Project description:We conducted immune- and RNA-sequencing of HLA-A24-restricted CMVpp65-specific CTLs to better understand the immune reconstitution of CMV-CTLs after allo-HCT. To the best of our knowledge, this is the first report on the features of TCRβ-CDR3, diversity, and GEP of HLA-A24 CMV-CTLs according to the CMV-reactivation pattern among recipients after allo-HCT. In addition, we further sought to demonstrate homogeneity or heterogeneity according to individual CTL clones using single-cell RNA-sequencing technology.
Project description:We conducted immune- and RNA-sequencing of HLA-A24-restricted CMVpp65-specific CTLs to better understand the immune reconstitution of CMV-CTLs after allo-HCT. To the best of our knowledge, this is the first report on the features of TCRβ-CDR3, diversity, and GEP of HLA-A24 CMV-CTLs according to the CMV-reactivation pattern among recipients after allo-HCT. In addition, we further sought to demonstrate homogeneity or heterogeneity according to individual CTL clones using single-cell RNA-sequencing technology.
Project description:The goal of this study is to understand the transcriptional features of CD4+ memory T cells induced by CMV seropositivity and explore their TCR repertoire clonality.
Project description:The latent human Cytomegalovirus (hCMV) infection can pose a serious threat of reactivation and disease occurrence in immune-compromised individuals, as well as burdens the immune system in immune-competent individuals. Though, T cells are at the core of the protective immune response to hCMV infection, a detailed characterization of different T cell subsets involved in protection against the hCMV infection is lacking. Here we analyzed the single-cell transcriptomes and the single-cell T cell antigen receptor (TCR) repertoires of over 8000 hCMV-reactive peripheral T cells isolated from different memory compartments. The hCMV-reactive T cells were highly heterogeneous and consisted of different developmental memory and functional T cell subsets such as, the long-term memory precursors and effectors, T helper-17, T regulatory cells (TREGs) and cytotoxic T lymphocytes (CTLs). The hCMV-antigen specific TREGs were enriched for molecules linked to their suppressive function and interferon response genes. The CTLs were of two types, the pre-effector and effector like. Of particular interest was the mixture of both CD4-CTLs and CD8-CTLs in both the pre-effector and effector cytotoxic clusters, suggesting that both CD4-CTLs and CD8-CTLs share transcriptomic signatures. The huge TCR clonal expansion of both the cytotoxic clusters imply their predominant role in protective immune response to CMV. Further the clonotype sharing between the CTL clusters and the long-term memory clusters, indicate potential progenitors of CD4-CTLs. Together our study has identified many subsets of hCMV-specific memory T cells that may have implication in better understanding the hCMV-specific T cell immunity to design vaccination strategies and therapeutics.
Project description:<p>High-throughput linking of T cell receptor (TCR) sequences to their binding antigens is vital for immune profiling, yet challenging. We present Tetramer associated TCR Sequencing (TetTCR-Seq) to address this challenge. Binding is determined using a library of DNA-barcoded antigen tetramers that are rapidly and inexpensively generated using an in vitro transcription/translation platform. We included CMV+ donors (CMV seropositive donors who are infected with Cytomegalovirus) to screen for CMV specific TCRs.</p>
