title: In vitro and in vivo immunostimulatory activity of an exopolysaccharide-enriched fraction from Bacillus subtilis.
abstract: OBJECTIVE
  The aim of this study was to investigate the immunostimulatory effects of an exopolysaccharide-enriched fraction obtained from Bacillus subtilis J92 (B-EPS).
  
  
  RESULTS
  To determine the immunostimulatory activities of B-EPS, we used IFN-γ-primed RAW 264.7 macrophages and CD3/CD28-stimulated splenocytes. Increases in the levels of NO and many cytokines, such as, TNF-α, IL-6, and IL-1β, were observed in IFN-γ-primed RAW 264.7 macrophages by Griess reaction and ELISAs respectively. Using Western blotting and qRT-PCR, we found that B-EPS increased the protein and mRNA expressions of iNOS and the mRNA expressions of TNF-α, IL-6, and IL-1β. A reporter gene assay and EMSA revealed that B-EPS up-regulated the transcriptional activity of NF-κB by increasing its DNA binding and nuclear translocation. Pretreatment with NF-κB inhibitors, that is, BAY11-7082 and PDTC, decreased NO production in IFN-γ-primed RAW 264.7 macrophages by B-EPS. Furthermore, B-EPS increased the proliferation of and cytokine (IL-2 and IFN-γ) production by CD3/CD28-stimulated splenocytes. In a cyclophosphamide-induced immunosuppressed mouse model, B-EPS (5, 15 or 45 mg kg(-1) , p.o.) restored thymus and spleen indices. B-EPS also inhibited cyclophosphamide-induced reductions in neutrophil and lymphocyte numbers.
  
  
  CONCLUSIONS
  B-EPS improves immune function by regulating immunological parameters in IFN-γ-primed macrophages, CD3/CD28-stimulated splenocytes, and in cyclophosphamide-induced immunosuppressed mice.
  
  
  CONCLUSIONS
  This study suggests that the exopolysaccharides secreted by B. subtilis J92 could be used as immune stimulants.
authors.affiliation: Department of Pharmaceutical Biochemistry, College of Pharmacy, Kyung Hee University, Seoul, Republic of Korea; Department of Life and Nanopharmaceutical Science, Kyung Hee University, Seoul, Republic of Korea.
authors.forename: J-Y
authors.initials: JY
authors.lastname: Jung
authors.forename: J-S
authors.initials: JS
authors.lastname: Shin
authors.forename: Y K
authors.initials: YK
authors.lastname: Rhee
authors.forename: C-W
authors.initials: CW
authors.lastname: Cho
authors.forename: M-K
authors.initials: MK
authors.lastname: Lee
authors.forename: H-D
authors.initials: HD
authors.lastname: Hong
authors.forename: K-T
authors.initials: KT
authors.lastname: Lee
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D001412:Bacillus subtilis / Q000737:chemistry*; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D000276:Adjuvants, Immunologic; D016207:Cytokines; D015850:Interleukin-6; D016328:NF-kappa B; D011135:Polysaccharides, Bacterial; D014409:Tumor Necrosis Factor-alpha; C085953:exopolysaccharide, Bacillus; D052247:Nitric Oxide Synthase Type II
keywords: Bacillus subtilis; cyclophosphamide; exopolysaccharide; immunostimulatory activity; macrophage