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A8.21 Rapid downregulation of inflammatory cytokine production in cultured human macrophages after CTLA4-Ig, dexamethasone, and methotrexate combined treatments
  1. M Cutolo,
  2. P Montagna,
  3. S Soldano,
  4. A Sulli,
  5. S Paolino,
  6. C Pizzorni,
  7. B Seriolo,
  8. M Cimmino,
  9. R Brizzolara
  1. Research Laboratory and Academic Division of Clinical Rheumatology, DIMI, University of Genova, Genova, Italy

Abstract

Background and objectives In rheumatoid arthritis (RA) the combination of the fusion protein CTLA4-Ig (abatacept) and other drugs such as glucocorticoids (GC) or/and methotrexate (MTX) allows to obtain better clinical improvement compared to CTLA4-Ig monotherapy. Our recent data showed that CTLA4-Ig, by interacting with the CD86 molecule on RA synovial macrophages, may induce reverse signalling with anti-inflammatory effects, involving NFkB intracellular pathway [1–4].

The anti-inflammatory effect of DEX alone or combined with CTLA4-Ig or/and with CTLA4-Ig plus methotrexate (MTX) was evaluated at gene expression level in cultured human activated macrophages.

Materials and methods THP-1 cells, activated into macrophages (PMA 0.05 g/ml; 24 h), were cultured for 3 and 24 h with DEX (10−8 M) alone, DEX combined with CTLA4-Ig (500 g/ml) and DEX with CTLA4-Ig plus MTX (0.05 g/ml). Subsequently, qRT-PCR analysis for IL-1b, TNFa and IL-6 gene expression was performed. Cells untreated were used as controls.

Results After 3 h, qRT-PCR showed in macrophages treated with DEX alone or with DEX-CTLA4-Ig or DEX-CTLA4-Ig-MTX combined treatment, a significant reduction (p < 0.01) for the expression of all assayed cytokines, compared with controls. CTLA4-Ig alone after 3 hrs, also significant reduced IL-1b (p < 0.01), TNFa (p < 0.05) and IL-6 (p < 0.01) expression. After 24 h, DEX alone or DEX-CTLA4-Ig or DEX-CTLA4-Ig-MTX combined treatments still showed the most significant reduction (p < 0.01) only for IL-1b. After 24 h of DEX-CTLA4-Ig or DEX-CTLA4-Ig-MTX combined treatments TNFa and IL-6 gene expression were still decreased. On the contrary, TNFa and IL-6 gene expression after 24 h of DEX alone treatment resulted unchanged, compared to controls. CTLA4-Ig alone, after 24 h also induced a significant decrease in gene expression for TNFa (p < 0.05) and IL6 (ns), while IL1b expression was unchanged, compared to controls.

Conclusions DEX and DEX-CTLA4-Ig or DEX-CTLA4-Ig-MTX combined, induced a rapid anti-inflammatory effect on cultured human macrophages, by decreasing proinflammatory cytokine gene expression already after 3 h of treatment. These results seem well correlated with the clinical experience.

References

  1. Cutolo M, Soldano S, Montagna P, et al . Arthritis Res Ther 2009; 11 :176–85.

  2. Brizzolara R, Soldano S, Montagna P, et al . Reumatismo 2011; 63 :80–5.

  3. Brizzolara R, Montagna P, Soldano S, Cutolo M. J Rheumatol 2013; 40 (5):738–40.

  4. Cutolo M, Soldano S, Contini P, et al . Clin Exp Rheumatol 2013; 31 (6):943–6.

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