Cyril Clavel
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Project: Macrophage polarization and expression of PADs, responsible for citrullination of autoantigen targeted by Rheumatoid Arthritis (RA)-specific anti-citrullinated protein autoantibodies (ACPA)
Autoantibodies to citrullinated proteins (ACPA) are specifically associated to RA and produced in inflamed RA synovial tissues (ST). In this ST, the peptidylarginine deiminases (PAD)2 and 4 are responsible for fibrin citrullination and therefore for genesis of the target-epitopes of the ACPA. PAD2 and 4 are expressed in the inflammatory infiltrate, essentially by CD68-positive mononuclear cells. Macrophages of the ST are therefore probably responsible for the synthesis and excretion of the enzymes in the synovial interstitium then of citrullination of the fibrin deposits.
By stimulating macrophages from healthy donors and from RA patients with ACPA-containing immune complexes (ACPA IC), we established their inflammatory potential. Furthermore, we set up a strategy allowing us to generate different human polarized macrophage subsets and demonstrated that the phenotype of macrophages greatly influences their cytokine responses to ACPA-IC.
Our project aim is now to evaluate the influence of such a polarization on the expression of the PAD2 and 4.
Autoantibodies to citrullinated proteins (ACPA) are specifically associated to RA and produced in inflamed RA synovial tissues (ST). In this ST, the peptidylarginine deiminases (PAD)2 and 4 are responsible for fibrin citrullination and therefore for genesis of the target-epitopes of the ACPA. PAD2 and 4 are expressed in the inflammatory infiltrate, essentially by CD68-positive mononuclear cells. Macrophages of the ST are therefore probably responsible for the synthesis and excretion of the enzymes in the synovial interstitium then of citrullination of the fibrin deposits.
By stimulating macrophages from healthy donors and from RA patients with ACPA-containing immune complexes (ACPA IC), we established their inflammatory potential. Furthermore, we set up a strategy allowing us to generate different human polarized macrophage subsets and demonstrated that the phenotype of macrophages greatly influences their cytokine responses to ACPA-IC.
Our project aim is now to evaluate the influence of such a polarization on the expression of the PAD2 and 4.
Animal and Cellular Models
Human macrophages differentiated or polarized in vitro from purified peripheral blood monocytes (from buffy coats )
Human macrophages differentiated or polarized in vitro from purified peripheral blood monocytes (from buffy coats )
Techniques and Methods:
CD14+-positive selection
Monocyte/macrophage culture without adherence
In vitro stimulation of macrophages by ACPA-immune complexes
CD14+-positive selection
Monocyte/macrophage culture without adherence
In vitro stimulation of macrophages by ACPA-immune complexes