Harnessing Tissue Resident Memory T Cells for Immunotherapy of Herpes Virus Infections

项目总结 最近的研究表明，组织驻留记忆(TRM)CD8 T细胞是人类免疫的第一道防线 生殖器皮肤中含有重新激活的HSV-2。对这些独特细胞的深度测序显示了它们的TCR 与随时间采样的同一人的PBMC中检测到的TCR不同，表明它们可能是 针对尚未识别的HSV-2抗原。由于目前所有针对HSV-2的免疫治疗疫苗都是 根据来自PBMC的抗原设计，确定TRM细胞的抗原靶点可能会增强 单纯疱疹病毒免疫治疗新途径的疗效。我们最近确定了α和β的TCR 激光捕获生殖器皮肤活检组织中纯化的TRM链并利用合成生物学创建自体 TCR，并表明在生殖器皮肤中发现的一些常驻CD8 T细胞是HSV-2特异性的，并且是 针对目前HSV-2免疫治疗方法中没有强调的靶点。这项建议利用了 基于单细胞乳剂的条形码技术在高通量无偏识别中的全长 宫颈和外阴CD8T细胞和CD4T细胞的TCRRNA和α链及其蛋白特征 组织，以及HSV-2感染者的PBMC。我们将使用合成生物学来创造“报告者”T细胞 然后可以使用新的HSV-1/HSV-2病毒组来评估其HSV特异性。对这些的识别 TRM人群的同源抗原很可能为疫苗设计提供新的方法。

PROJECT SUMMARY Recent work has shown that tissue resident memory (TRM) CD8+ T cells are the first line of defense for containing reactivated HSV-2 in genital skin. Deep sequencing of these unique cells has shown their TCRs differ from the TCRs detected in PBMC of the same person sampled over time, suggesting they may be directed to as yet unidentified HSV-2 antigens. As all current immunotherapeutic vaccines for HSV-2 are designed based upon antigens derived from PBMC, defining the antigenic targets of TRM cells may enhance the efficacy of novel immunotherapeutic approaches to HSV. We have recently identified TCRs of the α and β chain of laser capture purified TRM from genital skin biopsies and utilized synthetic biology to create autologous TCRs, and shown that some of these resident CD8+ T cells found in genital skin are HSV-2 specific and are directed at targets not currently emphasized in immunotherapeutic approaches to HSV-2. This proposal utilizes single cell emulsion based bar coding technology to identify in a high-throughput unbiased approach full-length TCR α and β chains of CD8+ and CD4+ T cells with RNA/protein signatures of TRM cells in cervical and vulvar tissue, as well as PBMCs of HSV-2 infected persons. We will use synthetic biology to create “reporter” T cells that can then be evaluated for their HSV specificity using a novel HSV-1/HSV-2 vireome. Identification of these cognate antigens of the TRM population is likely to yield new approaches for vaccine design.