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MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) AS A POTENTIAL THERAPY FOR COLCHICINE NON-RESPONDERS OF FAMILIAL MEDITERRANEAN FEVER (FMF) PATIENTS


Authors: Ferial M. Hudaib, Ahmad R. Ramadan, Yousef Al-Abed and Said M. Shawar
Abstract: FMF is an autosomal recessive disorder highly prevalent around the Mediterranean Basin. Colchicine, a prophylactic drug for FMF, is effective in preventing FMF fever attacks (FA) in the majority of patients while 20%-30% of FMF patients are partially or totally non-responders to colchicine. MIF is a potent pro-inflammatory cytokine that plays an essential role in many inflammatory and autoimmune disorders. Blocking MIF tautomerase active site with small pharmacophores has been successfully used to suppress many autoimmune and inflammatory disorders. The role of MIF in FMF pathogenesis and the effect of MIF inhibition have not been evaluated. In this study, we investigated the effect of MIF reversible inhibitor ISO-1 on the expression of IL-1ß, TNF-a, IL-10 and MIF from peripheral blood lymphocytes (PBL) of FMF patients during FA. The ethics committee of our institution approved this study. ELISA assays and real time PCR (RT-PCR) were used to determine protein and mRNA expression levels of IL-1ß, TNF-a and IL-10 in FMF PBLs during FA before and after MIF inhibition. In addition, mRNA of MIF was determined under the same conditions in these patients. Our results were compared to age and sex matched healthy controls. Using ELISA assays, PBLs taken from FMF patients during FA, secrete 4-6 folds higher of IL-1ß, TNF-a and IL-10 compared to those from healthy matched controls. We determined a non-toxic optimal dose of 100uM and incubation time of 24hr for ISO-1 inhibition. ISO-1 significantly inhibited (50-70%) the secretion of IL-1ß, TNF-a and IL-10 to the media of cultured PBLs during FA. Interestingly, this inhibition returned the levels of assayed cytokines to that during remission. The mRNA level of IL-1ß, TNF-a, IL-10 and MIF was 1.5-2.0 folds higher during FA in FMF patients compared to healthy individuals. In addition, ISO-1 inhibited mRNA of all four cytokines between 30- 50% as determined by RT-PCR, after ISO-1 inhibition. To our knowledge, this is the first report investigating the effect of MIF inhibition on the expression of TNF-a, IL-1ß, Il-10 and MIF from PBLs of FMF patients. Different inhibition percentage of mRNA and protein secretion is likely due to different regulatory mechanisms employed during transcription and translation. Cytokine storm is the cause of fever, nausea and fatigue during immune reaction, i.e. FMF FA. The balance between pro- and anti-inflammatory cytokines usually determines the severity, duration and extent of inflammation. Thus, IL-10 elevation during FA is to counter the actions of TNF-a and IL-1ß and to prevent host tissue damages. Our results indicate that MIF is a major regulator of TNF-a, IL-1ß and IL-10 at the protein secretion and transcriptional level. We predict that this regulation is mediated by transcription pathway and transcriptional factors such as NF-kB. Taken together, our data suggests the possible use of ISO-1 as a therapeutic agent in treating FMF patients especially during the fever attacks.
Published in: The HUGO J;5(1):222
URL: http://www.cags.org.ae/4th pahgc. pdf
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