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

该项目研究人类遗传变异如何使对感染的先天免疫反应多样化， 癌目的1涉及对EB病毒（EBV）的先天免疫，EB病毒是癌症的一种原因。我们发现了人类 根据他们对EBV的反应分成两个同等大小的组。第1组产生强大的自然杀伤（NK）细胞 和Vγ 9 V δ2 T细胞应答，而组2联合收割机结合了强有力的NK细胞应答和弱的Vγ 9 V δ2 T细胞应答。 细胞反应。这种差异与性别、HLA类型、既往暴露于EBV或CMV无关， 两组中Vγ 9VS 2 T细胞的丰度或其功能性。我们建议， 应答者和无应答者的出现是因为使Vγ 9VS 2 T细胞应答于 EB病毒从我们以前的工作遗传先例是功能独特的单倍型的杀手细胞和 免疫球蛋白样受体（KIR）基因家族和HLA I类基因家族。我们将确定基因 和区分第1组和第2组供体的功能差异，有三种方法。首先，我们将定义 第1组和第2组供体的静息和EBV活化的γ δ T细胞的γδ T细胞受体库。二是 将定义组1和组2中EBV应答性Vγ 9VS 2 T细胞和NK细胞的表型和功能 个体第三，将通过高通量RNA比较组1和组2个体的γδ T细胞 测序目的2检测NK细胞和γδ T细胞对EBV应答的相互作用。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，并且这种作用是独特的， 第1组与第2组供体。我们提出了一个检查NK细胞和γδ T细胞的串扰， NK细胞教育状况和对EBV的二分γδ T细胞应答。目标3检查相互作用 KIR与HLA-A、B和C之间的关系，这些基因控制NK细胞发育和对感染或恶性肿瘤的应答。 低表达HLA I类的细胞。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的多态性及其独立分离和随机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细胞被训练以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 异基因造血细胞移植后死亡GVT反应清除残留的肿瘤细胞，并改善 患者生存率。初步实验表明，NK细胞可以在体外再训练，以杀死恶性肿瘤。 细胞我们将在代表所有12种常见遗传病的供体队列中确定再教育的可能性。 4种HLA表位的组合。这将为NK细胞教育提供新的见解， 用于再训练自体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制参数：二分Vγ9Vδ2 T细胞对EBV的应答， Vγ 9VS 2 T细胞与NK细胞的串扰，以及体外NK细胞再教育。

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.