Frédéric Lirussi1,2,3, Carmen Garrido4,5,6, Alexandre M M Dias1,2, Romain Douhard1,2, François Hermetet1,2, Mathilde Regimbeau1,2, Tatiana E Lopez1,2, Daniel Gonzalez1,2, Sophie Masson1,2, Guillaume Marcion1,2, Killian Chaumonnot1,2, Burhan Uyanik1,2, Sébastien Z Causse1,2, Aurélie Rieu7, Tarik Hadi1,2, Christelle Basset1,2, Johanna Chluba1,2, Jacques Grober1,2, Jean Guzzo7, Fabrice Neiers2,8, Pablo Ortega-Deballon1,2,3, Oleg N Demidov1,2,9. 1. INSERM, UMR 1231, Laboratoire d'Excellence LipSTIC and « Equipe labellisée par la Ligue Nationale Contre Le Cancer », 7 boulevard Jeanne d'Arc, 21079, Dijon, France. 2. Faculty of Medicine and Pharmacy, Université de Bourgogne Franche-Comté, Dijon, France. 3. Centre Hospitalier Universitaire, Dijon, France. 4. INSERM, UMR 1231, Laboratoire d'Excellence LipSTIC and « Equipe labellisée par la Ligue Nationale Contre Le Cancer », 7 boulevard Jeanne d'Arc, 21079, Dijon, France. cgarrido@u-bourgogne.fr. 5. Faculty of Medicine and Pharmacy, Université de Bourgogne Franche-Comté, Dijon, France. cgarrido@u-bourgogne.fr. 6. Anticancer Centre Georges-François Leclerc (CGFL), Dijon, France. cgarrido@u-bourgogne.fr. 7. UMR PAM A 02.102, Dijon, France. 8. Centre des Sciences du Goût et de l'Alimentation, INRA, CNRS, Dijon, France. 9. Institute of Cytology, RAS, St. Petersburg, Russia.
Abstract
BACKGROUND: We previously showed that supernatants of Lactobacillus biofilms induced an anti-inflammatory response by affecting the secretion of macrophage-derived cytokines, which was abrogated upon immunodepletion of the stress protein GroEL. METHODS: We purified GroEL from L. reuteri and analysed its anti-inflammatory properties in vitro in human macrophages isolated from buffy coats, ex vivo in explants from human biopsies and in vivo in a mouse model of DSS induced intestinal inflammation. As a control, we used GroEL purified (LPS-free) from E. coli. RESULTS: We found that L. reuteri GroEL (but not E. coli GroEL) inhibited pro-inflammatory M1-like macrophages markers, and favored M2-like markers. Consequently, L. reuteri GroEL inhibited pro-inflammatory cytokines (TNFα, IL-1β, IFNγ) while favouring an anti-inflammatory secretome. In colon tissues from human biopsies, L. reuteri GroEL was also able to decrease markers of inflammation and apoptosis (caspase 3) induced by LPS. In mice, we found that rectal administration of L. reuteri GroEL (but not E. coli GroEL) inhibited all signs of haemorrhagic colitis induced by DSS including intestinal mucosa degradation, rectal bleeding and weight loss. It also decreased intestinal production of inflammatory cytokines (such as IFNγ) while increasing anti-inflammatory IL-10 and IL-13. These effects were suppressed when animals were immunodepleted in macrophages. From a mechanistic point of view, the effect of L. reuteri GroEL seemed to involve TLR4, since it was lost in TRL4-/- mice, and the activation of a non-canonical TLR4 pathway. CONCLUSIONS: L. reuteri GroEL, by affecting macrophage inflammatory features, deserves to be explored as an alternative to probiotics.
BACKGROUND: We previously showed that supernatants of Lactobacillus biofilms induced an anti-inflammatory response by affecting the secretion of macrophage-derived cytokines, which was abrogated upon immunodepletion of the stress protein GroEL. METHODS: We purified GroEL from L. reuteri and analysed its anti-inflammatory properties in vitro in human macrophages isolated from buffy coats, ex vivo in explants from human biopsies and in vivo in a mouse model of DSS induced intestinal inflammation. As a control, we used GroEL purified (LPS-free) from E. coli. RESULTS: We found that L. reuteriGroEL (but not E. coliGroEL) inhibited pro-inflammatory M1-like macrophages markers, and favored M2-like markers. Consequently, L. reuteriGroEL inhibited pro-inflammatory cytokines (TNFα, IL-1β, IFNγ) while favouring an anti-inflammatory secretome. In colon tissues from human biopsies, L. reuteriGroEL was also able to decrease markers of inflammation and apoptosis (caspase 3) induced by LPS. In mice, we found that rectal administration of L. reuteriGroEL (but not E. coliGroEL) inhibited all signs of haemorrhagic colitis induced by DSS including intestinal mucosa degradation, rectal bleeding and weight loss. It also decreased intestinal production of inflammatory cytokines (such as IFNγ) while increasing anti-inflammatory IL-10 and IL-13. These effects were suppressed when animals were immunodepleted in macrophages. From a mechanistic point of view, the effect of L. reuteriGroEL seemed to involve TLR4, since it was lost in TRL4-/- mice, and the activation of a non-canonical TLR4 pathway. CONCLUSIONS:L. reuteriGroEL, by affecting macrophage inflammatory features, deserves to be explored as an alternative to probiotics.