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: 10493378
• 负责人: Jianing Fu
• 金额: $ 20.25万
• 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 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10493378
• 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 γδT细胞可以识别不同的抗原并发挥不同的功能。与生俱来的和适应性的特征 的人类γδT细胞可能是由不同的γδT细胞受体(TcR)谱系驱动的，它可以通过以下方式形成 组织分区、年龄和抗原暴露史。尽管占了相当大的比例 包括肠道和肝脏在内的许多器官中驻留的T细胞、γδT细胞及其在移植中的可能作用 结果在很大程度上没有得到充分的研究。我们的首要目标是阐明 γδT细胞参与同种异体肠道移植排斥反应和宿主防御的机制 移植(ITX)。我们发现，ITX后的临床结果在很大程度上取决于平衡。 移植物抗宿主(GVH)和宿主抗移植物(HVG)同种异体反应之间的关系，使用一种识别 TcR、β、CDR3高表达的TX前混合淋巴细胞反应后的同种异体反应性T细胞克隆 吞吐量测序(TCRβ-seq)。我们发现，GVH反应性αβT细胞从先前存在的 供体驻留记忆T细胞(TRM)在遇到快速繁殖的受体抗原提呈细胞后 在移植物粘膜中，获得效应T细胞(Tef)功能，并迁移到循环和骨髓(BM)中， 在那里它们可以减弱HVG反应，促进供体细胞植入和控制排斥反应。使用单个单元格 Rna-seq，我们发现BM渗入的供者γδT细胞，主要是公共的Vδ2克隆型，表现出 与CD8αβT细胞相似的细胞毒性T细胞表型，表明它们也有可能减弱hvg反应 并为供者造血干细胞和祖细胞植入腾出空间。我们发现收件人γδT 同种异体移植物的细胞逐渐充填，经历了从Tef到Trm的表型变化，主要是“私人”。 Vδ1克隆型。我们假设γδT细胞不仅参与宿主对病原体的防御，而且 也有可能调节人类ITX后的双向同种异体反应。我们现在提议研究一下 ITX后人γδT细胞的局部和系统表型及克隆性示踪及进一步研究 它们对同种异体反应性的调节作用以及与移植结果的关联。我们建议追求三个目标 具体目标：确定供体-(目标1)和受体-(目标2)的嵌合体、表型和克隆 用流式细胞仪、质量细胞仪和TCRγδ-SEQ检测移植物黏膜、循环和骨髓中的CD8+T细胞。散装 TX前未刺激和体外刺激的γδT细胞谱系和TX后γδT细胞的TCRγδ-SEQ ITX患者的循环、同种异体肠移植和骨髓中的谱系将与单细胞相连 使用iRepertoire和10x基因组学平台的γδT细胞转录图谱(目标3)。我们的建议 研究将提供更深入的了解γδT细胞嵌合、成熟背后的机制 TRM特征及其对局部和全身同种异体反应的调节作用，促进 调节γδT细胞的新策略以克服排斥、感染和提高其作为一种治疗手段的利用率 挽救生命，提高生活质量的程序。