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

项目摘要 最近的研究表明，组织驻留记忆（TRM）CD 8 + T细胞是免疫缺陷的第一道防线。 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， 与随时间采样的同一个人的PBMC中检测到的TCR不同，这表明它们可能是 针对尚未鉴定的HSV-2抗原。由于目前所有的HSV-2免疫疫苗都是 基于来源于PBMC的抗原设计，定义TRM细胞的抗原靶标可以增强TRM细胞的免疫原性。 新的免疫治疗方法对HSV的有效性。我们最近发现了α和β受体 激光捕获链从生殖器皮肤活检中纯化TRM，并利用合成生物学来创建自体 TCR，并表明在生殖器皮肤中发现的这些常驻CD 8 + T细胞中的一些是HSV-2特异性的， 针对目前在HSV-2的免疫治疗方法中未强调的靶点。该提案利用 基于单细胞乳剂的条形码技术，以高通量无偏方法鉴定全长 宫颈和外阴中CD 8+和CD 4 + T细胞的TCR α和β链与TRM细胞的RNA/蛋白质特征 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”T细胞 然后可以使用新的HSV-1/HSV-2病毒组来评估它们的HSV特异性。识别这些 TRM群体的同源抗原可能产生用于疫苗设计的新方法。

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.