Analysis of Immune Escape from NK cell in Melanoma

黑色素瘤 NK 细胞的免疫逃逸分析

基本信息

批准号：
14570812
负责人：
KAGESHITA Toshiro
金额：
$ 2.18万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570812/
关键词：
melanoma NK cell immune escape MICA MICB

项目摘要

One of the larger concernsin melanoma T cell based immunotherapy is the immune escape phenomenon resulting from loss of melanoma associated antigens and/or HLA class I. The head investigator has been analyzing the mechanisms of down regulation of HLA class I in melanocytic cells.HLA class I was down regulated in melanoma lesions, which results in no T cell infiltration in such lesions. Theoretically loss of HLA class I in melanoma cells allows NK cell infiltration. However, most of the melanoma lesions show neither T cell nor NK cell infiltration. These data suggest that melanoma cells can escape from NK cell attack.In the present study, analysis of immune escape from NK cell in melanoma was investigated using monoclonal antibodies to MHC class I chain-related molecule A and B (MICA and MICB), which are ligands for the newly cloned NK cell activating receptor NKG2D.The head investigator found the following points :1)Expression of MICA in culture human melanoma cell linesTen of 16 human … More melanoma cell lines were shown to be MICA positive by ELISA.2)Expression of MICA in melanocytic tumorsExpression of MICA in melanocytic lesions was investigated immunohistochemically using frozen melanocytic nevus, and primary and metastatic melanoma lesions. MICA was not expressed in melanocytic nevus, however it was expressed in melanoma lesions. MICA was expressed in 30%, 60%, and 20% of radial growth phase, vertical growth phase, and metastatic lesions, respectively.3)Selection of monoclonal antibody to MICB which can react with formalin-fixed tissue sectionThe monoclonal antibody to MICA did not react with formalin-fixed tissue sections. Therefore, I tried to find a monoclonal antibody to MICB which can react with formalin-fixed tissue section from more than 10 antibodies to MICB. We were able to select one antibody to MICB that could react with formalin-fixed tissue sections using the retrieval antigen method.These data will facilitate the analysis of immune escape from NK cell in melanoma and further research project is under going at my laboratory. Less

基于黑色素瘤 T 细胞的免疫治疗中较大的问题之一是由于黑色素瘤相关抗原和/或 HLA I 类丢失而导致的免疫逃逸现象。首席研究员一直在分析黑素细胞中 HLA I 类下调的机制。 I 在黑色素瘤病变中表达下调，导致此类病变中没有 T 细胞浸润。理论上，黑色素瘤细胞中 HLA I 类的缺失允许 NK 细胞浸润。黑色素瘤病灶既没有 T 细胞也没有 NK 细胞浸润，这些数据表明黑色素瘤细胞可以逃避 NK 细胞的攻击。在本研究中，使用针对 MHC I 类链的单克隆抗体对黑色素瘤中 NK 细胞的免疫逃避进行了分析。相关分子A和B（MICA和MICB），它们是新克隆的NK细胞激活受体NKG2D的配体。首席研究员发现以下几点：1）MICA在培养人黑色素瘤细胞系 16 个人类黑色素瘤细胞系中的 10 个通过 ELISA 显示为 MICA 阳性。2）黑色素细胞肿瘤中 MICA 的表达使用冰冻黑色素细胞痣以及原发性和转移性黑色素瘤病变，通过免疫组织化学方法研究了黑色素细胞病变中 MICA 的表达。 MICA 在黑色素细胞痣中不表达，但在黑色素瘤病变中表达。径向生长期、垂直生长期、转移病灶分别为30%、60%、20%。 3)选择能与福尔马林固定组织切片反应的MICB单克隆抗体MICA单克隆抗体不与福尔马林反应因此，我试图从超过 10 种 MICB 抗体中找到一种能够与福尔马林固定组织切片发生反应的 MICB 单克隆抗体。使用修复抗原方法选择一种可以与福尔马林固定的组织切片发生反应的 MICB 抗体。这些数据将有助于分析黑色素瘤中 NK 细胞的免疫逃逸，并且我的实验室正在进行进一步的研究项目。