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.

该项目调查了人类遗传变异如何使先天免疫回报与感染和 癌症。 AIM 1涉及对Epstein-Barr病毒（EBV）的先天免疫力，这是癌症的原因。我们找到了人类 根据两个同等大小的组对EBV的反应。第1组制造强大的天然杀手（NK）细胞 和Vγ9Vδ2T细胞反应，而组2将剧烈的NK细胞响应与较差的Vγ9Vδ2T结合 细胞反应。这种差异与性别，HLA类型，事先暴露于EBV或CMV无关， Vγ9VΔ2T细胞的抽象或两组中其功能。我们建议这种平衡 响应者和非反应者由于成本和收益而产生 EBV。我们先前工作的遗传先例是两个杀手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。我们将确定通用 以及以三种方法区分第1和2组捐助者的功能差异。首先，我们将定义 第1组和2个供体的γδT细胞受体曲目用于静止和EBV激活的γδT细胞。第二，我们 将定义第1和2组中EBV反应的Vγ9Vδ2T细胞和NK细胞的表型和功能 个人。第三，第1组和2个个体的γδT细胞将通过高吞吐量RNA进行比较 测序。 AIM 2检查响应EBV的NK细胞和γδT细胞串扰。我们的初步数据显示 在EBV的背景下，γδT细胞对NK细胞功能具有抑制作用，并且这种作用是不同的 在第1组与第2组捐助者中。我们提出了对NK细胞和γδT细胞串扰的检查 NK细胞教育状态和二分法γδT细胞对EBV的反应。 AIM 3检查互动 在控制NK细胞发育的KIR和HLA-A，B和C之间，对感染或恶性的反应 HLA类I表表达低的细胞I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，它们的独立分离和随机KIR 表达，意味着大多数人表达没有受体的配体，反之亦然。在开发期间， 表达HLA表位及其同源ikir的NK细胞受到教育，以杀死缺乏表位的细胞。 因此，受过教育的NK细胞可以激发白血病患者的移植物肿瘤（GVT）反应 遵循同种异体造血细胞移植。 GVT反应清除残留肿瘤细胞并改善 患者生存。初步实验表明，可以在体外重新教育NK细胞以杀死恶性肿瘤 细胞。我们将确定代表所有12个常见遗传的捐赠者队列中重新教育的潜力 4 HLA表位的组合。这将为NK细胞教育提供新的见解，并为基础提供基础 用于重新教育自体NK细胞作为白血病的免疫疗法。总而言之，这项工作将定义 三种现象的遗传和机械参数：二分法Vγ9Vδ2T细胞对EBV的反应， Vγ9Vδ2T细胞与NK细胞串扰，并在体外NK细胞重新教育。

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