HSV-2 specificity and phenotyping of tissue-based T cells in genital skin biopsies of HSV-2 reactivation

HSV-2 重新激活的生殖器皮肤活检中基于组织的 T 细胞的 HSV-2 特异性和表型

• 批准号: 10092943
• 负责人: Emily Ford
• 金额: $ 18.93万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092943
• 关键词: Affect; Animal Model; Antigenic Specificity; Antigens; Area; Binding Sites; Biopsy; Blood; Blood Cells; CD4 Positive T Lymphocytes; Cell physiology; Cells; Chronic; Clinical; Containment; Development; Disease; Dose; Effectiveness; Epidemic; Epitope Mapping; Epitopes; Future; Gene Expression Profile; Gene Expression Profiling; Generations; Genetic Transcription; Genital; Genitalia; Goals; HIV; HIV-1; Health Benefit; Herpes Simplex Virus Vaccines; Histologic; Human; Human Herpesvirus 2; Immune response; Immune system; Immunocompetent; Immunocompromised Host; Immunotherapeutic agent; In Situ Hybridization; Individual; Infection; Investigation; K-Series Research Career Programs; Laboratories; Lead; Learning; Location; Mentors; Molecular; Morbidity - disease rate; Natural History; Peripheral Blood Mononuclear Cell; Peripheral Nerves; Persons; Phase; Phenotype; Play; Population; Principal Investigator; Production; Public Health; RNA; Research; Research Personnel; Risk; Role; Sampling; Series; Simplexvirus; Site; Skin; Specificity; T cell receptor repertoire sequencing; T memory cell; T-Cell Immunologic Specificity; T-Cell Receptor; T-Lymphocyte; T-cell receptor repertoire; Techniques; Technology; Testing; Time; Tissue Expansion; Tissues; Ulcer; Vaccinated; Vaccination; Vaccines; Viral; Viral Antigens; Work; arm; base; career; cytokine; design; educational atmosphere; experience; fighting; first responder; foot; global health; improved; lens; optimism; programs; public health relevance; repository; transcriptomics; transmission process; vaccine candidate; vaccine development; vaccine efficacy; vaccine trial; virology

PROJECT SUMMARY / ABSTRACT Summary of Proposal: This proposal supports a 5-year mentored career development award investigating the tissue-based immune response through the lens of a recent phase 1 immunotherapeutic vaccine trial with the long-term goal of contributing to the development of an effective herpes simplex virus-2 (HSV-2) vaccine and improving global health.. This trial involved three sequential doses of an immunotherapeutic vaccine, HSV529, in individuals with symptomatic HSV-2 infection. T-cell receptor (TCR) repertoire analysis was performed of T cells in small biopsies of the area of HSV-2 reactivation in comparison to T cells from biopsies of uninvolved skin and HSV-2 reactive CD4+ T cells from peripheral blood to research two main lines of inquiry. The first is the interrelatedness, over time, of the TCR repertoire of the genital skin at HSV-2 reactivation sites, the arm, and HSV-2 specific CD4+ T cells over time; whether T cells detected in genital tissue are detectable in blood with standard sampling techniques. The second is how these relationships and dynamics change over the course of an immunotherapeutic vaccination. Preliminary findings suggest that some T cells increased in number after vaccination in everyone, with very little overlap between the skin and the blood. The central hypothesis is that T cells that respond to a vaccine targeting HSV-2 are likely to be HSV-2-specific, and that T cells seen in tissue at high copy number over a long period of time are likely to be tissue residents. The objective of this proposal is to learn more about these resident T cells, which are otherwise difficult to study. The rationale for this is that vaccine development for HSV-2 has thus far not been successful, and that because there is so little overlap between the skin and the blood, that looking directly at the tissue may help identify a viral antigen or a way to target skin-based T cells to improve vaccine efficacy. Research plan: AIM 1a will identify the specificity of T cells of that were seen most frequently in the region of HSV-2 reactivation, focusing on T cells that expanded in number after immunotherapeutic vaccination, as these are the most likely to be herpes-specific, tissue-resident cells. AIM 1b focuses on establishing the spatial location of these cells in small sections of biopsies using fluorescent molecular tags to visualize their arrangement in skin. AIM 2 focuses on learning more about these T cells by using transcriptional profiling, a technique that separates each of the T cells in a biopsy and looks at them one by one to uncover the cellular processes that define the activity of each cell. We will incorporate information from TCR repertoire analysis to interpret the results of transcriptional profiling more effectively and accurately. This approach uses small skin samples in the most comprehensive manner available to describe the fight between HSV-2 and the human immune system. By conducting this research with an extremely experienced and uniquely skilled group of mentors in the rich educational environment of the FHCRC and UW, Dr. Ford will become an effective and productive lead investigator.

项目摘要 /摘要 提案摘要：该提案支持一项为期5年的指导职业发展奖，调查了 通过最近的1期免疫治疗疫苗试验的晶状体基于组织的免疫反应 为开发有效的单纯疱疹病毒-2（HSV-2）疫苗的发展的长期目标 改善全球健康。该试验涉及三种顺序剂量的免疫治疗疫苗HSV529， 有症状性HSV-2感染的人。对T进行T细胞受体（TCR）曲目分析 与未参与皮肤活检的T细胞相比 HSV-2反应性CD4+ T细胞从外周血到研究两个主要询问线。第一个是 随着时间的流逝，HSV-2重新激活部位，手臂和 随着时间的推移，HSV-2特异性CD4+ T细胞；在生殖器组织中检测到的T细胞是否可以在血液中检测到 标准抽样技术。第二是这些关系和动态在整个过程中的变化 免疫治疗疫苗。初步发现表明，某些T细胞在 每个人的疫苗接种，皮肤和血液之间几乎没有重叠。中心假设是 对靶向HSV-2疫苗反应的T细胞可能是HSV-2特异性的，并且在组织中看到的T细胞 长时间内，高拷贝数可能是组织居民。该提议的目的 是为了更多地了解这些居民T细胞，否则很难研究。这样做的理由是 到目前为止，HSV-2的疫苗开发尚未成功，并且因为重叠很少 在皮肤和血液之间，直接看组织可能有助于识别病毒抗原或一种方法 靶向皮肤的T细胞可提高疫苗功效。研究计划：AIM 1A将确定T的特异性 在HSV-2重新激活的区域中，最频繁地看到细胞，重点是扩展的T细胞 免疫治疗疫苗接种后的数字，因为这些是最有可能是疱疹特异性组织居民 细胞。 AIM 1B专注于使用使用 荧光分子标签可视化它们在皮肤中的排列。 AIM 2专注于了解有关这些t的更多信息 通过使用转录分析，该细胞是一种在活检中分开每个T细胞并查看的技术 它们一个一个一个揭示定义每个细胞活性的细胞过程。我们将合并 来自TCR曲目分析的信息，以更有效地解释转录分析的结果 准确。这种方法以最全面的方式使用小型皮肤样品来描述 HSV-2与人类免疫系统之间的战斗。通过经验丰富的 在FHCRC和UW丰富的教育环境中，熟练的指导者独特的团队，福特博士将 成为有效且富有成效的研究员。