Mice were boosted day 0 and day 2 with i.p. of 3.54.0) with high frequency. The histological examination revealed severe infiltration of inflammatory cells in the spinal cord of MOG-immunized Wt mice while the resistance to EAE in GMF-KO mice was characterized by the absence of inflammatory cells. Administration of rGMF in Wt mice and GMF-KO mice resulted in a significant increase in infiltrating cells in the spinal cord following MOG-immunizations. We also evaluated cytokines and chemokines production as parameters of severity of inflammation in the spinal cord of Wt versus GMF-KO mice with and without GMF-reconstitution following MOG-immunizations. Cytokines (TNF-, IFN-, IL-1, IL-6) and chemokines (CCL2, CCL3, CXCL10, GM-CSF) production were significantly greater in Wt mice than in GMF-KO mice following MOG-immunization. Furthermore, the reconstitution experiment with rGMF showed that the administration of rGMF in both, Wt mice and GMF-KO mice produced significant increase in the GMF-mediated cytokine/chemokine production. Keywords:Glia maturation factor (GMF), Experimental autoimmune encephalomyelitis (EAE), Myelin oligodendrocyte glycoprotein 3555 (MOG 35-55), Cytokine/chemokine, Neuro inflammation == FX1 1. INTRODUCTION == Multiple sclerosis (MS) is a disabling, chronic relapsing inflammatory demyelinating disease of the central nervous system (CNS); and according to the National Multiple Sclerosis Society of USA, affecting 400,000 Americans and over two million individuals worldwide. Most people are diagnosed between the ages of 20 and 50, although individuals as young as 2 and as old as 75 have developed it. MS progresses in two phases; the earlier phase starts with an autoimmune inflammatory attack against myelin sheath components followed by a chronic phase of neuro-degeneration in which both the myelin sheath and the underlying axons are damaged (Steinman, 2001). The loss of axons and spinal cord atrophy result in paralysis in MS patients (Trapp et al., 1998). Since the pathogenesis of MS is not clear, no definitive treatment is yet available. Much of our current knowledge about contributing factors of MS is based on experimental autoimmune encephalomyelitis (EAE), an animal model with clinical and pathological similarities to MS (Martin et al., 1992,Steinman, 1996). Several theories for FX1 pathogenesis of MS implicate infiltrating T cells, pro-inflammatory cytokines, chemokines, antibody-mediated toxicity, activated macrophages, microglia and astrocytes (Cannella and Raine, 1995,Glabinski et al., 1999,Glabinski and Ransohoff, 1999b,a,Iglesias et al., 2001). Other inflammatory mediators implicated in EAE include highly reactive oxygen, nitrogen species, and nuclear transcription factor NF-kB (Smith et al., 1999,Baldwin, 2001). Despite significant advances, the mechanism by which autoimmune dysfunction results in tissue destruction in EAE remains unresolved. In the present study, we demonstrate for the first time that the administration of exogenous recombinant human GMF resulted in exacerbation of clinical symptoms of MOG-induced EAE in wild type mice. We also show, for the first time, that the delivery of exogenous recombinant human GMF could restored full-blown EAE in EAE-resistant GMF-deficient (GMF-KO) FX1 mice. We also show that the administration of exogenous recombinant human GMF enhanced proinflammatory cytokine/chemokine production in the CNS of MOG-immunized mice, suggesting that GMF accelerates progression of EAE by regulating GMF-mediated proinflammatory environment in the CNS of MOG-immunized mice. These observations extend the pathological role of GMF in the progression of EAE. == 2. EXPERIMENTAL PROCEDURES == == 2.1. Reagents == Myelin oligodendrocyte glycoprotein peptide 35-55 (MOG35-55), complete Freunds adjuvant and pertussis toxin were purchased from Sigma-Aldrich, St. Louis, MO. ELISA kits for mouse TNF- (Cat # KMC3011), IFN- (Cat # KMC4021), IL-1 (Cat # KMC0011), IL-6 (Cat # KMC0061), CCL2 (MCP-1, monocyte chemoattractant protein-1, Cat # KMC1011), CCL3 (MIP-1, macrophage inflammatory protein-1, Cat # KMC 2291), and GM-CSF (granulocyte-macrophage colony-stimulating factor, Cat # KMC2021) were obtained from FX1 BioSource International, CA. CXCL10 (IP-10, interferon gamma-induced protein FX1 10, Cat # ab100675) was from Abcam, Cambridge, MA. Recombinant human GMF (rGMF) was prepared essentially Rabbit Polyclonal to KAP1 as described earlier (Lim et al., 1989,Lim et al., 1990,Kaplan et al., 1991,Lim and Zaheer, 1991). == 2.2. GMF-deficient mice == GMF-deficient (GMF knockout) mice were originally produced by homologous recombination with over 80% of the amino acid sequence.