Skip to main content

Advertisement

SpringerLink
Log in

Lactobacillus stress protein GroEL prevents colonic inflammation

  • Original Article—Alimentary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Feagins LA, Souza RF, Spechler SJ. Carcinogenesis in IBD: potential targets for the prevention of colorectal cancer. Nat Rev Gastroenterol Hepatol. 2009;6:297–305.

    Article  CAS  Google Scholar 

  2. Mowat AM, Agace WW. Regional specialization within the intestinal immune system. Nat Rev Immunol. 2014;14:667–85.

    Article  CAS  Google Scholar 

  3. Hooper LV, Macpherson AJ. Immune adaptations that maintain homeostasis with the intestinal microbiota. Nat Rev Immunol. 2010;10:159–69.

    Article  CAS  Google Scholar 

  4. Lozupone CA, Stombaugh JI, Gordon JI, et al. Diversity, stability and resilience of the human gut microbiota. Nature. 2012;489:220–30.

    Article  CAS  Google Scholar 

  5. Schwabe RF, Jobin C. The microbiome and cancer. Nat Rev Cancer. 2013;13:800–12.

    Article  CAS  Google Scholar 

  6. Kamada N, Seo SU, Chen GY, et al. Role of the gut microbiota in immunity and inflammatory disease. Nat Rev Immunol. 2013;13:321–35.

    Article  CAS  Google Scholar 

  7. Tuohy KM, Probert HM, Smejkal CW, et al. Using probiotics and prebiotics to improve gut health. Drug Discov Today. 2003;8:692–700.

    Article  Google Scholar 

  8. Ahl D, Liu H, Schreiber O, et al. Lactobacillus reuteri increases mucus thickness and ameliorates dextran sulphate sodium-induced colitis in mice. Acta Physiol (Oxf). 2016;217:300–10.

    Article  CAS  Google Scholar 

  9. Lamas B, Richard ML, Leducq V, et al. CARD9 impacts colitis by altering gut microbiota metabolism of tryptophan into aryl hydrocarbon receptor ligands. Nat Med. 2016;22:598–605.

    Article  CAS  Google Scholar 

  10. Oliva S, Di Nardo G, Ferrari F, et al. Randomised clinical trial: the effectiveness of Lactobacillus reuteri ATCC 55730 rectal enema in children with active distal ulcerative colitis. Aliment Pharmacol Ther. 2012;35:327–34.

    Article  CAS  Google Scholar 

  11. Isidro RA, Appleyard CB. Colonic macrophage polarization in homeostasis, inflammation, and cancer. Am J Physiol Gastrointest Liver Physiol. 2016;311:G59-73.

    Article  Google Scholar 

  12. Heinsbroek SE, Gordon S. The role of macrophages in inflammatory bowel diseases. Expert Rev Mol Med. 2009;11:e14.

    Article  Google Scholar 

  13. Rieu A, Aoudia N, Jego G, et al. The biofilm mode of life boosts the anti-inflammatory properties of Lactobacillus. Cell Microbiol. 2014;16:1836–53.

    Article  CAS  Google Scholar 

  14. Saibil H. Chaperone machines for protein folding, unfolding and disaggregation. Nat Rev Mol Cell Biol. 2013;14:630–42.

    Article  CAS  Google Scholar 

  15. Henderson B, Fares MA, Lund PA. Chaperonin 60: a paradoxical, evolutionarily conserved protein family with multiple moonlighting functions. Biol Rev Camb Philos Soc. 2013;88:955–87.

    Article  Google Scholar 

  16. Goloubinoff P, Gatenby AA, Lorimer GH. GroE heat-shock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli. Nature. 1989;337:44–7.

    Article  CAS  Google Scholar 

  17. Friedland JS, Shattock R, Remick DG, et al. Mycobacterial 65-kD heat shock protein induces release of proinflammatory cytokines from human monocytic cells. Clin Exp Immunol. 1993;91:58–62.

    Article  CAS  Google Scholar 

  18. Shin H, Jeon J, Lee JH, et al. Pseudomonas aeruginosa GroEL stimulates production of PTX3 by activating the NF-kappaB pathway and simultaneously downregulating microRNA-9. Infect Immun. 2017;85(3):e00935.

    Article  CAS  Google Scholar 

  19. Asea A. Chaperokine-induced signal transduction pathways. Exerc Immunol Rev. 2003;9:25–33.

    PubMed  PubMed Central  Google Scholar 

  20. Pais de Barros JP, Gautier T, Sali W, et al. Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay. J Lipid Res. 2015;56:1363–9.

    Article  CAS  Google Scholar 

  21. Menck K, Behme D, Pantke M, et al. Isolation of human monocytes by double gradient centrifugation and their differentiation to macrophages in teflon-coated cell culture bags. J Vis Exp. 2014;(19):e51554.

  22. Tsilingiri K, Barbosa T, Penna G, et al. Probiotic and postbiotic activity in health and disease: comparison on a novel polarised ex-vivo organ culture model. Gut. 2012;61:1007–15.

    Article  CAS  Google Scholar 

  23. Tsilingiri K, Sonzogni A, Caprioli F, et al. A novel method for the culture and polarized stimulation of human intestinal mucosa explants. J Vis Exp. 2013;(75):e4368.

  24. Gozzi GJ, Gonzalez D, Boudesco C, et al. Selecting the first chemical molecule inhibitor of HSP110 for colorectal cancer therapy. Cell Death Differ. 2020;27:117–29.

    Article  CAS  Google Scholar 

  25. Hayer-Hartl M, Bracher A, Hartl FU. The GroEL-GroES chaperonin machine: a nano-cage for protein folding. Trends Biochem Sci. 2016;41:62–76.

