PHENOTYPE AND GENOTYPE OF HIV-1 STRAINS FROM CHILDREN

儿童 HIV-1 菌株的表型和基因型

• 批准号: 2457851
• 负责人: WARREN A ANDIMAN
• 金额: $ 29.07万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2457851
• 关键词: child (0-11); clinical research; genetic strain; genotype; human immunodeficiency virus 1; human subject; molecular cloning; nucleic acid sequence; pathologic process; pediatric AIDS; phenotype; tissue /cell culture; virulence; virus classification

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.

出现艾滋病毒相关疾病迹象的年龄差异很大， 首先出现在围产期感染的儿童中。在长期存活者中， 特异性、先天病毒学特性和宿主免疫的组合 与病毒学相比， 其他感染婴儿中的参数， 疾病 我们将详细说明生物学之间存在的关系， 主要儿科分离株显示的特征（病毒表型） HIV-1在体外和疾病的临床表现。我们将 确定连续分离株是否变得更具致病性， 感染进展和儿科长期存活者是否有病毒 分离物显示与降低的致病性相关的表型。 我们将尝试在单核细胞衍生的巨噬细胞中培养所有分离株 来了解这种生长特性是如何演变的， 临床疾病或CNS疾病的速度变化。我们将 确定是否存在特定 env的V1、V2和V3环中的序列和病毒表型。我们将 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重新分离。为了 更详细地研究基因型和表型之间的关系， 少数母婴感染性前病毒DNA克隆 将获得分离株并检查其生物学行为。部分 将对这些克隆的env区进行测序。我们将寻找 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 （病毒原液、培养上清液、冷冻PBMC、血浆等）， 从康涅狄格州的70名HIV-1感染儿童身上收集， 包括27个长期存活者，以及新的标本， 前瞻性地收集。我们的大多数病人也参加了一个 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。