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Arthritis Research and Therapy

, 8:R136

First Online: 31 July 2006Received: 21 February 2006Revised: 24 July 2006Accepted: 31 July 2006

Abstract

This study was conducted to examine the frequency, phenotype, and functional profile of T lymphocytes that proliferate in response to type I collagen CI in patients with scleroderma SSc. Peripheral blood mononuclear cells PBMCs from SSc patients, healthy controls, and rheumatoid arthritis disease controls were labeled with carboxy-fluorescein diacetate, succinimidyl ester CFSE, cultured with or without antigen bovine CI for 14 days, and analysed by flow cytometry. Surface markers of proliferating cells were identified by multi-color flow cytometry. T-cell lines were derived after sorting for proliferating T cells CFSE. Cytokine expression in CI-responsive T cells was detected by intracellular staining-flow cytometry and by multiplex cytokine bead assay Bio-Plex. A T-cell proliferative response to CI was detected in 8 of 25 32% SSc patients, but was infrequent in healthy or disease controls 3.6%; p = 0.009. The proliferating T cells expressed a CD4, activated CD25, memory CD45RO phenotype. Proliferation to CI did not correlate with disease duration or extent of skin involvement. T-cell lines were generated using in vitro CI stimulation to study the functional profile of these cells. Following activation of CI-reactive T cells, we detected intracellular interferon IFN-γ but not interleukin IL-4 by flow cytometry. Supernatants from the T-cell lines generated in vitro contained IL-2, IFN-γ, GM-CSF granulocyte macrophage-colony-stimulating factor, and tumour necrosis factor-α, but little or no IL-4 and IL-10, suggesting that CI-responsive T cells express a predominantly Th1 cytokine pattern. In conclusion, circulating memory CD4 T cells that proliferate to CI are present in a subset of patients with SSc, but are infrequent in healthy or disease controls.

AbbreviationsACR= American College of Rheumatology

APC= antigen-presenting cell

BSA= bovine serum albumin

CFSE= carboxy-fluorescein diacetate succinimidyl ester

CI= type I collagen

IFN-γ= interferon-γ

IL= interleukin

PBMC= peripheral blood mononuclear cell

PBS= phosphate-buffered saline

PE= phycoerythrin

SSc= scleroderma

TNF-α= tumour necrosis factor-α.

Electronic supplementary materialThe online version of this article doi:10.1186-ar2025 contains supplementary material, which is available to authorized users.

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Autor: Kenneth J Warrington - Usha Nair - Laura D Carbone - Andrew H Kang - Arnold E Postlethwaite

Fuente: https://link.springer.com/







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