    Article  CAS  Google Scholar 

  26. Chassaing B, Aitken JD, Malleshappa M, et al. Dextran sulfate sodium (DSS)-induced colitis in mice. Curr Protoc Immunol. 2014;104:Unit 15 25.

  27. Rath E, Berger E, Messlik A, et al. Induction of dsRNA-activated protein kinase links mitochondrial unfolded protein response to the pathogenesis of intestinal inflammation. Gut. 2012;61:1269–78.

    Article  CAS  Google Scholar 

  28. Kim JJ, Shajib MS, Manocha MM, et al. Investigating intestinal inflammation in DSS-induced model of IBD. J Vis Exp. 2012;(60):3678.

  29. Siegemund S, Sauer K. Balancing pro- and anti-inflammatory TLR4 signaling. Nat Immunol. 2012;13:1031–3.

    Article  CAS  Google Scholar 

  30. Wirtz S, Neurath MF. Mouse models of inflammatory bowel disease. Adv Drug Deliv Rev. 2007;59:1073–83.

    Article  CAS  Google Scholar 

  31. Zeissig S, Rosati E, Dowds CM, et al. Vedolizumab is associated with changes in innate rather than adaptive immunity in patients with inflammatory bowel disease. Gut. 2018;68(1):25–39.

    Article  Google Scholar 

  32. Abreu MT. Toll-like receptor signalling in the intestinal epithelium: how bacterial recognition shapes intestinal function. Nat Rev Immunol. 2010;10:131–44.

    Article  CAS  Google Scholar 

  33. Gren ST, Grip O. Role of monocytes and intestinal macrophages in Crohn’s disease and ulcerative colitis. Inflamm Bowel Dis. 2016;22:1992–8.

    Article  Google Scholar 

  34. Arlet JB, Ribeil JA, Guillem F, et al. HSP70 sequestration by free alpha-globin promotes ineffective erythropoiesis in beta-thalassaemia. Nature. 2014;514:242–6.

    Article  CAS  Google Scholar 

  35. Seignez A, Joly AL, Chaumonnot K, et al. Serum Gp96 is a chaperone of complement-C3 during graft-versus-host disease. JCI Insight. 2017;2:e90531.

    Article  Google Scholar 

Download references

Acknowledgments

This work was funded by the “Ligue National contre le Cancer”, the “Fondation pour la Recherche Médicale” FDT20170436927 and by a French Government grant managed by the French National Research Agency under the program “Investissements d’Avenir” with reference ANR-11-LABX-0021 (LabEX LipSTIC), ANR-15-IDE and ANR-15-IDE-0003 (I-SITE-BFC). We thank the ”Conseil Regional de Bourgogne-Franche Comté” and the European Union program FEDER for their financial support, and "Cellimap Dijon" and the “Plateforme de cytométrie Dijon” for technical support.

Author information

Author notes

  1. Frédéric Lirussi and Carmen Garrido contributed to the co-direction of the work.

Authors and Affiliations

  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

    Alexandre M. M. Dias, Romain Douhard, François Hermetet, Mathilde Regimbeau, Tatiana E. Lopez, Daniel Gonzalez, Sophie Masson, Guillaume Marcion, Killian Chaumonnot, Burhan Uyanik, Sébastien Z. Causse, Tarik Hadi, Christelle Basset, Johanna Chluba, Jacques Grober, Pablo Ortega-Deballon, Oleg N. Demidov, Frédéric Lirussi & Carmen Garrido

  2. Faculty of Medicine and Pharmacy, Université de Bourgogne Franche-Comté, Dijon, France

    Alexandre M. M. Dias, Romain Douhard, François Hermetet, Mathilde Regimbeau, Tatiana E. Lopez, Daniel Gonzalez, Sophie Masson, Guillaume Marcion, Killian Chaumonnot, Burhan Uyanik, Sébastien Z. Causse, Tarik Hadi, Christelle Basset, Johanna Chluba, Jacques Grober, Fabrice Neiers, Pablo Ortega-Deballon, Oleg N. Demidov, Frédéric Lirussi & Carmen Garrido

  3. UMR PAM A 02.102, Dijon, France

    Aurélie Rieu & Jean Guzzo

  4. Centre des Sciences du Goût et de l’Alimentation, INRA, CNRS, Dijon, France

    Fabrice Neiers

  5. Institute of Cytology, RAS, St. Petersburg, Russia

    Oleg N. Demidov

  6. Centre Hospitalier Universitaire, Dijon, France

    Pablo Ortega-Deballon & Frédéric Lirussi

  7. Anticancer Centre Georges-François Leclerc (CGFL), Dijon, France

    Carmen Garrido

Authors
  1. Alexandre M. M. Dias
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Romain Douhard
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. François Hermetet
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mathilde Regimbeau
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tatiana E. Lopez
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Daniel Gonzalez
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sophie Masson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Guillaume Marcion
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Killian Chaumonnot
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Burhan Uyanik
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Sébastien Z. Causse
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Aurélie Rieu
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Tarik Hadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Christelle Basset
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Johanna Chluba
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Jacques Grober
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Jean Guzzo
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Fabrice Neiers
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Pablo Ortega-Deballon
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Oleg N. Demidov
    View author publications

    You can also search for this author in PubMed Google Scholar

  21. Frédéric Lirussi
    View author publications

    You can also search for this author in PubMed Google Scholar

  22. Carmen Garrido
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Carmen Garrido.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOC 4660 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Dias, A.M.M., Douhard, R., Hermetet, F. et al. Lactobacillus stress protein GroEL prevents colonic inflammation. J Gastroenterol 56, 442–455 (2021). https://doi.org/10.1007/s00535-021-01774-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00535-021-01774-3

Keywords

Search

Navigation