Functional genetics of human innate immunity in the bimodal gamma delta T cell response to Epstein-Barr Virus and in education of NK cells and their re-education to respond to autologous cells

人类先天免疫的功能遗传学在双峰 γδT 细胞对 Epstein-Barr 病毒的反应以及 NK 细胞的教育及其对自体细胞作出反应的再教育中

• 批准号: 10552637
• 负责人: PETER R PARHAM
• 金额: $ 39.13万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-05 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10552637
• 关键词: Activated Natural Killer Cell; Address; Adolescent; Adult; Affect; Allogenic; Autologous; Blood Cells; Burkitt Lymphoma; Cancer Etiology; Cancer Patient; Cell Line; Cell physiology; Cells; Child; Clinical; Coculture Techniques; Communicable Diseases; Complement; Complex; Costs and Benefits; Cytomegalovirus; Data; Development; Diagnosis; Disease; Donor person; Education; Educational Status; Epitopes; Epstein-Barr Virus Infections; Epstein-Barr Virus latency; Equilibrium; Exhibits; Exposure to; Foundations; Frequencies; Gender; Gene Family; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Graft-Versus-Tumor Induction; HLA Class I Genes; HLA-A gene; Haplotypes; High-Throughput RNA Sequencing; Human; Human Genetics; Human Herpesvirus 4; Immune; Immune response; Immunogenetics; Immunoglobulins; Immunotherapy; In Vitro; Individual; Infection; Infectious Mononucleosis; Innate Immune Response; Innate Immune System; Investigation; KLRD1 gene; Killer Cells; Kinetics; Leukocytes; Ligands; Lymphocyte; Lymphoma cell; Lytic Phase; MHC Class I Genes; Malignant Neoplasms; Mediating; Modality; Modeling; Molecular; Natural Immunity; Natural Killer Cell Immunotherapy; Natural Killer Cells; Nature; Patients; Peptides; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Physiological; Population; Predisposition; Proliferating; Receptor Gene; Regulation; Research; Residual Neoplasm; Resolution; Rest; Signal Transduction; Study models; Surface; System; T cell response; T-Lymphocyte; T-cell receptor repertoire; Variant; Work; adaptive immunity; cancer cell; cell killing; cohort; experimental study; hematopoietic cell transplantation; improved; in vitro Model; in vivo; insight; latent infection; leukemia; neoplastic cell; peripheral blood; receptor; receptor expression; responders and non-responders; response; segregation; sex; transcriptome; transcriptome sequencing; treatment choice; tumor; γδ T cells

This project investigates how human genetic variation diversifies innate immune responses to infection and cancer. Aim 1 concerns innate immunity to Epstein-Barr Virus (EBV), a cause of cancer. We found humans form two equally sized groups based on their response to EBV. Group 1 make strong Natural Killer (NK) cell and Vγ9Vδ2 T cell responses, whereas Group 2 combine a vigorous NK cell response with a poor Vγ9Vδ2 T cell response. This difference does not correlate with sex, HLA type, prior exposure to EBV or CMV, abundance of Vγ9Vδ2 T cells, or their functionality in the two groups. We propose that this even balance of responders and non-responders arises because of costs and benefits in making a Vγ9Vδ2 T cell response to EBV. Genetic precedents from our previous work are functionally distinctive haplotypes of both the killer cell immunoglobulin-like receptor (KIR) gene family, and the HLA class I gene family. We will determine genetic and functional differences that distinguish Group 1 and 2 donors, with three approaches. First, we will define the γδ T cell receptor repertoires for resting and EBV-activated γδ T cells of Group 1 and 2 donors. Second, we will define the phenotype and function of EBV-responding Vγ9Vδ2 T cells and NK cells in Group 1 and 2 individuals. Third, the γδ T cells of Group 1 and 2 individuals will be compared by high throughput RNA sequencing. Aim 2 examines NK cell and γδ T cell crosstalk in response to EBV. Our preliminary data shows that γδ T cells have an inhibitory effect on NK cell function in the context of EBV, and that this effect is distinct in Group 1 vs Group 2 donors. We propose an examination of NK cell and γδ T cell crosstalk that incorporates both NK cell educational status, and the dichotomous γδ T cell responses to EBV. Aim 3 examines interactions between KIR and HLA-A, B and C that control NK cell development and responses to infected or malignant cells that have low expression of HLA class I. These interactions involve four HLA class I epitopes and four inhibitory KIR (iKIR). The polymorphism of KIR and HLA, their independent segregation and stochastic KIR expression, mean that most humans express ligands without receptors and vice versa. During development, NK cells expressing an HLA epitope and its cognate iKIR, become educated to kill cells that lack the epitope. Consequently, educated NK cells can instigate a graft-versus-tumor (GVT) response in leukemia patients following allogeneic hematopoietic cell transplant. The GVT response clears residual tumor cells and improves patient survival. Preliminary experiments demonstrate that NK cells can be re-educated in vitro to kill malignant cells. We will determine the potential for re-education in a donor cohort representing all 12 common genetic combinations of the 4 HLA epitopes. This will give new insight on NK cell education and provide the foundation for re-educating autologous NK cells as immunotherapy for leukemia. In summary, this work will define the genetic and mechanistic parameters of three phenomena: the dichotomous Vγ9Vδ2 T cell response to EBV, Vγ9Vδ2T cell crosstalk with NK cells, and in vitro NK cell re-education.

