Single-cell transcriptomic and T cell antigen receptor analysis of human cytomegalovirus (hCMV)-specific memory T cells reveals effectors and pre-effectors of CD8+- and CD4+-cytotoxic T cells.

Latent human cytomegalovirus (hCMV) infection can pose a serious threat of reactivation and disease occurrence in immune-compromised individuals. Although T cells are at the core of the protective immune response to hCMV infection, a detailed characterization of different T cell subsets involved in hCMV immunity is lacking. Here, in an unbiased manner, we characterized over 8000 hCMV-reactive peripheral memory T cells isolated from seropositive human donors, at a single-cell resolution by analysing their single-cell transcriptomes paired with the T cell antigen receptor (TCR) repertoires. The hCMV-reactive T cells were highly heterogeneous and consisted of different developmental and functional memory T cell subsets such as, long-term memory precursors and effectors, T helper-17, T regulatory cells (TREGs) and cytotoxic T lymphocytes (CTLs) of both CD4 and CD8 origin. The hCMV-specific TREGs, in addition to being enriched for molecules known for their suppressive functions, showed enrichment for the interferon response signature gene sets. The hCMV-specific CTLs were of two types, the pre-effector- and effector-like. The co-clustering of hCMV-specific CD4-CTLs and CD8-CTLs in both pre-effector as well as effector clusters suggest shared transcriptomic signatures between them. The huge TCR clonal expansion of cytotoxic clusters suggests a dominant role in the protective immune response to CMV. The study uncovers the heterogeneity in the hCMV-specific memory T cells revealing many functional subsets with potential implications in better understanding of hCMV-specific T cell immunity. The data presented can serve as a knowledge base for designing vaccines and therapeutics.

Single-cell characterisation of tissue homing CD4 + and CD8 + T cell clones in immune-mediated refractory arthritis.

BACKGROUND: Immune-mediated arthritis is a group of autoinflammatory diseases, where the patient's own immune system attacks and destroys synovial joints. Sustained remission is not always achieved with available immunosuppressive treatments, warranting more detailed studies of T cell responses that perpetuate synovial inflammation in treatment-refractory patients. METHODS: In this study, we investigated CD4 + and CD8 + T lymphocytes from the synovial tissue and peripheral blood of patients with treatment-resistant immune-mediated arthritis using paired single-cell RNA and TCR-sequencing. To gain insights into the trafficking of clonal families, we compared the phenotypes of clones with the exact same TCRß amino acid sequence between the two tissues. RESULTS: Our results show that both CD4 + and CD8 + T cells display a more activated and inflamed phenotype in the synovial tissue compared to peripheral blood both at the population level and within individual T cell families. Furthermore, we found that both cell subtypes exhibited clonal expansion in the synovial tissue. CONCLUSIONS: Our findings suggest that the local environment in the synovium drives the proliferation of activated cytotoxic T cells, and both CD4 + and CD8 + T cells may contribute to tissue destruction and disease pathogenesis.

T cell receptor repertoire deciphers anti-tuberculosis immunity.

T cell induced cellular immunity is considered to be extremely important for the control of tuberculosis (TB). T cell receptor (TCR), the key component responsible for the specificity and clustering of T cells, holds the potential to advance our understanding of T cell immunity against TB infection. This review systematically expounded the study progressions made in the field of TB-relevant TCRs based on single cell sequencing together with GLIPH2 technology and initiated a comparison of the T cell distribution between peripheral blood and infected organs. We divided clonal expanded T cell clones into recirculation subsets and local subsets to summarize their distinctions in clonal abundance, TCR sequences and antigenic specificity. Notably, local expansion appears to drive the primary variances in T cell subsets between these two contexts, indicating the necessity for further exploration into the functions and specificity of local subsets.

scRNA + TCR-seq revealed the dual TCR pTh17 and Treg T cells involvement in autoimmune response in ankylosing spondylitis.

OBJECTIVE: Th17 and Treg play important roles in AS, but their single and dual TCR pairing types, ratios, and CDR3 characteristics remain unknown. METHODS: Single-cell RNA + TCR-seq results from six AS patients were used to cluster T-cell subpopulations and analyze the single and dual TCR T cell ratio, diversity/clonality/overlap of CDR3, and expression of transcription factors. RESULTS: 1. AS patients have about 10% of dual TCR T cells, and SFMC have decreased diversity CDR3 libraries and significant clonal proliferation compared to PBMC. 2. Dual TCR ratio: memory T > naive T; pTh 17 > Th17; Treg /Th17/Th1/EM significantly higher than naive CD4 + T/CM, Pathogenic Th17 cells contain clonally proliferating single TCR and dual TCR cells. 3. The expression of single TCR and dual TCR transcription factors of each T cell subpopulation was basically the same, but there was differential expression of characteristic transcription factors, e.g. Foxp3, CTLA4, STAT5B, IL10RB, LAG3 in dual TCR Treg was higher than that of single TCR Treg; TNFSF10/12, TNFRSF4/14, CCL5, KLRB1 in dual TCR pTh17 were significantly higher than those in single TCR pTh17. 4. Between naive CD4 + T, pTh17, Th1 and Treg, there are partially identity identical tcr paired cells. CONCLUSIONS: The high proportion of dual TCR T cells such as pTh17 and Treg in AS and the high expression of some transcription factors suggested a close association with self-response in AS; The overlap of CDR3 between Th1, Th17,pTh17, and Treg in AS suggested that the subpopulations may be differentiated from each other to regulate the inflammatory homeostasis and progression.