Dataset Information

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 ?-chain.

ABSTRACT:

Introduction

The V?12-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) ?-chain and consequently displays an increased immunity to CII and increased susceptibility to CIA. However, while the transgenic V?12 chain recombines with endogenous ?-chains, the frequency and distribution of CII-specific T cells in the V?12-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 V?12-transgenic mouse in order to determine to what extent the transgenic expression of the CII-specific ?-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 V?12-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 V?12-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.

SUBMITTER: Merky P

PROVIDER: S-EPMC2945055 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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Publications

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.

Merky Patrick P Batsalova Tsvetelina T Bockermann Robert R Dzhambazov Balik B Sehnert Bettina B Burkhardt Harald H Bäcklund Johan J

Arthritis research & therapy 20100803 4

<h4>Introduction</h4>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 ...[more]

PMID: 20682070

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Project description:INTRODUCTION: Interferon alpha (IFN-?) has a complex role in autoimmunity, in that it may both enhance and prevent inflammation. We have previously shown that the presence of IFN-? at sensitization protects against subsequent antigen-triggered arthritis. To understand this tolerogenic mechanism, we performed a descriptive, hypothesis-generating study of cellular and humoral responses associated with IFN-?-mediated protection against arthritis. METHODS: Arthritis was evaluated at day 28 in mice given a subcutaneous injection of methylated bovine serum albumin (mBSA), together with Freund adjuvant and 0 to 5,000 U IFN-? at days 1 and 7, followed by intraarticular injection of mBSA alone at day 21. The effect of IFN-? on mBSA-specific IgG1, IgG2a, IgG2b, IgA, and IgE was evaluated by enzyme-linked immunosorbent assay (ELISA). Cytokines in circulation and in ex vivo cultures on mBSA restimulation was evaluated with ELISA and Luminex, and the identity of cytokine-producing cells by fluorescence-activated cell sorting (FACS) analysis. RESULTS: Administration of IFN-? protected mice from arthritis in a dose-dependent manner but had no effect on antigen-specific antibody levels. However, IFN-? did inhibit the initial increase of IL-6, IL-10, IL-12, and TNF, and the recall response induced by intraarticular mBSA challenge of IL-1?, IL-10, IL-12, TNF, IFN-?, and IL-17 in serum. IFN-? decreased both macrophage and CD4+ T cell-derived IFN-? production, whereas IL-17 was decreased only in CD4+ T cells. Ex vivo, in mBSA-restimulated spleen and lymph node cell cultures, the inhibitory effect of in vivo administration of IFN-? on proinflammatory cytokine production was clearly apparent, but had a time limit. An earlier macrophage-derived, and stronger activation of the antiinflammatory cytokine transforming growth factor beta (TGF-?) was observed in IFN-?-treated animals, combined with an increase in CD4+ T cells producing TGF-? when arthritis was triggered by mBSA (day 21). Presence of IFN-? at immunizations also prevented the reduction in TGF-? production, which was induced by the intraarticular mBSA injection triggering arthritis in control animals. CONCLUSIONS: Administration of IFN-? has a profound effect on the cellular response to mBSA plus adjuvant, but does not affect antigen-specific Ig production. By including IFN-? at immunizations, spleen and lymph node cells inhibit their repertoire of antigen-induced proinflammatory cytokines while enhancing antiinflammatory TGF-? production, first in macrophages, and later also in CD4+ T cells. On intraarticular antigen challenge, this antiinflammatory state is reenforced, manifested as inhibition of proinflammatory recall responses and preservation of TGF-? levels. This may explain why IFN-? protects against antigen-induced arthritis.