HIV VARIATION AND CROSS CLADE CTL RECOGNITION

描述：(改编自申请者摘要)本项目响应 计划公告#PA-98-011“艾滋病毒变异对免疫学的影响” 这项建议的目的是调查跨领域的艾滋病毒 特异性CTL应答。广谱反应性HIV的刺激作用 特异性CTL反应被认为是成功的HIV疫苗的关键。这个 高度的序列变异是HIV感染的一个显著特征，以及 这是疫苗开发的主要障碍。然而，很少有信息是 了解国际不同人群中HIV特异性CTL反应 人类白细胞抗原亚型，感染HIV分支变异体。通过我们的合作者，我们拥有 获取非洲艾滋病毒感染者的PBMC样本，中国，南方 美国和泰国感染了HIV-1和HIV-2。在第一个具体目标中， 我们将确定交叉反应性CTL表位是否存在于 感染不同HIV-1分支的人群。我们将测量十字 用灵敏的ELISPOT方法反应CTL反应，使用一组 表达HIV-1基因产物的树枝状重组痘苗病毒 Env、GAG、P01和NEF，以及分支特异性多肽。批量培养CTL反应 将通过多克隆和抗原特异性的重新刺激产生并测试 自体或配对的B淋巴母细胞靶点与支系特异性 抗原。我们将把所有人的CTL频率与病毒RNA的血浆载量联系起来 并评估氨基酸是否在最佳定义的最小范围内发生变化 表位可以使HIV CTL表位更具交叉反应。在第二个具体例子中 目的：确定HLV~2感染是否刺激交叉的CTL 与HIV-1反应或反之亦然，以及一些人是否受到 HIV-1的一个分支对抗另一个分支的重叠感染。

DESCRIPTION: (Adapted from Applicant's Abstract) This project is responsive to the Program Announcement # PA-98-011 "Impact of HIV Variation on Immunological Recognition." The aim of this proposal is to investigate cross clade HIV specific CTL responses. Stimulation of a broad cross clade reactive HIV specific CTL response is considered crucial for a successful HIV vaccine. The high degree of sequence variation is a prominent feature of HIV infection, and a major impediment to vaccine development. However, very little information is known about HIV specific CTL responses in international populations of diverse HLA types, infected with HIV clade variants. Through our collaborators, we have access to PBMC samples from HIV infected individuals in Africa, China, South America and Thailand infected with HIV-1 and HIV-2. In the first specific aim, we will determine whether cross-reactive CTL epitopes exist in diverse populations infected with different clades of HIV-1. We will measure cross reactive CTL responses with a sensitive ELISPOT assay, using both a panel of clade specific recombinant vaccinia viruses expressing HIV-1 gene products from env, gag, p01 and nef, and clade specific peptides. Bulk culture CTL responses will be generated with polyclonal and antigen specific restimulation and tested on autologous or matched B lymphoblastoid cell targets with clade specific antigens. We will correlate CTL frequency with plasma load of viral RNA for all clades and assess whether amino acid changes in optimally defined minimal epitopes can make HIV CTL epitopes more cross reactive. In the second specific aim, we will ascertain whether infection with HlV~2 stimulates CTL which cross react with HIV-1 or vice versa, and whether some individuals are protected by one clade of HIV-1 against super-infection with another.