title: The JNk/NFkappaB pathway is required to activate murine lymphocytes induced by a sulfated polysaccharide from Ecklonia cava.
abstract: BACKGROUND
  The proven immunomodulatory and immune system activating properties of Ecklonia cava (E. cava) have been attributed to its plentiful polysaccharide content. Therefore, we investigated whether the sulfated polysaccharide (SP) of E. cava specifically activates the protein kinases (MAPKs) and nuclear factor-kappaB (NFkappaB) to incite immune responses.
  
  
  METHODS
  To assess immune responsiveness, lymphocytes were isolated from spleens of ICR mice and cultured with SP and its inhibitors. Assays included 3H-thymidine incorporation, flow cytometry, real time polymerase chain reaction (rtPCR), enzyme linked immunosorbent assay (ELISA), intracellular cytokine assay. Western blot, and electrophoretic mobility shift assay (EMSA).
  
  
  RESULTS
  SP dose-dependently increased the proliferation of lymphocytes without cytotoxicity. In particular, SP markedly enhanced the proliferation and differentiation of CD3+ mature T cells and CD45R/B220+ pan B cells. Additionally, SP increased the expression and/or production of IL-2, IgG(1a), and IgG(2b) compared to that in untreated cells. The subsequent application of JNK (SP600125), NFkappaB (PDTC), and serine protease (TPCK) inhibitors significantly inhibited the proliferation and IL-2 production of SP-treated lymphocytes as well as the phosphorylation of JNK and IkappaB, the activation of nuclear NFkappaB p65, and binding of NFkappaB p65 DNA. Moreover, co-application of both JNK and NFkappaB inhibitors completely blocked the proliferation of lymphocytes even in the presence of SP.
  
  
  CONCLUSIONS
  These results suggest that SP induced T and B cell responses via both JNK and NFkappaB pathways.
  
  
  CONCLUSIONS
  The effect of SP on splenic lymphocyte activation was assayed here for the first time and indicated the underlying functional mechanism.
authors.affiliation: Laboratory of Comparative Animal Medicine, Division of Animal Life Science, Institute of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
authors.forename: Ginnae
authors.initials: G
authors.lastname: Ahn
authors.forename: So Jin
authors.initials: SJ
authors.lastname: Bing
authors.forename: Sung-Myung
authors.initials: SM
authors.lastname: Kang
authors.forename: Won-Woo
authors.initials: WW
authors.lastname: Lee
authors.forename: Seung-Hong
authors.initials: SH
authors.lastname: Lee
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Matsuda
authors.forename: Akane
authors.initials: A
authors.lastname: Tanaka
authors.forename: Ik-Hyun
authors.initials: IH
authors.lastname: Cho
authors.forename: You-Jin
authors.initials: YJ
authors.lastname: Jeon
authors.forename: Youngheun
authors.initials: Y
authors.lastname: Jee
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D045424:Complex Mixtures / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D007376:Interleukin-2 / Q000096:biosynthesis / Q000276:immunology; D007795:Laminaria / Q000737:chemistry*; D008213:Lymphocyte Activation / Q000187:drug effects; D048670:MAP Kinase Kinase 4 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008813:Mice, Inbred ICR; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D010766:Phosphorylation / Q000187:drug effects; D011134:Polysaccharides / Q000302:isolation & purification / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D013936:Thymidine / Q000378:metabolism
chemical_list: D045424:Complex Mixtures; D007074:Immunoglobulin G; D007376:Interleukin-2; D016328:NF-kappa B; D011134:Polysaccharides; D047428:Protein Kinase Inhibitors; D048670:MAP Kinase Kinase 4; D013936:Thymidine