在固定型 II 型胶原特异性转基因 T 细胞受体 β 链小鼠胶原诱导性关节炎中，促炎性抗原特异性 T 细胞的可视化和表型分析。

Visualization and phenotyping of proinflammatory antigen-specific T cells during collagen-induced arthritis in a mouse with a fixed collagen type II-specific transgenic T-cell receptor β-chain.

机构信息

Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.

出版信息

Arthritis Res Ther. 2010;12(4):R155. doi: 10.1186/ar3108. Epub 2010 Aug 3.

DOI:10.1186/ar3108

PMID:20682070

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2945055/

Abstract

INTRODUCTION

The Vβ12-transgenic mouse was previously generated to investigate the role of antigen-specific T cells in collagen-induced arthritis (CIA), an animal model for rheumatoid arthritis. This mouse expresses a transgenic collagen type II (CII)-specific T-cell receptor (TCR) β-chain and consequently displays an increased immunity to CII and increased susceptibility to CIA. However, while the transgenic Vβ12 chain recombines with endogenous α-chains, the frequency and distribution of CII-specific T cells in the Vβ12-transgenic mouse has not been determined. The aim of the present report was to establish a system enabling identification of CII-specific T cells in the Vβ12-transgenic mouse in order to determine to what extent the transgenic expression of the CII-specific β-chain would skew the response towards the immunodominant galactosylated T-cell epitope and to use this system to monitor these cells throughout development of CIA.

METHODS

We have generated and thoroughly characterized a clonotypic antibody, which recognizes a TCR specific for the galactosylated CII(260-270) peptide in the Vβ12-transgenic mouse. Hereby, CII-specific T cells could be quantified and followed throughout development of CIA, and their phenotype was determined by combinatorial analysis with the early activation marker CD154 (CD40L) and production of cytokines.

RESULTS

The Vβ12-transgenic mouse expresses several related but distinct T-cell clones specific for the galactosylated CII peptide. The clonotypic antibody could specifically recognize the majority (80%) of these. Clonotypic T cells occurred at low levels in the naïve mouse, but rapidly expanded to around 4% of the CD4+ T cells, whereupon the frequency declined with developing disease. Analysis of the cytokine profile revealed an early Th1-biased response in the draining lymph nodes that would shift to also include Th17 around the onset of arthritis. Data showed that Th1 and Th17 constitute a minority among the CII-specific population, however, indicating that additional subpopulations of antigen-specific T cells regulate the development of CIA.

CONCLUSIONS

The established system enables the detection and detailed phenotyping of T cells specific for the galactosylated CII peptide and constitutes a powerful tool for analysis of the importance of these cells and their effector functions throughout the different phases of arthritis.

摘要

简介

先前生成了 Vβ12 转基因小鼠，以研究抗原特异性 T 细胞在胶原诱导性关节炎（CIA）中的作用，CIA 是类风湿关节炎的动物模型。该小鼠表达一种转基因的 II 型胶原（CII）特异性 T 细胞受体（TCR）β链，因此对 CII 的免疫反应增强，对 CIA 的易感性增加。然而，尽管转基因 Vβ12 链与内源性α链重组，但 Vβ12 转基因小鼠中 CII 特异性 T 细胞的频率和分布尚未确定。本报告的目的是建立一种能够鉴定 Vβ12 转基因小鼠中 CII 特异性 T 细胞的系统，以确定转基因 CII 特异性β链的表达在何种程度上会使对免疫显性半乳糖化 T 细胞表位的反应产生偏差，并利用该系统在 CIA 发展过程中监测这些细胞。

方法

我们已经生成并彻底表征了一种克隆型抗体，该抗体识别 Vβ12 转基因小鼠中半乳糖化 CII（260-270）肽的 TCR。通过这种方式，可以定量和跟踪 CIA 发展过程中的 CII 特异性 T 细胞，并通过与早期激活标记物 CD154（CD40L）的组合分析以及细胞因子的产生来确定其表型。

结果

Vβ12 转基因小鼠表达几种相关但不同的 T 细胞克隆，这些克隆特异性针对半乳糖化的 CII 肽。克隆型抗体可以特异性识别其中的大多数（80%）。在未致敏的小鼠中，克隆型 T 细胞的水平较低，但迅速扩增到 CD4+T 细胞的 4%左右，此后随着疾病的发展，其频率下降。细胞因子谱分析显示，引流淋巴结中存在早期的 Th1 偏向性反应，在关节炎发作时会转向包括 Th17。数据表明，Th1 和 Th17 构成 CII 特异性群体中的少数，但表明调节 CIA 发展的抗原特异性 T 细胞的其他亚群。