CTLA4-Ig interacts with cultured synovial macrophages from rheumatoid arthritis patients and downregulates cytokine production

Autor: Alberto Sulli, Stefano Soldano, Pierfranco Triolo, Paolo Clerico, Paola Montagna, Bruno Seriolo, Maurizio Cutolo, Lamberto Felli, Barbara Villaggio, Renata Brizzolara
Jazyk: angličtina
Předmět:
Male
Immunoconjugates
T-Lymphocytes
medicine.medical_treatment
Blotting
Western
Immunology
Antigen-presenting cells
Down-Regulation
Fluorescent Antibody Technique
Enzyme-Linked Immunosorbent Assay
chemical and pharmacologic phenomena
Proinflammatory cytokine
Abatacept
Arthritis
Rheumatoid
Rheumatology
Humans
Medicine
Cytotoxic T cell
Immunology and Allergy
Cells
Cultured
CD86
Reverse Transcriptase Polymerase Chain Reaction
business.industry
Cytotoxic T lymphocyte-associated antigen-4
Macrophages
Synovial Membrane
Interleukin
Macrophage Activation
Middle Aged
Immunohistochemistry
Molecular biology
Coculture Techniques
medicine.anatomical_structure
Cytokine
Antirheumatic Agents
CTLA-4
Disease-modifying antirheumatic drugs
Cytokines
Female
Tumor necrosis factor alpha
B7-2 Antigen
Synovial membrane
business
CD80
Research Article
Zdroj: Arthritis Research & Therapy
ISSN: 1478-6354
DOI: 10.1186/ar2865
Popis: Introduction Co-stimulatory signal B7(CD80/CD86):CD28 is needed in order to activate T cells in immune response. Cytotoxic T lymphocyte-associated antigen-4-immunoglobulin (CTLA4-Ig) binding to the B7 molecules on antigen-presenting cells downregulates this activation and represents a recent biological treatment in rheumatoid arthritis (RA). Objectives of the study were to investigate the presence of the B7.2 (CD86) molecule and its masking by CTLA4-Ig on cultures of both RA synovial macrophages (RA SM), and of macrophages differentiated from THP-1 cells (M). In addition, the anti-inflammatory effects of CTLA4-Ig on co-cultures of RA SM and M with activated T cells were tested. Methods All macrophages were co-cultured for 24 hours with activated T cells, without or with CTLA4-Ig (10, 100, 500 μg/ml for 1 hour, 3 hours and overnight, respectively). Immunofluorescence (IF) staining for B7.2, and an analysis of inflammatory cytokine expression (interleukin (IL) -6, tumor necrosis factor (TNF) α, IL-1β, transforming growth factor (TGF) β) by immunocytochemistry (ICC), western blot (WB) and reverse transcriptase-polymerase chain reaction (RT-PCR) were performed. Results Macrophages showed intense B7.2 expression. CTLA4-Ig/B7.2 masking was evident for all macrophages, even after only 1 hour of cell culture (range from 10 to 100 μg/ml). ICC of co-cultures showed a dose-dependent decrease in inflammatory cytokines (P < 0.001 for IL-6, TNFα, IL-1β and TGFβ). Data were confirmed by WB and RT-PCR analysis. Conclusions Optimal concentrations of CTLA4-Ig for the CTLA4-Ig/B7.2 masking on activated macrophages were identified and were found to induce significant downregulation in the cell production of IL-6, TNFα, IL1-β and TGFβ. In conclusion, macrophages would appear to be a sensitive target for CTLA4-Ig treatment in RA.
Databáze: OpenAIRE
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