Increased mucosal tumour necrosis factor alpha production in Crohn's disease can be downregulated ex vivo by probiotic bacteria.
OBJECTIVE
Tumour necrosis factor alpha (TNF-alpha) plays a key role in the pathogenesis of intestinal inflammation in Crohn's disease. The effect of bacteria on TNF-alpha release by intestinal mucosa was investigated.


METHODS
Ileal specimens were obtained at surgery from 10 patients with Crohn's disease (ileal stricture) and five disease controls undergoing right hemicolectomy (caecal cancer). Mucosal explants from each specimen were cultured for 24 hours with either non-pathogenic Escherichia coli, Lactobacillus casei DN-114001, L bulgaricus LB10, or L crispatus (each study contained blank wells with no bacteria). Tissue and bacterial viability was confirmed by lactate dehydrogenase (LDH) release and culture. Concentrations of TNF-alpha were measured in supernatants and the phenotype of the intestinal lymphocytes was analysed by flow cytometry.


RESULTS
Coculture of mucosa with bacteria did not modify LDH release. Release of TNF-alpha by inflamed Crohn's disease mucosa was significantly reduced by coculture with L casei or L bulgaricus; changes induced by L crispatus or E coli were not significant. The effect of L casei and L bulgaricus was not prevented by protease inhibitors. Coculture with L casei and L bulgaricus reduced the number of CD4 cells as well as TNF-alpha expression among intraepithelial lymphocytes from Crohn's disease mucosa. None of the bacteria induced changes in non-inflamed mucosa.


CONCLUSIONS
Probiotics interact with immunocompetent cells using the mucosal interface and modulate locally the production of proinflammatory cytokines.
Borruel
N
N
Digestive System Research Unit, Hospitals Vall d'Hebron, Autonomous University, Barcelona, Spain.
Carol
M
M
Casellas
F
F
Antolín
M
M
de Lara
F
F
Espín
E
E
Naval
J
J
Guarner
F
F
Malagelada
J R
JR
D000328:Adult; D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018920:Coculture Techniques; D003424:Crohn Disease / Q000178:diet therapy* / Q000276:immunology*; D046508:Culture Techniques; D004926:Escherichia coli; D005260:Female; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007778:Lactobacillus; D007780:Lacticaseibacillus casei; D018655:Lymphocyte Count; D008297:Male; D008875:Middle Aged; D019936:Probiotics / Q000008:administration & dosage*; D018709:Statistics, Nonparametric; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D014409:Tumor Necrosis Factor-alpha; D007770:L-Lactate Dehydrogenase