In depth characterization of Mtb specific HLA-E restricted human CD8+ T-cells

Mtb 特异性 HLA-E 限制性人类 CD8 T 细胞的深入表征

• 批准号: 9204701
• 负责人: Tom Ottenhoff
• 金额: $ 15.1万
• 依托单位: LEIDEN UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-28 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9204701
• 关键词: Aerosols; Alleles; Amino Acids; Animals; Antigen Presentation; B-Lymphocytes; Bacteria; Binding; Biological Markers; Blood; Blood Circulation; CD8B1 gene; Cells; Cessation of life; Characteristics; Chemicals; Clinical; Code; Collection; Data; Data Analyses; Diagnosis; Disease; Down-Regulation; Flow Cytometry; Frequencies; Funding; Future; Growth; HIV; HIV Infections; Human; Human Volunteers; IL4 gene; Immune system; Immunity; Individual; Infection; Interleukin-13; Interleukin-4; Interleukin-5; Investigation; Italy; Kinetics; Literature; Lung; Mediating; Molecular; Molecular Structure; Molecular Target; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; Outcome; Pathway interactions; Patients; Peptides; Peripheral Blood Mononuclear Cell; Phenotype; Population; Property; Proteins; RNA Interference; Reporting; Resistance; Rome; Route; Sampling; Signal Pathway; Sorting - Cell Movement; Staining method; Stains; T cell response; T-Cell Receptor; T-Lymphocyte; TNFSF10 gene; Tuberculosis; Tuberculosis Vaccines; Universities; Vaccinated; Vaccination; Vaccines; Validation; Variant; Work; aerosolized; clinically relevant; co-infection; cohort; combat; cytokine; cytotoxicity; follow-up; inhibitor/antagonist; insight; interest; nonhuman primate; novel; response; transcriptome sequencing; vaccination against tuberculosis; volunteer

1 Immunity to Mycobacterium tuberculosis (Mtb) has been the subject of detailed investigations for a number of 2 decades. Most studies have focused on classically restricted (MHC class Ia or II) T-cells, but recently, non- 3 classically restricted T-cells have become subject of increasing interest. Non-classical presentation molecules 4 (which include human CD1a,b,c,d; MR1; HLA-E and other HLA class Ib molecules) have been shown to 5 present Mtb antigens to human T-cells. HLA-E is a particularly interesting molecule, for a number of reasons: 6 first, there are only 2 coding variants described, which differ in only a single amino acid, located outside the 7 peptide binding groove, such that HLA-E essentially is a conserved, virtually monomorphic presentation 8 molecule; secondly, in contrast to HLA class Ia molecules, HLA-E is relatively resistant to HIV mediated 9 downregulation, such that antigen presentation should be maintained during HIV infection; thirdly, HLA-E is 10 present in Mtb containing intracellular compartments. Finally, we have recently shown that human CD8+ T- 11 cells recognizing Mtb peptides presented by HLA-E are present in the blood of Mtb infected individuals. 12 These cells have an unconventional phenotype and function: they had cytolytic properties, were able to 13 control intracellular outgrowth of Mtb, but unexpectedly produced mainly Th2 cytokines, including IL-4, IL-5 14 and IL-13, and efficiently helped B-cells through IL-4. 15 In the present project we propose an in depth characterization of these HLA-E restricted T-cells, both by 16 detailed characterization of available T-cell clones we have generated as well as by enumeration and 17 characterization of HLA-E restricted T-cells in (blood and BAL of) clinically relevant cohorts. The first part of 18 the project will characterize Mtb specific, HLA-E restricted T-cell clones, focusing first on their T-cell receptor 19 (TCR), including the requirements for peptide recognition and molecular structure of the TCR. In addition, the 20 clones will be employed to decipher the mechanism of intracellular Mtb growth inhibition, as this will be 21 important for understanding protective immunity towards Mtb. In the second part of the project, we will 22 enumerate HLA-E restricted T-cells in cohorts of Mtb infected individuals and individuals vaccinated with 23 BCG, both parenteral and aerosolized. These human studies will be bridged to parallel BCG vaccination 24 studies in non-human-primates, to allow validation of HLA-E restricted T-cell responses following vaccination 25 as well as following TB challenge. For all these studies unique collections of banked samples will be made 26 available by top edge collaborators (from UK, Italy and SA). Finally, tetramer (TM) positive cells will be sorted 27 to allow for RNA sequencing for full characterization of HLA-E restricted CD8+ T-cell responses. 28 All together these studies are expected to yield important new insights in the functional characteristics, clinical 29 relevance and vaccine kinetics of human Mtb specific, HLA-E restricted CD8+ T-cells. This will provide novel 30 insights into their potency as TB biomarkers and future TB vaccine targets.

1对结核分枝杆菌（MTB）的免疫力已成为许多详细研究的主题 20年。大多数研究都集中在经典限制（MHC类或II类）T细胞上，但最近，非 - 3经典限制的T细胞已成为越来越多的利益的主题。非古典表现分子 4（包括人CD1A，B，C，D; MR1； HLA-E和其他HLA类IB分子）已显示为 5呈现人类T细胞的MTB抗原。 HLA-E是一个特别有趣的分子，出于多种原因： 6首先，仅描述了2个编码变体，它们仅在一个位于外部的一个氨基酸中不同 7肽结合凹槽，因此HLA-E本质上是保守的，几乎是单态表现 8分子；其次，与HLA类IA分子相比，HLA-E对HIV介导的相对抗性 9下调，以便在HIV感染期间应保持抗原表现；第三，HLA-E是 10在含有细胞内室的MTB中存在。最后，我们最近表明人CD8+ T- 11个识别由HLA-E提供的MTB肽的细胞存在于MTB感染个体的血液中。 12这些细胞具有非常规的表型和功能：它们具有细胞溶解特性，能够能够 13控制MTB的细胞内生长，但出乎意料地产生的主要是Th2细胞因子，包括IL-4，IL-5 14和IL-13，并有效地通过IL-4帮助B细胞。 15在本项目中，我们提出了这些HLA-E限制的T细胞的深入表征 16我们已经生成的可用T细胞克隆的详细表征以及枚举以及 17临床相关队列中（血液和BAL）中HLA-E受限的T细胞的表征。第一部分 18该项目将以MTB为特定的HLA-E受限的T细胞克隆，首先关注其T细胞受体 19（TCR），包括对TCR的肽识别和分子结构的要求。另外， 将使用20个克隆来破译细胞内MTB生长抑制的机制，因为这将是 21对了解MTB的保护性免疫很重要。在项目的第二部分中，我们将 22列举MTB感染的个体和接种疫苗的人群中的HLA-E限制的T细胞 23 BCG，均和气溶解。这些人类研究将被桥接到平行的BCG疫苗接种 24个非人类拟南芥的研究，以验证疫苗接种后HLA-E限制的T细胞反应 25以及遵循结核病挑战。对于所有这些研究 26由Top Edge合作者（来自英国，意大利和SA）获得。最后，将对四聚体（TM）阳性细胞进行排序 27允许进行RNA测序，以完全表征HLA-E受限的CD8+ T细胞响应。 28所有这些研究预计将在功能特征，临床方面产生重要的新见解 29人类MTB特异性HLA-E受限的CD8+ T细胞的相关性和疫苗动力学。这将提供小说 30个洞察力作为结核病生物标志物和未来结核病疫苗靶标的效力。