title: Milk fermentation products of L. helveticus R389 activate calcineurin as a signal to promote gut mucosal immunity.
abstract: BACKGROUND
  Fermented milks containing probiotic bacteria are a way of delivering bioactive constituents to targets in the gastrointestinal tract. We reported previously that the fermentation of milk at constant pH 6 by L. helveticus R389 increased its content of peptide fractions, and the oral administration of the non-bacterial fraction (FMSpH6) to mice increased total secretory IgA in the intestinal lumen and enhanced the number of IgA and various cytokines producing cells as well as the secretion of IL-6 by small intestine epithelial cells. We also demonstrated that this FMSpH6 was effective for the prevention of Salmonella typhimurium infection in mice. In this work, we studied in mice the impact of the oral administration of the supernatant of milk fermented by L. helveticus R389 on the gut physiology by measuring parameters such as calcium channels and E-cadherin expression, the activation of the biological signal calcineurin and mast and goblet cells, as a way to determine some mechanisms involved in the immunomodulating effects of the milk fermentation products, observed in previous studies. We analyzed the impact of the supernatant of milk fermented by L. helveticus R389 at pH6-controlled on the expression of calcineurin and on the reinforcement of the ephitelial barrier, measuring parameters such as calcium channels and E-cadherin expression and in the reinforcement of the non-specific immunity determining mast cells and goblet cells associated to the gut.
  
  
  RESULTS
  We observed an enhanced expression of TRPV6 channels in the duodenum, indicating an improved capacity for dietary Ca2+ uptake. We demonstrated an enhanced expression of calcineurin in the small intestine, able to upregulate immune parameters such as IL-2 and TNF production, with an increase in the number of these cytokines secreting cells. We determined an increase in the number of mucosal mast cells and goblet cells, which would mean an improved state of mucosal surveillance at sites of infection.
  
  
  CONCLUSIONS
  The oral administration of the supernatant of milk fermented by L. helveticus R389 enhanced the gut mucosal immunity by improving the mechanisms that reinforce the epithelial and non-specific barriers and the gut functioning at sites of infection, with an improvement in the expression of the enzyme calcineurin, an important signal in the network that activates the gut immune system. The results of this work contribute to revealing the mechanisms underlying the immunomodulation of the gut immune function by fermented milks with probiotic bacteria.
authors.affiliation: Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, Tucumán (4000), Argentina. gvinde@fiqus.unl.edu.ar
authors.forename: Gabriel
authors.initials: G
authors.lastname: Vinderola
authors.forename: Chantal
authors.initials: C
authors.lastname: Matar
authors.forename: Gabriela
authors.initials: G
authors.lastname: Perdigón
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D019703:Calcineurin / Q000378:metabolism*; D043302:Cultured Milk Products / Q000737:chemistry / Q000276:immunology* / Q000382:microbiology; D005285:Fermentation / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D020397:Goblet Cells / Q000378:metabolism; D018928:Immunity, Mucosal; D007376:Interleukin-2 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000276:immunology*; D052198:Lactobacillus helveticus / Q000502:physiology*; D008407:Mast Cells / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D019936:Probiotics; D050916:TRPV Cation Channels / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism