Induction of proliferation and pro-inflammatory cytokine production in rheumatoid arthritis peripheral blood mononuclear cells by a 65 KDa chondrocyte membrane-specific, constitutive target autoantigen (CH65)
| Autor: | Joachim R. Kalden, S Alsalameh, Rayya J. Alsalameh, Gerd R Burmester, Jürgen Mollenhauer, Rachael C. Casey |
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| Rok vydání: | 2013 |
| Předmět: |
Adult
Male 0301 basic medicine Time Factors medicine.medical_treatment Ficoll Autoantigens Peripheral blood mononuclear cell Chondrocyte Proinflammatory cytokine Arthritis Rheumatoid 03 medical and health sciences Chondrocytes Immune system Rheumatology medicine Animals Humans Cells Cultured Aged Cell Proliferation Immunity Cellular business.industry Interleukin Middle Aged Molecular biology 030104 developmental biology Cytokine medicine.anatomical_structure Immunology Leukocytes Mononuclear Cytokines Cattle Female Tumor necrosis factor alpha Inflammation Mediators business |
| Zdroj: | International Journal of Rheumatic Diseases. 20:1132-1141 |
| ISSN: | 1756-1841 |
| DOI: | 10.1111/1756-185x.12167 |
| Popis: | Objective To analyze proliferation and pro-inflammatory cytokine production of peripheral blood mononuclear cells (PBMC) from rheumatoid arthritis (RA) patients following stimulation with a purified chondrocyte membrane-associated autoantigen (CH65). Methods CH65 was highly purified from bovine chondrocyte membranes by solubilization and ion exchange chromatography. PBMC of RA patients (n = 37; 28 seropositive, nine seronegative) and non-arthritic donors (n = 20) were isolated by ficoll centrifugation and used in cell proliferation assays. The levels of interleukin (IL)-1, tumo necrosis factor (TNF) and IL-6 produced after stimulation with CH65 were determined by enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed using Mann–Whitney U-test and Spearman rank test and the software SPSS 13.0TM (SPSS Inc.; Chicago, IL, USA). Results Peripheral blood mononuclear cells exhibited a strong proliferative response to purified CH65 in approximately 50% of the RA patients (seropositive > seronegative), with a maximum reactivity at 0.15 or 0.30 μg/mL culture medium. In contrast, PBMC from normal donors did not show a proliferative response to CH65 at any dose. The proliferative response in RA patients peaked at days 7–9 and returned to control levels at day 13, indicating an antigen-driven process. CH65-stimulated RA PBMC produced moderate to high amounts of IL-1, TNF and IL-6. This was comparable to the response after exposure to isolated whole chondrocyte membranes or purified collagen type II. Conclusion These results demonstrate a significant cellular immune response to CH65 protein in RA patients. Given the high similarity between bovine and human CH65, the results suggest a pathogenetic involvement of this molecule as a cartilage-specific potential target autoantigen in RA. |
| Databáze: | OpenAIRE |
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