impact factor, citescore
logo
In press
In Press
Recent issue
Recent issue
Recent Supplement
Recent supplement
archives
Archives
articles free
Free to view
Search
Search
 

Full Papers

 

Effects of combined treatments with CTLA4-IG (abatacept), dexamethasone and methotrexate on cultured human macrophages

1, 2, 3, 4, 5, 6, 7, 8, 9, 10

 

  1. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy. mcutolo@unige.it
  2. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.
  3. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.
  4. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.
  5. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.
  6. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.
  7. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.
  8. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.
  9. Division of Clinical Immunology, Department of Internal Medicine, University of Genova, Italy.
  10. Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Italy.

CER8875
2016 Vol.34, N°3
PI 0500, PF 0506
Full Papers

Free to view
(click on article PDF icon to read the article)

PMID: 26940199 [PubMed]

Received: 19/08/2015
Accepted : 14/12/2015
In Press: 03/03/2016
Published: 30/05/2016

Abstract

OBJECTIVES:
To evaluate the anti-inflammatory effect of CTLA4-Ig (abatacept) and dexamethasone (DEX) monotreatment versus their combination and adding methotrexate (MTX) on cultured human macrophages.
METHODS:
THP-1 cells, activated into macrophages (PMA 0.05 μg/ml), were cultured for 3 and 24 hrs with CTLA4-Ig (500 μg/ml), DEX (10-8 M), MTX (0.05 μg/ml), and CTLA4-Ig combined with DEX or CTLA4-Ig combined with DEX plus MTX. CTLA4-Ig/CD86 interaction was evaluated by FACS analysis. Quantitative real time-PCR (qRT-PCR), immunocytochemistry (ICC) and immunoassay (ELISA) analysis for inflammatory cytokine (IL-1β, TNF-α, IL-6) expression were performed.
RESULTS:
FACS analysis showed in macrophages treated with CTLA4-Ig alone, CTLA4-Ig-DEX and CTLA4-Ig-DEX-MTX a CD86 decrease of almost 35%, versus untreated cells (CNT). After 3 hrs, macrophages treated with DEX alone or with CTLA4-Ig-DEX or CTLA4-Ig-DEX-MTX showed a significant reduction (p<0.05) for all cytokines gene expression, that was still significant for IL-1β after 24 hrs (p<0.05). After 3 hrs, CTLA4-Ig alone significantly (p<0.05) reduced all cytokine genes; however, after 24 hrs still evident only for TNF-α (p<0.05). After 24 hrs CTLA4-Ig-DEX induced a significant decrease of gene expression (p<0.05) for TNF-α and IL-6, whereas CTLA4-Ig-DEX-MTX induced a decrease (p<0.05) limited to IL-6, versus CNT. Finally, ICC showed, after 24 hrs of CTLA4-Ig-DEX or CTLA4-Ig-DEX-MTX treatment a reduction (p<0.05) of IL-1β and IL-6 expression, versus CNT; DEX alone reduced only IL-1β (p<0.05). ELISA analysis confirmed these results.
CONCLUSIONS:
CTLA4-Ig-DEX and CTLA4-Ig-DEX-MTX combined treatments, decreased at any level the inflammatory cytokine expression more efficiently then monotreatments on activated cultured human macrophages.

Rheumatology Article