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 ? and ? 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 ? and ? 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.
This proposal is directed at extending the observations that tissue resident memory (TRM) CD8+ T cells to HSV-2 are the first line of defense for containing reactivated HSV-2 in genital skin. The antigenic targets of these as yet uncultivable cells are currently undefined. Identifying these and including them in an immunotherapeutic vaccine would likely enhance potential efficacy of an HSV-2 vaccine. We will identify and construct HSV TRM cells from genital skin and cervix, and compare their antigenic specificity and functional characteristics to PBMC derived T cells. These studies will help define if the TRM system can be harnessed to treat/cure chronic viral infections.
