Signal pathways in astrocytes activated by cross-talk between….pdfVIP

Kim et al. Journal of Neuroinflammation 2011, 8:25 JOURNAL OF /content/8/1/25 NEUROINFLAMMATION RESEARCH Open Access Signal pathways in astrocytes activated by cross-talk between of astrocytes and mast cells through CD40-CD40L * Dae Yong Kim, Gwan Ui Hong, Jai Youl Ro Abstract Background: Astrocytes, which play an active role in chronic inflammatory diseases like multiple sclerosis, exist close to mast cells with which they share perivascular localization. We previously demonstrated the possibility that astrocytes and mast cells interact in vitro and in vivo. This study aimed to investigate the signaling pathways and the role for astrocytes in the interaction of astrocytes and mast cells. Methods: We co-cultured human U87 glioblastoma (U87) and human mast cell-1 (HMC-1) cell lines, and mouse cerebral cortices-derived astrocytes and mouse bone marrow-derived mast cells (BMMCs). Intracellular Ca2+ ([Ca2+] ) i was measured by confocal microscopy; CD40 siRNA by Silencer Express Kit; small GTPases by GTP-pull down assay; PKCs, MAPKs, CD40, CD40L, Jak1/2, STAT1, TNF receptor 1 (TNFR1) by Western blot; NF-B and AP-1 by EMSA; cytokines by RT-PCR. An experimental allergic encephalomyelitis (EAE) model was induced using myelin oligodendrocyte glycoprotein (MOG) peptide and pertussis toxin in mice. Co-localization of TNFR1 and astrocytes in EAE brain tissues was determined by immunohistochemistry. Results: Each astrocyte co-culture had increases in [Ca2+]i levels, r