相关的遗传特征。 在这些研究中，我们将广泛使用样本档案 （病毒储液、培养上清液、冷冻 PBMC、血浆等） 从康涅狄格州 70 名 HIV-1 感染儿童身上收集， 包括 27 名长期幸存者，以及我们将要收集的新标本 前瞻性地收集。我们的大多数患者也参加了 前瞻性、纵向队列研究；因此，人口统计学和临床 数据可用于定义表型、基因型和 疾病的临床表现。

There is great variation in the age when signs of HIV-associated illness first appear in children infected perinatally. In long-term survivors a combination of specific, innate virologic properties and host immune responses keep the infection "in check", as compared to virologic parameters in other infected infants who develop rapidly progressive disease. We will detail the relationships that exist between the biologic characteristics (viral phenotype) displayed by primary pediatric isolates of HIV-1 in vitro and clinical manifestations of disease. We will determine whether sequential isolates become more pathogenic as the infection progresses and whether pediatric long-term survivors have viral isolates which display phenotypes associated with decreased pathogenicity. We will attempt to cultivate all isolates in monocyte-derived macrophages to learn how this growth property evolves and if it correlates with changes in the pace of clinical illness or with CNS disease. We will determine whether there are consistent associations between specific sequences in the V1, V2 and V3 loops of env and viral phenotype. We will also study the degree to which the genotype of virus isolates passaged in culture conforms with the genotype of feral virus strains that are isolated de novo from blood mononuclear cells and plasma. In order to study in greater detail the relationships between genotype and phenotype, infectious proviral DNA clones of a small number of mother and child isolates will be obtained and their biologic behavior examined. Portions of the env region of these clones will be sequenced. We will look for genetic features that are associated with particular phenotypes. In these studies we will make extensive use of an archive of samples (virus stocks, culture supernatant fluids, frozen PBMC, plasma, etc) that have been collected from 70 HIV-1 infected children in Connecticut, including 27 long-term survivors, and of new specimens that we will collect prospectively. Most of our patients are also enrolled in a prospective, longitudinal cohort study; hence, demographic and clinical data are available to define associations between phenotype, genotype and clinical expression of disease.