HIV Epitope Specific T Cell Responses and Control of HIV Replication

HIV 表位特异性 T 细胞反应和 HIV 复制的控制

• 批准号: 7282098
• 负责人: MARK A JACOBSON
• 金额: $ 23.09万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-15 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7282098
• 关键词: Accounting; Affect; Antigens; Autologous; Biological Assay; Blood; CD3 Antigens; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Count; Cell Degranulation; Cell physiology; Cells; Characteristics; Chronic; Class; Clinical Trials; Dendritic Cells; Development; Disease; Effector Cell; Epitopes; Failure; Flow Cytometry; Frequencies; Future; Gagging; HIV; HIV Infections; HIV vaccine; Haplotypes; Immune; Immune response; Individual; Infection; Infection prevention; Interferon Type II; Interferons; Interleukin-2; Investigation; Measurement; Measures; Monitor; Mutation; Myelogenous; Natural Killer Cells; Numbers; Observational Study; Patients; Pattern; Peptides; Peripheral Blood Mononuclear Cell; Phenotype; Plasma; Proliferating; Proteins; Role; Sampling; Scanning; Specimen; T-Cell Proliferation; T-Lymphocyte; T-Lymphocyte Subsets; TNF gene; Testing; Time; Work; cytokine; experience; immune function; improved; insight; interest; perforin; response

DESCRIPTION (provided by applicant): A major obstacle to the development of an effective HIV vaccine is our lack of understanding of the critical components that constitute a protective, HIV-specific immune response. The aim of this proposal is to investigate, more comprehensively than has been done to date, the potential immunoprotective roles of a broad spectrum of different HIV epitope-specific T cell functions in controlling HIV replication among untreated patients with early HIV infection. The epitope-specific T cell functions that will be measured include CD4+ and CD8+ T cell IFN-g, TNF-a and IL-2 responses (and the maturational state of these responding cells), CD4+ and CD8+ T cell proliferation and CD8+ T cell perforin degranulation. All these functions can be measured by multiparameter flow cytometry after stimulation of PMBC with overlapping peptides that span relevant HIV antigenic proteins. We propose to begin this work by focusing on HIV Gag and Nef epitopes. However, since the measurement of all these T cell responses just these epitopes would require prohibitively large quantities of blood, we will focus on peptide motifs within an individual's autologous Gag/Nef sequence that are known to be HLA-restricted for that individual's haplotype. After sequencing autologous Gag and Nef sequences from stored plasma samples obtained just before untreated patients with early HIV infection established good control of HIV replication, and then again from subsequent timepoints when these same patients have lost that control, we will scan the sequences for HLA haplotype-appropriate motifs and then synthesize them as 9- and 15-mer peptides for Class I and II stimulation, respectively. These peptides will then be used in flow cytometry assays to identify autologous Gag/Nef epitope-specific CD4+ and CD8+ T cell proliferation and IFN-g, TNF-a and IL- 2 responses (and the maturational state of the cytokine-positive cells) and CD8+ T cell perforin degranulation responses. To control for changes in innate immune responses that could also affect control of HIV replication, we will also measure NK cells and function, plasmacytoid and myeloid dendritic cells and regulatory T cells in the same PBMC specimens. Understanding this pattern for clinically important, conserved HIV proteins such as Gag and Nef should improve understanding of the critical components of a protective, HIV-specific immune response and provide insight into the type of HIV epitope-specific T cell responses that may be useful to monitor in future trials of candidate HIV vaccines.

描述（由申请人提供）：开发有效的HIV疫苗的主要障碍是我们对构成保护性的，艾滋病毒特异性免疫反应的关键成分缺乏了解。该提案的目的是比迄今为止更全面地研究，更全面地研究了各种不同HIV表位特异性T细胞功能在控制未经治疗的早期HIV感染患者中的HIV复制中的潜在免疫保护作用。将测量的表位特异性T细胞功能包括CD4+和CD8+ T细胞IFN-G，TNF-A和IL-2响应（以及这些响应细胞的成熟态），CD4+和CD8+ T细胞增殖以及CD8+ T细胞perforin脱粒。所有这些功能都可以通过跨越相关HIV抗原蛋白的重叠肽刺激PMBC后通过多参数流式细胞仪来测量。我们建议通过关注艾滋病毒插科打g和nef表位来开始这项工作。但是，由于仅这些表位的所有这些T细胞反应的测量将需要大量的血液，因此我们将重点放在个体的自体GAG/NEF序列中的肽基序上，这些肽序列已知对该个体的单倍型受到HLA的限制。 After sequencing autologous Gag and Nef sequences from stored plasma samples obtained just before untreated patients with early HIV infection established good control of HIV replication, and then again from subsequent timepoints when these same patients have lost that control, we will scan the sequences for HLA haplotype-appropriate motifs and then synthesize them as 9- and 15-mer peptides for Class I and II stimulation, respectively.然后，这些肽将用于流式细胞仪测定法，以鉴定自体GAG/NEF表位特异性CD4+和CD8+ T细胞增殖，以及IFN-G，TNF-A和IL-2反应（以及细胞因子阳性细胞的成熟状态）和CD8+ T细胞透明蛋白脱糖蛋白脱粒反应。为了控制可能影响对HIV复制的控制的先天免疫反应的变化，我们还将测量NK细胞和功能，浆细胞类动物和髓样树突状细胞以及相同PBMC标本中的调节性T细胞。了解这种模式的临床重要，保守的HIV蛋白（例如GAG和NEF）应提高对保护性，HIV特异性免疫反应的关键成分的理解，并提供对HIV表位特异性T细胞反应类型的洞察力，这些反应可能可用于监测候选HIV疫苗的未来试验中可能有用。