Formation of human NK cell repertoires: role of HLA class I and KIR gene polymorphism

人类 NK 细胞库的形成：HLA I 类和 KIR 基因多态性的作用

基本信息

批准号：
228837322
负责人：
Professor Dr. Markus G. Uhrberg
金额：
--
依托单位：
Klinik für Kinder-Onkologie, -Hämatologie und Klinische Immunologie
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2013
资助国家：
德国
起止时间：
2012-12-31 至 2017-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/228837322?language=en
关键词：
Formation human NK cell repertoires

项目摘要

Natural killer (NK) cells serve an important function in the early control of virus infection and the eradication of malignant cells in the course of tumor surveillance. In humans, HLA class I-specific NK receptors of the KIR and NKG2 gene families play a crucial role in this process. These receptors are expressed on NK cells in different combinations and determine the specificity against HLA class I-deficient target cells. It is currently unclear if the formation of NK cell repertoires is primarily exogenously influenced by the stem cell niche and the local expression of HLA ligands or endogenously by genetic factors such as the polymorphic KIR genes. In the present project these questions will be addressed in several in vitro models of NK cell differentiation. To this end, early hematopoietic progenitors will be co-cultured with HLA class I-transfected murine stroma cells and differentiated to mature NK cells. Alternatively mesenchymal stem cells (MSC) will be employed as accessory cells enabling to follow the NK cell differentiation process in a human stem cell niche. To dissect the process of receptor acquisition more closely, NKG2A and selected KIR genes will be knocked down by RNAi as well as stably overexpressed at the hematopoietic progenitor stage. As an alternative stem cell source, we will employ induced pluripotent stem (iPS) cells for NK cell differentiation. The formation of NK repertoires and the acquisition of effector function will be analyzed on a clonal basis by multicolor flow cytometry analysis, using a panel of NKG2A and KIR-specific antibodies as well as functional markers. These experiments should help to understand in how far the program of NK repertoire formation is hard-wired or if rather KIR ligands and the stem cell niche as major exogenous determinants influence this process. Importantly, the results could serve as a model to better understand NK cell reconstitution following hematopoietic stem cell transplantation and might pave the way for future immunotherapeutic applications employing specific in vitro generated and expanded NK cells.

天然杀伤（NK）细胞在肿瘤监测过程中早期控制病毒感染和消除恶性细胞的早期控制中起着重要功能。在人类中，KIR和NKG2基因家族的HLA I类特异性NK受体在此过程中起着至关重要的作用。这些受体以不同的组合在NK细胞上表达，并确定针对HLA I类缺乏靶细胞的特异性。目前尚不清楚NK细胞库的形成是主要受干细胞生态位和HLA配体的局部表达的外源影响还是由遗传因子（例如多态KIR基因）进行内源性。在本项目中，这些问题将在几种NK细胞分化的体外模型中解决。为此，早期造血祖细胞将与HLA I类转染的鼠基质细胞共同培养，并与成熟的NK细胞区分开。替代性间充质干细胞（MSC）将被用作辅助细胞，使得能够跟随人类干细胞生态位的NK细胞分化过程。为了更紧密地剖析受体获取的过程，RNAi将击倒NKG2A和选定的KIR基因，并在造血祖细胞阶段稳定过表达。作为另一种干细胞来源，我们将利用诱导的多能干（IPS）细胞进行NK细胞分化。使用NKG2A和KIR特异性抗体以及功能标记物，将通过多色流式细胞仪分析和KIR特异性抗体以及功能标记来分析NK曲目的形成和效应功能的获取。这些实验应该有助于了解NK曲目形成的程序是硬线的多远，或者是KIR配体和作为主要外源决定因素会影响这一过程的kir配体和干细胞生态位。重要的是，该结果可以作为造血干细胞移植后更好地理解NK细胞重建的模型，并可能为未来的免疫治疗应用铺平道路，该应用采用特定的体外产生和扩展的NK细胞。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

NK cell development in a human stem cell niche: KIR expression occurs independently of the presence of HLA class I ligands.

人类干细胞生态位中的 NK 细胞发育：KIR 表达的发生独立于 HLA I 类配体的存在

DOI：
10.1182/bloodadvances.2018019059
发表时间：
2018
期刊：
Blood advances
影响因子：
7.5
作者：
Xiaoyi Zhao;Sandra Weinhold;Jens Brands;Maryam Hejazi;Özer Degistirici;Gesine Kögler;Roland Meisel;Markus Uhrberg
通讯作者：
Markus Uhrberg

Human KIR repertoires: shaped by genetic diversity and evolution