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Study of EBNA1-specific cellular immune responses in EB-virus associated diseases in chilhood

儿童 EB 病毒相关疾病中 EBNA1 特异性细胞免疫反应的研究

基本信息

批准号：
18591212
负责人：
ITO Yoshinori
金额：
$ 2.57万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591212/
关键词：
Epstein-Barr virus EBV-encoded nuclear antigen 1 cytotoxic T lymphocyte enitone MHC-tetramer

项目摘要

Epstein-Barr virus (EBV) -encoded nuclear antigen I (EBNA1) is an attractive target for immunotherapy against EBV-associated malignancies because it is expressed in all EBV-positive cells. Although CD^8+ cytotoxic T-lymphocyte (CTL) epitope presentation is largely prevented by its unique domain, the use of mRNA-transduced dendritic cells (DCs) would offer the advantage of priming EBNAl-specific CTLs. After stimulation with EBNA1 transduced monocyte-derived DCs, we successfully isolated two EBNA1-specific OTL clones B5 and C6 from a healthy donor. These Oils recognize peptides in the context of HLA-B*3501 and HLA-Cw*0303, respectively. We then identified a novel epitope FVYGGSKTSL presented by both HLA-Cw*0303 and Cw*0304, which are expressed by 40 % of Japanese. We made fluorescently labelled MHC-tetramers to detect EBNA1-specific CTLs by flow cytometry. The mixed lymphocyte-peptide culture method, which enabled us to estimate frequencies of EBNA1-specific CTLs, revealed that FVYGGSKTSL-specific CTL precursor frequencies in HLA-Cw*0303 or Cw*0304-positive donors were between 1 x 10^-5 and 1 x 10^-4 CD^8+ T cells. Moreover, both CTL clones inhibited growth of HLA-matched BEV-transformed B lymphocytes in vitro and B5 CTLs produced an IFN-y response to EBNA1-expressing gastric carcinoma cells in the context of HLA-Cw*0303.Accumulating current evidence indicates that CD^4+ T cells,as well as CTLs, are required for effective antitumor immunity. We investigated the ability of CD^4+. T cell clones induced with overlapping peptides covering EBNA1, and identified minimal epitopes and their restricted MEIC class II molecules. Of these, one clone recognized a novel epitope being presented by DRB1*0401, 0403, and 0406.

EB病毒（EBV）编码的核抗原I（EBNA 1）是针对EBV相关恶性肿瘤的免疫治疗的有吸引力的靶点，因为它在所有EBV阳性细胞中表达。尽管CD 48+细胞毒性T淋巴细胞（CTL）表位呈递在很大程度上被其独特结构域阻止，但使用mRNA转导的树突状细胞（DC）将提供引发EBNA 1特异性CTL的优点。在用EBNA 1转导的单核细胞衍生的DC刺激后，我们成功地从健康供体分离出两个EBNA 1特异性OTL克隆B5和C6。这些油分别识别HLA-B*3501和HLA-Cw*0303背景下的肽。然后，我们鉴定了由HLA-Cw*0303和Cw*0304呈递的新表位FVYGGSKTSL，其由40%的日本人表达。我们制作了荧光标记的MHC四聚体，通过流式细胞术检测EBNA 1特异性CTL。混合淋巴细胞-肽培养法使我们能够估计EBNA 1特异性CTL的频率，结果显示，在HLA-Cw*0303或Cw*0304阳性供体中，FVYGGSKTSL特异性CTL前体的频率在1 × 10^-5到1 × 10^-4 CD^8+ T细胞之间。此外，两个CTL克隆在体外抑制HLA匹配的BEV转化的B淋巴细胞的生长，并且B5 CTL在HLA-Cw* 0303的背景下对表达EBNA 1的胃癌细胞产生IFN-γ应答。我们研究了CD^4+的能力。用覆盖EBNA 1的重叠肽诱导T细胞克隆，并鉴定最小表位及其限制性MEIC II类分子。其中，一个克隆识别由DRB 1 *0401、0403和0406呈递的新表位。