Characterisation of the immune response to type I collagen in scleroderma

Kenneth J Warrington, Usha Nair, Laura D. Carbone, Andrew H. Kang, Arnold E. Postlethwaite

Research output: Contribution to journalArticle

18 Citations (Scopus)

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. Tcell 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.

Original languageEnglish (US)
Article numberR136
JournalArthritis Research and Therapy
Volume8
Issue number4
DOIs
StatePublished - Jul 31 2006
Externally publishedYes

Fingerprint

Collagen Type I
T-Lymphocytes
Flow Cytometry
Cytokines
Interleukin-4
Interferons
Phenotype
Cell Line
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-10
Interleukin-2
Blood Cells
Rheumatoid Arthritis
Esters
Color
Tumor Necrosis Factor-alpha
Staining and Labeling
Antigens
Skin

ASJC Scopus subject areas

  • Rheumatology
  • Immunology
  • Immunology and Allergy

Cite this

Characterisation of the immune response to type I collagen in scleroderma. / Warrington, Kenneth J; Nair, Usha; Carbone, Laura D.; Kang, Andrew H.; Postlethwaite, Arnold E.

In: Arthritis Research and Therapy, Vol. 8, No. 4, R136, 31.07.2006.

Research output: Contribution to journalArticle

Warrington, Kenneth J ; Nair, Usha ; Carbone, Laura D. ; Kang, Andrew H. ; Postlethwaite, Arnold E. / Characterisation of the immune response to type I collagen in scleroderma. In: Arthritis Research and Therapy. 2006 ; Vol. 8, No. 4.
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