Memory, Phenotype, and Function of TB-reactive Human MR1 Restricted T cells

结核病反应性人 MR1 限制性 T 细胞的记忆、表型和功能

• 批准号: 10329945
• 负责人: DAVID M. LEWINSOHN
• 金额: $ 56.32万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10329945
• 关键词: Activities of Daily Living; Address; Adjuvant; Adult; Aerosols; Anabolism; Anatomy; Antigens; Bacteria; Blood Circulation; CD8-Positive T-Lymphocytes; Cell surface; Cells; Child; Clone Cells; Communicable Diseases; Data Analyses; Dependence; Development; Diagnosis; Disease; Early Diagnosis; Environment; Environmental Exposure; Evaluation; Exposure to; Frequencies; Future; Granzyme; Growth; HIV; Human; Immune system; Immunity; Immunobiology; Immunocompetent; Immunologic Memory; Immunotherapy; Individual; Infant; Infection; Infection Control; Inflammatory; Ligands; Location; Lung; Lung Capacity; Major Histocompatibility Complex; Memory; Microbe; Mononuclear; Mucous Membrane; Mus; Mycobacterium Infections; Mycobacterium tuberculosis; Organism; Outcome; Pathway interactions; Peripheral Blood Mononuclear Cell; Phenotype; Play; Progressive Disease; Property; Proteins; Pulmonary Tuberculosis; Riboflavin; Role; T-Lymphocyte; T-Lymphocyte Subsets; T-cell receptor repertoire; Thymus Gland; Time; Tuberculosis; Tuberculosis Vaccines; Umbilical Cord Blood; Vaccination; Vaccines; antimicrobial; cohort; fungus; granulysin; improved; microbial; mortality; mucosal site; mycobacterial; novel; peripheral blood; response; transcriptome sequencing; treatment effect; vaccination strategy

Project Summary Mucosal associated invariant T (MAIT) cells, a subset of T cells restricted by MR1 (MR1T cells), are an innate- like T cell subset prevalent in humans and distributed throughout mucosal sites. Human MAIT cells are defined by the expression of the semi-invariant TCRα chain TRAV1-2/TRAJ12/20/33, restriction by the non- polymorphic major histocompatibility complex (MHC) class I-like molecule, MHC-related protein 1 (MR1), and by recognition of small organic molecules, derived from riboflavin biosynthesis. We have found that MAIT cells are both “innate” effectors as evidenced by their functionality in the thymus, and have the capacity to adapt to their environment as evidenced by their effector memory cell surface phenotype and selective TCR usage. However, it is not clear if these expansions are driven by ongoing microbial or environmental exposures, or if MAIT cells retain the capacity to respond selectively to future antigenic challenges. If MAIT cells have memory, then they could be harnessed in future vaccination strategies. We provide evidence demonstrating oligoclonal expansions of pro-inflammatory MAIT cells in the human airway of subjects infected with TB. We also provide evidence that vaccination with BCG can result in functional MAIT cell expansion, and provide evidence that MAIT cells have an anti-microbial, polycytotoxic phenotype. We postulate that MR1T cells can facilitate the control of infection with Mtb, and will explore whether or not these cells have immunologic memory. Specific Aim 1: To determine whether or not CD8+ MR1T cells have features of immunologic memory. By determining the TCR repertoire of MR1T cells prior to antigenic exposure (human cord blood mononuclear cells; CBMC), following antigenic exposure as a result of infant vaccination with BCG, in the setting of TB (adult PBMC), in the lungs (BAL) and peripheral blood of subjects with pulmonary TB at the time of diagnosis and following treatment, we will establish if MAIT cells demonstrate TCR usage that reflects antigenic exposure, persistence following antigenic exposure, and dependence on the presence of antigen. Specifically, we anticipate that we will observe a broad repertoire of MR1T cell TCRs in human cord blood, and that in adults we will observe a narrower repertoire characterized by oligocolonal expansions. Following vaccination with BCG and in TB, we would expect to observe oligoclonal expansions in the PBMC and BAL, and would predict that these cells will return to the circulation following treatment. Finally, we will ask if these TCRs are associated with discrete ligand recognition. Specific Aim 2: To define the phenotype and functional capacity of MR1T cells. A combination of flow cytometric analysis of both ex vivo MR1T cells and MR1T cell clones, expression data analysis of ex vivo MR1T cells, and mechanistic evaluation of MR1T cell clones derived from each of the cohorts and time points described will be used to define the full functional and anti-microbial phenotype of these cells.

