Analysis of food-antigen recognition by immune system using soluble T-cell receptor

使用可溶性 T 细胞受体分析免疫系统对食物抗原的识别

基本信息

批准号：
12660109
负责人：
TOTSUKA Mamoru
金额：
$ 0.9万
依托单位：
THE UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660109/
关键词：
T cell receptor β-Lactoglobulin Ovalbumin Food allergen Baculovirus expression system MHC class II molecule Antigen presentation Flow cytometry 食品アレルゲン昆虫細胞分泌発現腸管免疫抗原認識

项目摘要

This study aimed to analyze where food antigens are subjected to antigen presentation and recognized by T cells after ingested, using a soluble type of T cell receptor (TCR). The final aim of this study was to elucidate the mechanisms of immune recognition of food antigens. First, using β-lactoglobulin (β-Lg) and ovalbumin, primary food allergens, as model antigens, we analyzed the alteration of T cell functions when TCR-derived signals are altered by varying structure and dose of antigens used to stimulate the T cells. A soluble type of T cell receptor (sTCR) derived from a murine CD4^+ T cell clone specific for β-Lg, G1.19, was prepared using a baculovirus expression system. The sequence recognized by a biotinylation enzyme (BirA) was added downstream of the constant region of sTCR β chain. We confirmed that purified sTCR were biotinylated by BirA in the presence of biotin, and that multimer sTCR were formed by adding streptavidin. G1.19 cells recognize a peptide corresponding to 119-133 region of β-Lg(p119-133)in complex with I-A^b molecule. Proliferative response of G1.19 cells to the antigen was inhibited by adding sTCR to the culture, indicating that sTCR competitively inhibited the binding of TCR on G1.19 cells to the ligand. When the cells were stained with sTCR and fluorescence labeled anti-TCR Cβ antibodies in the presence of anti-MHC class II molecule, specific signals were detected by flow cytometry, indicating sTCR can specifically bind to the complex of the antigenic peptide and a MHC class II molecule.

这项旨在分析食物抗原的研究受到细胞受体类型后的T细胞识别（TCR）。 LG）和椭圆形的原发性食物过敏原，作为模型抗原，我们分析了R源性信号的改变，它通过用于刺激T细胞的抗原变化的结构和剂量改变了。对于β-LG，通过添加链链球菌的链球菌β链形成了β.19ressy系统。 -lg（P119-133）与I-A^B分子中的复合物。通过流式细胞仪检测II分子FIC信号，表明STCR可以特别结合抗原肽和MHC II类分子。

项目成果

期刊论文数量（51）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

戸塚護: "抗原アナログによるアレルギー応答の制御"臨床免疫. 36・5. 771-776 (2001)

Mamoru Totsuka：“抗原类似物控制过敏反应”临床免疫学36・5（2001）。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

戸塚護: "アレルギーの発症メカニズムと予防の考え方"FOOD Style 21. 5・3. 43-46 (2001)

户冢守：“过敏发生的机制及预防的思路” FOOD Style 21. 5・3. (2001)

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

戸塚護(分担執筆): "食品と免疫・アレルギー(上野川修一編)"朝倉書店. (2003)

户冢守（合著者）：《食物与免疫/过敏（上野川秀一编辑）》朝仓书店（2003）。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Wataru Ise, Mamoru Totsuka, Yoshitaka Sogawa, Akio Ametani, Satoshi Hachimura, Takehito Sato, Yoshihiro Kumagai, Sonoko Habu, and Shuichi Kaminogawa: "Naive CD4^+ T cells exhibit distinct expression patterns of cytokines and cell-surface molecules on thei

Wataru Ise、Mamoru Totsuka、Yoshitaka Sogawa、Akio Ametani、Satoshi Hachimura、Takehito Sato、Yoshihiro Kumagai、Sonoko Habu 和 Shuichi Kaminokawa：“幼稚 CD4^ T 细胞在其上表现出独特的细胞因子和细胞表面分子表达模式。