Molecular Biology

全球流行的HIV-1毒株具有极大的变异性，这种多样性对艾滋病疫苗的开发构成了主要障碍。这个HIVRAD小组将测试来自HIV-1共识和/或祖先序列的免疫原是否能够引起比来自当代HIV-1毒株的免疫原更广泛的交叉反应免疫反应。在这种背景下，一个重要的目标是从代表多种不同亚型的急性/早期HIV-1感染中产生全长基因组序列以及功能性env/rev表达盒。这些试剂将用于检查最近传播的HIV-1毒株是否表现出独特的序列特征和/或生物学特性，这些特性应该被纳入艾滋病疫苗设计。最近传播的当代HIV-1 毒株也是最适合用来测试疫苗引起中和抗体反应的效力和广度的病毒。来自急性/早期HIV-1感染的功能性包膜克隆将被用来产生这样的病毒面板。分子生物学核心将通过对每年30至40例急性/早期HIV-1感染的分子特征来支持这一HIVRAD努力。具体目标包括： 1.从急性或早期HIV-1感染患者中扩增、克隆和测序代表广泛M组亚型的准种gp160(env/rev)全长基因。 2.从这些患者中获得最近传播的HIV-1的全长基因组序列。 3.从急性/早期HIV-1感染中产生有功能的env克隆，以产生 用于中和研究的当代亚型特异性控制基因和伪病毒面板。 4.为整个HIVRAD团队提供一般分子生物学服务。 由此产生的序列和试剂将对所有其他艾滋病毒/艾滋病项目的成功至关重要，因为它们将促进产生(I)急性/早期艾滋病毒-1数据库(项目#1)；(Ii)当代功能性环境控制基因(项目#2、#3和#4)，以及(Iii)一组亚型特异的当代急性/早期艾滋病毒-1毒株，以测试集中免疫原引发的中和抗体反应的广度和效力(项目#2和中和核心C)。

Globally circulating strains of HIV-1 are extraordinarily variable, and this diversity poses a major obstacle to AIDS vaccine development. This HIVRAD team will test whether immunogens derived from HIV-1 consensus and/or ancestral sequences are capable of eliciting more broadly cross-reactive immune responses than immunogens derived from contemporary HIV-1 strains. In this context, an important objective is to generate full-length genome sequences as well as functional env/rev expression cassettes from acute/early HIV-1 infections representative of multiple different subtypes. These reagents will be used to examine whether recently transmitted HIV-1 strains exhibit unique sequence signatures and/or biological properties that should be incorporated into AIDS vaccine design. Recently transmitted, contemporary HIV-1 strains are also the most appropriate viruses against which to test the potency and breadth of vaccine elicited neutralizing antibody responses. Functional env clones from acute/early HIV-1 infections will be used to generate such virus panels. The Molecular Biology Core will support this HIVRAD effort by molecularly characterizing 30 to 40 acute/early HIV-1 infections per year. Specific aims include: 1. To PCR amplify, clone and sequence full-length gp160 (env/rev) quasispecies from patients with acute or early HIV-1 infection representing a broad range of group M subtypes. 2. To derive full-length genome sequences of recently transmitted HIV-1 from these same patients. 3. To generate functional env clones from acute/early HIV-1 infections for the generation of contemporary subtype specific control genes and pseudovirus panels for neutralization studies. 4. To provide general molecular biology services to the entire HIVRAD team. The resulting sequences and reagents will be critical for the success of all other HIVRAD projects since they will facilitate the generation of (i) an acute/early HIV-1 database (Project #1); (ii) functional contemporary env control genes (Projects #2, #3 and #4), and (iii) a panel of subtype specific, contemporary acute/early HIV-1 strains to test the breadth and potency of neutralizing antibody responses elicited by centralized immunogens (Project #2 and Neutralization Core C).