Effect of colchicine and cytokines on MEFV expression and C5a inhibitor activity in human primary fibroblast cultures.
BACKGROUND
Familial Mediterranean fever is an autosomal recessive disease characterized by sporadic attacks of inflammation affecting the serosal spaces. The gene associated with FMF (MEFV), mainly expressed in neutrophils, was recently found to be expressed also in primary cultures of serosal origin (peritoneal and synovial fibroblasts). A C5a inhibitor, previously detected in normal serosal fluids, was recently identified in serosal cultures as well, and was found to be deficient in serosal fluids and cultures obtained from FMF patients.


OBJECTIVE
To investigate the effect of colchicine (the main therapeutic agent for FMF patients) and certain inflammatory cytokines (IL-1 beta, TNF-alpha, IFN-alpha, IFN-gamma) on MEFV expression and C5a inhibitor activity in neutrophils and primary peritoneal fibroblast cultures.


METHODS
Human primary peritoneal fibroblast cultures and neutrophils were studied for MEFV expression and C5a inhibitor activity, using reverse transcription-polymerase chain reaction and C5a-induced myeloperoxidase assay, respectively, in the presence and absence of colchicine and cytokines.


RESULTS
MEFV expression in neutrophils was high and could not be induced further. Its expression in the peritoneal fibroblasts was lower than in neutrophils and could be induced using colchicine and cytokines parallel with induction of C5a inhibitor activity. Semi-quantitative RT-PCR assays enabled estimation of MEFV induction by the cytokines at 10-100-fold and could not be further increased by concomitant addition of colchicine.


CONCLUSIONS
Serosal tissues, which are afflicted in FMF, express colchicine and cytokine-inducible MEFV and contain inducible C5a inhibitor activity. The relation between the ability of colchicine to induce MEFV and C5a inhibitor activity, and its efficacy in FMF treatment, require further investigation.
Abedat
Suzan
S
Hematology Unit, Hadassah Hospitals, Mount Scopus, Israel.
Urieli-Shoval
Simcha
S
Shapira
Eli
E
Calko
Sima
S
Ben-Chetrit
Eldad
E
Matzner
Yaacov
Y
D000970:Antineoplastic Agents / Q000494:pharmacology; D003078:Colchicine / Q000494:pharmacology*; D003169:Complement Inactivator Proteins / Q000187:drug effects* / Q000235:genetics*; D016207:Cytokines / Q000494:pharmacology*; D010505:Familial Mediterranean Fever / Q000235:genetics*; D005347:Fibroblasts / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects* / Q000235:genetics*; D006074:Gout Suppressants / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D016898:Interferon-alpha / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects*; D010537:Peritoneum / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000187:drug effects* / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000970:Antineoplastic Agents; D003169:Complement Inactivator Proteins; D016207:Cytokines; D006074:Gout Suppressants; D016898:Interferon-alpha; D007375:Interleukin-1; D014409:Tumor Necrosis Factor-alpha; C041833:complement C5a-inhibitors; D007371:Interferon-gamma; D012697:Serine Endopeptidases; D003078:Colchicine