Characterisation of the immune response to type I collagen in scleroderma.

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 (CFSElow). 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)-gamma but not interleukin (IL)-4 by flow cytometry. Supernatants from the T-cell lines generated in vitro contained IL-2, IFN-gamma, GM-CSF (granulocyte macrophage-colony-stimulating factor), and tumour necrosis factor-alpha, 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.

• Authors: Kenneth J Warrington, Usha Nair, Laura D Carbone, Andrew H Kang, Arnold E Postlethwaite
• Journal: Arthritis research & therapy (Arthritis Res Ther) Vol. 8 Issue 4 Pg. R136 ( 2006) ISSN: 1478-6362 [Electronic] England
• PMID: 16879746 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• 5-(6)-carboxyfluorescein diacetate succinimidyl ester
• Antibodies
• Collagen Type I
• Cytokines
• Fluoresceins
• Fluorescent Dyes
• Succinimides

Topics

• Aged
• Antibodies (blood)
• CD4-Positive T-Lymphocytes (immunology, metabolism, pathology)
• Cell Division
• Cell Line
• Collagen Type I (immunology)
• Cytokines (metabolism)
• Female
• Flow Cytometry
• Fluoresceins
• Fluorescent Dyes
• Humans
• Immunologic Memory
• Male
• Middle Aged
• Scleroderma, Systemic (blood, immunology)
• Succinimides
• Th1 Cells (metabolism)
• Th2 Cells (metabolism)