Faculty of Pharmacy, Bezmialem Vakif University, Istanbul, Turkey; c Department of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, Istanbul, Turkey; d Department of Analytical Chemistry, Faculty of Pharmacy, Bezmialem Vakif University, Istanbul, Turkey The neuroinflammatory process plays a crucial role in the initiation and progression of multiple sclerosis (MS), and involves the activation of immune cells. During the neuroinflammatory process, release of proinflammatory mediators such as cytokines initiates the cascades of events leading into development of MS. The water extract of Capparis ovata (MSCov) which has been shown to be used as an alternative and complementary medicine for the treatment of MS, was further fractionated and studied for additional anti-neuroinflammatory effects in SH-SY5Y cells. In the present study, the butanolic fraction of the MSCov extract was tested for its anti-inflammatory effects on selected proinflammatory and inflam- matory genes believed to be important in MS pathophysiology using SH-SY5Y cells. In these cells, levels of the tumor necrosis factor-α (TNFα), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB1), glial fibrillary acidic protein (GFAP), C-X-C motif chemokine 10 (CXCL10), and tyrosine-protein phosphatase non- receptor type 11 (PTPN11) were determined by quantitative reverse transcriptase-PCR assay (qRT-PCR). Cell viability was assessed using lactate dehydrogenase (LDH) activity in the media conditioned by the crystal violet cell staining. We have found out that the butanolic fraction of MSCov significantly and effectively inhibited the expres- sion of all of the genes given above in SH-SY5Y cells. Thus, phytochemicals present in the butanolic subextract of the MSCov extract could be beneficial in preventing/treating neurodegenerative diseases in which neuroinflammation is part of the pathophysiology. Studies are underway to identify the individual compound(s) in this subextract of the MSCov extract contributing to these effects. This work is supported by TUBITAK 112S187. doi:10.1016/j.jneuroim.2014.08.464 117 Enhanced levels of IL-27 and IL-27R in the central nervous system of multiple sclerosis patients Vincent Sénécal a , Diane Beauseigle a , Raphael Schneider a , Craig Moore b , Alexandre Prat a , Jia Newcombe c , Samuel Ludwin d , Jack P. Antel b , Nathalie Arbour a a Research Center-CHUM, Université de Montréal, Montréal, Canada; b Montreal Neurological Institute, McGill University, Montréal, Canada; c UCL Institute of Neurology, University College London, London, United Kingdom; d Department of Neuropathology, Queen's University, Kingston, Canada Multiple sclerosis (MS) is pathologically characterized by focal demyelination, neuronal damage, and astrocyte and microglia activation. Multiple lines of evidence suggest that astrocytes and microglia can not only enhance immune mediated damages but they can also dampen inflammation and favor protection and repair. The mechanisms whereby CNS cells locally modulate immune responses are not fully elucidated. We have identified interleukin-27 (IL-27) as a key factor in the CNS inflammation. IL-27 is composed of two distinct subunits: EBI3 and p28 whereas the signaling receptor for IL- 27 (IL-27R) consists of two chains: T-cell cytokine receptor (TCCR (also named WSX-1)) and gp130. Both pro- and anti-inflammatory properties have been attributed to IL-27. Moreover, this cytokine has been shown to inhibit the development of experimental autoimmune encephalomyelitis, a mouse model of MS. However, whether IL-27 modulates the CNS inflammation observed in MS patients is still unknown. Our goal is to characterize the role of IL-27 in the immunopathological process of MS. We assessed the expression of IL- 27 (EBI3 and p28) and IL-27R (gp130 and TCCR) in post-mortem brain tissue samples. We observed that IL-27 is up-regulated in MS brains compared to controls and that astrocytes (GFAP+) and microglia/ macrophages (Iba1+) are important sources of this cytokine. We demonstrated that pro-inflammatory cytokines IFNgamma, IL-1beta, and TNF, up-regulate IL-27 production by primary cultures of human astrocytes as assessed by qPCR and ELISA. In addition, we showed that primary cultures of human pro-inflammatory macrophages and microglia (M1 cells) produce high levels of IL-27, whereas anti- inflammatory (M2 cells) do not. Moreover, we detected that most CNS infiltrating CD8 T cells express the receptor for IL-27, supporting the notion that these infiltrating immune cells are susceptible to the local IL-27-mediated effects. We have previously shown that IL-27 promotes the activation of human CD8 T cells into Tc1 cells carrying enhanced effector functions (IFN-gamma, granzyme B), thus CNS sources of IL-27 could locally increase cytotoxic function of these infiltrating T cells. Overall, our data demonstrate that IL-27 and its receptor are elevated in the CNS of MS patients and that inflammatory mediators present in MS lesions contribute to up-regulate such expression. Our future work will determine whether CNS IL-27 expression is beneficial or detrimental in the context of MS. doi:10.1016/j.jneuroim.2014.08.465 451 Proinflammatory monocyte production and distribution are modulated by alpha7 and alpha9 nicotinic acetylcholine receptors Alain Simard a , Wei Jiang b , Barbara Morley c a Department of Chemistry and Biochemistry and Centre de Formation Médicale du Nouveau-Brunswick, Université de Moncton, Moncton, Canada; b Neurology Department, Tianjin Medical University General Hospital, Tianjin, China; c Center for Hereditary Communication Disorders, Boys Town National Research Hospital, Omaha, United States Many studies have shown that nicotinic acetylcholine receptors (nAChRs) can modulate CNS and peripheral inflammation via the cholinergic anti-inflammatory pathway. Although it is clear that the alpha7 nAChR subtype is a key player in this process, increasing evidence suggests that other nAChRs may also modulate inflamma- tion. Previous data suggested that nicotine inhibits the production of monocytes and induces a shift towards anti-inflammatory mono- cyte/macrophage phenotype. However, whether nicotine modulates the production and distribution dynamics of pro-inflammatory Ly6C hi /CCR2 hi cells has not been studied to date. The goal of the present study was to determine if nicotine modulates the numbers of Ly6C hi /CCR2 hi in the bone marrow, blood, spleen and CNS of mice following immune stimulation. To this end, we assessed the proportions of Ly6C hi /CCR2 hi cells in these immune compartments 24 h after lipopolysaccharide (LPS) injection or at various time points (3, 7 and 16 days post-immunization) during the disease course of experimental autoimmune encephalomyelitis (EAE). We first found that nicotine prevents the increase in Ly6C hi /CCR2 hi cell numbers in the blood of mice 24 h after LPS injection and prior to EAE onset. This effect was lost in alpha7 nAChR knock-out mice, confirming that this receptor subtype plays an important role in nicotine's anti-inflammatory effects. Interestingly however, we observed a significant nicotinic effect on Abstracts 173