该项目研究人类遗传变异如何使对感染和感染的先天免疫反应多样化 癌症。目标 1 涉及对 Epstein-Barr 病毒 (EBV) 的先天免疫，EBV 是癌症的一种原因。我们发现了人类 根据他们对 EBV 的反应分成两个同等规模的小组。第 1 组产生强大的自然杀伤 (NK) 细胞 和 Vγ9Vδ2 T 细胞反应，而第 2 组结合了强烈的 NK 细胞反应和较差的 Vγ9Vδ2 T 细胞反应 细胞反应。这种差异与性别、HLA 类型、之前接触过 EBV 或 CMV 无关， Vγ9Vδ2 T 细胞的丰度，或其在两组中的功能。我们建议，这种均匀平衡 反应者和非反应者的出现是因为 Vγ9Vδ2 T 细胞对 EB病毒。我们之前工作的遗传先例是杀伤细胞的功能独特的单倍型 免疫球蛋白样受体 (KIR) 基因家族和 HLA I 类基因家族。我们将确定遗传 以及区分第 1 组和第 2 组捐赠者的功能差异，采用三种方法。首先，我们将定义 第 1 组和第 2 组供体的静息和 EBV 激活的 γδ T 细胞的 γδ T 细胞受体库。第二，我们 将定义第 1 组和第 2 组中 EBV 反应性 Vγ9Vδ2 T 细胞和 NK 细胞的表型和功能 个人。第三，将通过高通量RNA比较第1组和第2组个体的γδ T细胞 测序。目标 2 检查 NK 细胞和 γδ T 细胞对 EBV 的反应。我们的初步数据显示 γδ T 细胞在 EBV 背景下对 NK 细胞功能具有抑制作用，并且这种作用是明显的 在第 1 组与第 2 组捐赠者中。我们建议对 NK 细胞和 γδ T 细胞串扰进行检查，其中包括 NK 细胞的教育状况以及对 EBV 的二分 γδ T 细胞反应。目标 3 检查相互作用 KIR 与 HLA-A、B 和 C 之间，控制 NK 细胞发育以及对感染或恶性细胞的反应 HLA I 类低表达的细胞。这些相互作用涉及四个 HLA I 类表位和四个 抑制性 KIR (iKIR)。 KIR和HLA的多态性、独立分离和随机KIR 表达，意味着大多数人表达配体而不表达受体，反之亦然。在开发过程中， 表达 HLA 表位及其同源 iKIR 的 NK 细胞被训练去杀死缺乏该表位的细胞。 因此，受过训练的 NK 细胞可以激发白血病患者的移植物抗肿瘤 (GVT) 反应 同种异体造血细胞移植后。 GVT 反应可清除残留肿瘤细胞并改善 患者生存。初步实验表明NK细胞可以在体外进行再教育以杀死恶性细胞 细胞。我们将确定代表所有 12 种常见遗传基因的捐赠者群体接受再教育的可能性 4 个 HLA 表位的组合。这将为 NK 细胞教育提供新的见解并提供基础 用于重新培养自体 NK 细胞作为白血病的免疫疗法。总之，这项工作将定义 三种现象的遗传和机械参数：对 EBV 的二分 Vγ9Vδ2 T 细胞反应， Vγ9Vδ2T 细胞与 NK 细胞的串扰，以及体外 NK 细胞的再教育。

项目成果

Natural LILRB1 D1-D2 variants show frequency differences in populations and bind to HLA class I with various avidities.

• DOI: 10.1007/s00251-022-01264-7
• 发表时间: 2022-12
• 期刊: IMMUNOGENETICS
• 影响因子: 3.2
• 作者: Liu, Fuguo;Cocker, Alexander T. H.;Pugh, Jason L.;Djaoud, Zakia;Parham, Peter;Guethlein, Lisbeth A.
• 通讯作者: Guethlein, Lisbeth A.

Abundant CpG-sequences in human genomes inhibit KIR3DL2-expressing NK cells.

• DOI: 10.7717/peerj.12258
• 发表时间: 2021
• 期刊: PeerJ
• 影响因子: 2.7
• 作者: Pugh J;Guethlein L;Parham P
• 通讯作者: Parham P

Phosphoantigen-Stimulated γδ T Cells Suppress Natural Killer-Cell Responses to Missing-Self.

• DOI: 10.1158/2326-6066.cir-21-0696
• 发表时间: 2022-05-03
• 期刊: CANCER IMMUNOLOGY RESEARCH
• 影响因子: 10.1
• 作者: Walwyn-Brown, Katherine;Pugh, Jason;Cocker, Alexander T. H.;Beyzaie, Niassan;Singer, Bernhard B.;Olive, Daniel;Guethlein, Lisbeth A.;Parham, Peter;Djaoud, Zakia
• 通讯作者: Djaoud, Zakia

CD56-negative NK cells: Frequency in peripheral blood, expansion during HIV-1 infection, functional capacity, and KIR expression.

• DOI: 10.3389/fimmu.2022.992723
• 发表时间: 2022
• 期刊: FRONTIERS IN IMMUNOLOGY
• 影响因子: 7.3
• 作者: Cocker, Alexander T. H.;Liu, Fuguo;Djaoud, Zakia;Guethlein, Lisbeth A. A.;Parham, Peter
• 通讯作者: Parham, Peter