Immune profiling of γδ T cells after human intestinal transplantation

人肠道移植后 γδ T 细胞的免疫分析

• 批准号: 10351494
• 负责人: Jianing Fu
• 金额: $ 24.3万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10351494
• 关键词: Age; Allografting; Antigen-Presenting Cells; Antigens; Attenuated; B-Cell Antigen Receptor; Blood Circulation; Bone Marrow; Bone Marrow Cytometry; CD8B1 gene; Cells; Chimerism; Clinical; Clone Cells; Cytometry; Data; Development; Engraftment; Equilibrium; Eragrostis; Gastrointestinal tract structure; Gene Expression Profile; Goals; Graft Rejection; Hematopoietic System; Hematopoietic stem cells; High-Throughput Nucleotide Sequencing; Host Defense; Host vs Graft Reaction; Human; Immune; Immune Tolerance; Immune response; Immunologic Surveillance; Infection; Intestines; Investigation; Journals; Life; Link; Liver; Lymphocyte; Mediating; Mixed Lymphocyte Culture Test; Mucous Membrane; Organ; Organ Transplantation; Outcome; Pattern; Phenotype; Physiological; Privatization; Procedures; Progenitor Cell Engraftment; Property; Quality of life; Recording of previous events; Research; Rodent; Role; Savings; Shapes; T memory cell; T-Cell Receptor; T-Lymphocyte; T-cell receptor repertoire; Technology; Testing; Tissues; Transplant Recipients; Transplantation; Work; allograft rejection; clinical investigation; complementarity-determining region 3; cytotoxic; effector T cell; gastrointestinal transplantation; genomic platform; improved; insight; next generation sequencing; novel; novel strategies; pathogen; peripheral blood; reconstitution; response; single-cell RNA sequencing; trafficking; γδ T cells

PROJECT SUMMARY γδ T cells can recognize diverse antigens and exert disparate functions. The innate- and adaptive-like features of human γδ T cells may be driven by differential γδ T cell receptor (TCR) repertoires, which can be shaped by tissue compartmentalization, age and history of antigen exposure. Despite comprising a significant proportion of resident T cells in many organs, including gut and liver, γδ T cells and their possible role in transplantation outcomes are largely under‐researched. Our overarching goals are to elucidate the fundamental mechanisms of how γδ T cells participate in allograft rejection and host defense after human intestinal transplantation (ITx). We revealed that clinical outcomes after ITx are largely determined by the balance between graft-versus-host (GvH) and host-versus-graft (HvG) alloreactivities, using an approach that identifies alloreactive T cell clones following pre-Tx mixed lymphocyte reactions on the basis of TCRβ CDR3 high throughput sequencing (TCRβ-seq). We showed that GvH-reactive αβ T cells, expanded from preexisting donor resident memory T cells (TRM) after encountering rapidly-repopulating recipient antigen-presenting cells in the graft mucosa, acquire effector T cell (Teff) functions and migrate into circulation and bone marrow (BM), where they attenuate HvG responses, facilitate donor cell engraftment and control rejection. Using single cell RNA-seq, we found that BM-infiltrating donor γδ T cells, dominated by “public” Vδ2 clonotypes, showed cytotoxic Teff phenotypes similar to CD8 αβ T cells, suggesting their potential to also attenuate HvG reactions and make space for donor hematopoietic stem and progenitor cell engraftment. We found that recipient γδ T cells gradually populated the allograft, undergoing phenotypic changes from Teff to TRM, mainly with “private” Vδ1 clonotypes. We hypothesize that γδ T cells not only participate in host defense against pathogens, but also have the potential to modulate two-way alloresponses after human ITx. We now propose a study of phenotypic and clonal tracking of human γδ T cells locally and systemically after ITx and further investigation of their regulatory roles on alloreactivity and association with graft outcomes. We propose to pursue three Specific Aims: To determine the chimerism, phenotype and clonotype of donor- (Aim 1) and recipient- (Aim 2) derived γδ T cells in graft mucosa, circulation and BM by flow cytometry, mass cytometry and TCRγδ-seq. Bulk TCRγδ-seq of pre-Tx unstimulated and ex vivo-stimulated γδ T cell repertoires and post-Tx γδ T cell repertoires in the circulation, intestinal allograft and BM of ITx patients will be linked with single cell transcriptional profiles of γδ T cell using the iRepertoire and 10x Genomics platforms (Aim 3). Our proposed research will provide a deeper understanding of the mechanisms behind γδ T cell chimerism, maturation of TRM features, and their modulatory roles on local and systemic alloresponses, facilitating the development of novel strategies to regulate γδ T cells to overcome rejection, infection and increase the utilization of ITx as a life-saving, quality of life-improving procedure.

项目总结