项目摘要 粘液相关不变T（MAIT）细胞是受MR 1限制的T细胞亚群（MR 1 T细胞），是一种先天性免疫缺陷细胞。 如T细胞亚群普遍存在于人类并分布在整个粘膜部位。人MAIT细胞被定义为 通过表达半恒定TCRα链TRAV 1 -2/TRAJ 12/20/33，通过非- 多态性主要组织相容性复合体（MHC）I类分子，MHC相关蛋白1（MR 1），和 通过识别小的有机分子，衍生自核黄素的生物合成。我们发现MAIT细胞 都是“先天”效应器，如它们在胸腺中的功能所证明的，并且具有适应 它们的环境，如通过它们的效应记忆细胞表面表型和选择性TCR使用证明的。 然而，目前尚不清楚这些扩张是否是由持续的微生物或环境暴露驱动的，或者 MAIT细胞保留了对未来抗原挑战选择性反应的能力。如果MAIT细胞具有 记忆，那么它们就可以在未来的疫苗接种策略中得到利用。 我们提供的证据表明，促炎性MAIT细胞在人类中的寡克隆扩增， 肺结核患者的呼吸道。我们还提供了接种卡介苗可导致 功能性MAIT细胞扩增，并提供MAIT细胞具有抗微生物、多细胞毒性 表型我们假设MR 1 T细胞可以促进Mtb感染的控制，并将探索 这些细胞是否具有免疫记忆。 具体目的1：确定CD 8 + MR 1 T细胞是否具有免疫记忆特征。 通过在抗原暴露前测定MR 1 T细胞的TCR库（人脐带血单核细胞）， 在结核病背景下，由于婴儿接种卡介苗而导致抗原暴露后， 在诊断时肺TB受试者的肺（BAL）和外周血中， 在治疗后，我们将确定MAIT细胞是否表现出反映抗原性的TCR使用， 暴露、抗原暴露后的持久性和对抗原存在的依赖性。具体地说， 我们预期，我们将在人脐带血中观察到广泛的MR 1 T细胞TCR库， 成年人，我们将观察到以寡结肠扩张为特征的较窄的库。接种后 在BCG和TB中，我们预期在PBMC和BAL中观察到寡克隆扩增， 预测这些细胞将在治疗后返回循环。最后，我们会问这些TCR是否 与离散配体识别相关。 具体目的2：确定MR 1 T细胞的表型和功能能力。 结合离体MR 1 T细胞和MR 1 T细胞克隆的流式细胞术分析，表达数据 离体MR 1 T细胞的分析，以及来源于每一种细胞的MR 1 T细胞克隆的机制评估。 将使用所描述的队列和时间点来定义以下的全功能和抗微生物表型： 这些细胞。

项目成果

Production of Proinflammatory Cytokines by CD4+ and CD8+ T Cells in Response to Mycobacterial Antigens among Children and Adults with Tuberculosis.

• DOI: 10.3390/pathogens12111353
• 发表时间: 2023-11-14
• 期刊: Pathogens (Basel, Switzerland)
• 作者: Morrow E;Liu Q;Kiguli S;Swarbrick G;Nsereko M;Null MD;Cansler M;Mayanja-Kizza H;Boom WH;Chheng P;Nyendak MR;Lewinsohn DM;Lewinsohn DA;Lancioni CL
• 通讯作者: Lancioni CL

The Missing Link in Correlates of Protective Tuberculosis Immunity: Recognizing the Infected Cell.

• DOI: 10.3389/fimmu.2022.869057
• 发表时间: 2022
• 期刊: Frontiers in immunology
• 影响因子: 7.3

Optimising the yield from bronchoalveolar lavage on human participants in infectious disease immunology research.

• DOI: 10.1038/s41598-023-35723-2
• 发表时间: 2023-05-31
• 期刊: Scientific reports
• 影响因子: 4.6

Postnatal Expansion, Maturation, and Functionality of MR1T Cells in Humans.

• DOI: 10.3389/fimmu.2020.556695
• 发表时间: 2020
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Swarbrick GM;Gela A;Cansler ME;Null MD;Duncan RB;Nemes E;Shey M;Nsereko M;Mayanja-Kizza H;Kiguli S;Koh J;Hanekom WA;Hatherill M;Lancioni C;Lewinsohn DM;Scriba TJ;Lewinsohn DA
• 通讯作者: Lewinsohn DA

Functional and Activation Profiles of Mucosal-Associated Invariant T Cells in Patients With Tuberculosis and HIV in a High Endemic Setting.

• DOI: 10.3389/fimmu.2021.648216
• 发表时间: 2021
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Balfour A;Schutz C;Goliath R;Wilkinson KA;Sayed S;Sossen B;Kanyik JP;Ward A;Ndzhukule R;Gela A;Lewinsohn DM;Lewinsohn DA;Meintjes G;Shey M
• 通讯作者: Shey M