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

• 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

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.

自然杀伤（NK）细胞在肿瘤监测过程中早期控制病毒感染和消灭恶性细胞方面发挥着重要作用。在人类中，KIR 和 NKG2 基因家族的 HLA I 类特异性 NK 受体在此过程中发挥着至关重要的作用。这些受体以不同的组合在 NK 细胞上表达，并决定针对 HLA I 类缺陷靶细胞的特异性。目前尚不清楚 NK 细胞库的形成主要受干细胞生态位和 HLA 配体局部表达的外源影响，还是受多态性 KIR 基因等遗传因素的内源影响。在本项目中，这些问题将在几个 NK 细胞分化的体外模型中得到解决。为此，早期造血祖细胞将与 HLA I 类转染的小鼠基质细胞共培养，并分化为成熟的 NK 细胞。或者，间充质干细胞 (MSC) 将被用作辅助细胞，能够跟踪人类干细胞生态位中的 NK 细胞分化过程。为了更仔细地剖析受体获得的过程，NKG2A 和选定的 KIR 基因将被 RNAi 敲低，并在造血祖细胞阶段稳定过度表达。 作为替代干细胞来源，我们将采用诱导多能干 (iPS) 细胞进行 NK 细胞分化。将使用一组 NKG2A 和 KIR 特异性抗体以及功能标记，通过多色流式细胞术分析在克隆基础上分析 NK 库的形成和效应子功能的获得。这些实验应该有助于了解 NK 库形成程序的硬连线程度，或者更确切地说，KIR 配体和干细胞生态位作为主要外源决定因素影响这一过程。重要的是，这些结果可以作为更好地了解造血干细胞移植后 NK 细胞重建的模型，并可能为未来使用特定的体外生成和扩增的 NK 细胞进行免疫治疗应用铺平道路。

项目成果

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

Age-related changes in natural killer cell repertoires: impact on NK cell function and immune surveillance

• DOI: 10.1007/s00262-015-1750-0
• 发表时间: 2016-04-01
• 期刊: CANCER IMMUNOLOGY IMMUNOTHERAPY
• 影响因子: 5.8
• 作者: Manser, Angela R.;Uhrberg, Markus
• 通讯作者: Uhrberg, Markus

Impaired cytotoxicity associated with defective natural killer cell differentiation in myelodysplastic syndromes

• DOI: 10.3324/haematol.2014.118679
• 发表时间: 2015-05-01
• 期刊: HAEMATOLOGICA
• 影响因子: 10.1
• 作者: Hejazi, Maryam;Manser, Angela R.;Uhrberg, Markus
• 通讯作者: Uhrberg, Markus