title: Probiotic metabolites from Bacillus coagulans GanedenBC30™ support maturation of antigen-presenting cells in vitro.
abstract: OBJECTIVE
  To study the effects of probiotic metabolites on maturation stage of antigen-presenting immune cells.
  
  
  METHODS
  Ganeden Bacillus coagulans 30 (GBC30) bacterial cultures in log phase were used to isolate the secreted metabolite (MET) fraction. A second fraction was made to generate a crude cell-wall-enriched fraction, by centrifugation and lysis, followed by washing. A preparation of MET was subjected to size exclusion centrifugation, generating three fractions: < 3 kDa, 3-30 kDa, and 30-200 kDa and activities were tested in comparison to crude MET and cell wall in primary cultures of human peripheral blood mononuclear cell (PBMC) as a source of antigen-presenting mononuclear phagocytes. The maturation status of mononuclear phagocytes was evaluated by staining with monoclonal antibodies towards CD14, CD16, CD80 and CD86 and analyzed by flow cytometry.
  
  
  RESULTS
  Treatment of PBMC with MET supported maturation of mononuclear phagocytes toward both macrophage and dendritic cell phenotypes. The biological activity unique to the metabolites included a reduction of CD14+ CD16+ pro-inflammatory cells, and this property was associated with the high molecular weight metabolite fraction. Changes were also seen for the dendritic cell maturation markers CD80 and CD86. On CD14(dim) cells, an increase in both CD80 and CD86 expression was seen, in contrast to a selective increase in CD86 expression on CD14(bright) cells. The co-expression of CD80 and CD86 indicates effective antigen presentation to T cells and support of T helper cell differentiation. The selective expression of CD86 in the absence of CD80 points to a role in generating T regulatory cells.
  
  
  CONCLUSIONS
  The data show that a primary mechanism of action of GBC30 metabolites involves support of more mature phenotypes of antigen-presenting cells, important for immunological decision-making.
authors.affiliation: NIS Labs, 1437 Esplanade, Klamath Falls, OR 97601, USA. kathy@nislabs.com
authors.forename: Kathleen F
authors.initials: KF
authors.lastname: Benson
authors.forename: Kimberlee A
authors.initials: KA
authors.lastname: Redman
authors.forename: Steve G
authors.initials: SG
authors.lastname: Carter
authors.forename: David
authors.initials: D
authors.lastname: Keller
authors.forename: Sean
authors.initials: S
authors.lastname: Farmer
authors.forename: John R
authors.initials: JR
authors.lastname: Endres
authors.forename: Gitte S
authors.initials: GS
authors.lastname: Jensen
mesh_terms: D000328:Adult; D000938:Antigen-Presenting Cells / Q000187:drug effects* / Q000502:physiology; D018122:B7-1 Antigen / Q000032:analysis; D051940:B7-2 Antigen / Q000032:analysis; D001407:Bacillus / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D058851:GPI-Linked Proteins / Q000032:analysis; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008875:Middle Aged; D010586:Phagocytes / Q000166:cytology / Q000187:drug effects; D019936:Probiotics / Q000378:metabolism / Q000494:pharmacology*; D017452:Receptors, IgG / Q000032:analysis
chemical_list: D018122:B7-1 Antigen; D051940:B7-2 Antigen; C497964:CD86 protein, human; C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; D018950:Lipopolysaccharide Receptors; D017452:Receptors, IgG