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Engineered and wild-type L. lactis promote anti-inflammatory cytokine signalling in inflammatory bowel disease patient’s mucosa

  • Saša Simčič
  • Aleš Berlec
  • Sanja StopinšekEmail author
  • Borut Štrukelj
  • Rok Orel
Short Communication

Abstract

Dysbiosis of intestinal microbiota and aberrant inflammatory responses in gastrointestinal mucosa plays important roles in the development of inflammatory bowel disease (IBD). The purpose of this study was to demonstrate the probiotic activity of Lactococcus lactis and the ability of TNF-α-binding by recombinant L. lactis bearing TNF-α-binding affibodies. Various concentrations of recombinant L. lactis were exposed to TNF-α and its binding measured by ELISA. Mucosal biopsies of patients with active IBD were incubated with various L. lactis strains or E. coli DH5α strain and concentrations of TNF-α, IL-23, and IL-10 in the supernatants determined by ELISA. Recombinant L. lactis, at 1 × 109 and 1 × 108 CFU/mL, bound 22.6% and 18.4%, respectively of TNF-α (p < 0.05). When IBD-mucosa was incubated with any L. lactis strain at 1 × 109 CFU/mL, levels of TNF-α and IL-23 were significantly decreased and that of IL-10 increased relative to that for the sterile culture. Opposite trends were observed with E. coli cultures. Recombinant L. lactis at 1 × 108 CFU/mL bound as much as 62.8% (p = 0.026) of TNF-α in IBD-mucosa supernatants compared with the control strain. L. lactis strains are reported, for the first time, to induce an ex vivo anti-inflammatory cytokine profile in IBD inflamed mucosa. L. lactis could therefore constitute a promising alternative approach for treating IBD.

Keywords

Cytokines Inflammatory bowel disease Lactococcus lactis Tumour necrosis factor-α 

Notes

Acknowledgements

We are grateful to Prof. Roger Pain for critical reading of the manuscript. This study was supported by a grant from the Slovenian Research Agency, Program Numbers P3-0083-0381 and P4-0127.

Compliance with ethical standards

Conflict of interest

Aleš Berlec and Borut Štrukelj are co-inventors of patent US 8754198 B2 (Modified food grade microorganism for treatment of inflammatory bowel disease), issued by United States Patent and Trademark Office. For the remaining authors, no conflicts of interest are declared.

Ethical approval

The National Medical Ethics Committee of the Republic of Slovenia approved the study protocol (Number 107/06/12). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. Askling J, Fahrbach K, Nordstrom B, Ross S, Schmid CH, Symmons D (2011) Cancer risk with tumor necrosis factor alpha (TNF) inhibitors: meta-analysis of randomized controlled trials of adalimumab, etanercept, and infliximab using patient level data. Pharmacoepidemiol Drug Saf 20:119–130.  https://doi.org/10.1002/pds.2046 CrossRefPubMedGoogle Scholar
  2. Baumgart DC, Sandborn WJ (2007) Inflammatory bowel disease: clinical aspects and established and evolving therapies. Lancet 369:1641–1657.  https://doi.org/10.1016/s0140-6736(07)60751-x CrossRefPubMedGoogle Scholar
  3. Baumgart DC, Sandborn WJ (2012) Crohn’s disease. Lancet 380:1590–1605.  https://doi.org/10.1016/s0140-6736(12)60026-9 CrossRefPubMedGoogle Scholar
  4. Braat H et al (2006) A phase I trial with transgenic bacteria expressing interleukin-10 in Crohn’s disease. Clin Gastroenterol Hepatol 4:754–759.  https://doi.org/10.1016/j.cgh.2006.03.028 CrossRefPubMedGoogle Scholar
  5. Cho JH (2008) The genetics and immunopathogenesis of inflammatory bowel disease. Nat Rev Immunol 8:458–466.  https://doi.org/10.1038/nri2340 CrossRefPubMedGoogle Scholar
  6. D’Ambrosio A et al (2016) Lamina propria CD4+ LAP+ regulatory T cells are increased in active ulcerative colitis but show increased IL-17 expression and reduced suppressor activity. J Crohns Colitis 10:346–353.  https://doi.org/10.1093/ecco-jcc/jjv216 CrossRefPubMedGoogle Scholar
  7. Danese S, Fiocchi C (2011) Ulcerative colitis. N Engl J Med 365:1713–1725.  https://doi.org/10.1056/NEJMra1102942 CrossRefPubMedGoogle Scholar
  8. Dianda L, Hanby AM, Wright NA, Sebesteny A, Hayday AC, Owen MJ (1997) T cell receptor-alpha beta-deficient mice fail to develop colitis in the absence of a microbial environment. Am J Pathol 150:91–97PubMedPubMedCentralGoogle Scholar
  9. Dignass A et al (2012) Second European evidence-based consensus on the diagnosis and management of ulcerative colitis part 1: definitions and diagnosis. J Crohns Colitis 6:965–990.  https://doi.org/10.1016/j.crohns.2012.09.003 CrossRefPubMedGoogle Scholar
  10. Ford AC, Peyrin-Biroulet L (2013) Opportunistic infections with anti-tumor necrosis factor-alpha therapy in inflammatory bowel disease: meta-analysis of randomized controlled trials. Am J Gastroenterol 108:1268–1276.  https://doi.org/10.1038/ajg.2013.138 CrossRefPubMedGoogle Scholar
  11. Garces S, Demengeot J, Benito-Garcia E (2013) The immunogenicity of anti-TNF therapy in immune-mediated inflammatory diseases: a systematic review of the literature with a meta-analysis. Ann Rheum Dis 72:1947–1955.  https://doi.org/10.1136/annrheumdis-2012-202220 CrossRefPubMedGoogle Scholar
  12. Gardlik R, Palffy R, Celec P (2012) Recombinant probiotic therapy in experimental colitis in mice. Folia Biol 58:238–245Google Scholar
  13. Jonsson A, Wallberg H, Herne N, Stahl S, Frejd FY (2009) Generation of tumour-necrosis-factor-alpha-specific affibody molecules capable of blocking receptor binding in vitro. Biotechnol Appl Biochem 54:93–103.  https://doi.org/10.1042/ba20090085 CrossRefPubMedGoogle Scholar
  14. Jostins L et al (2012) Host–microbe interactions have shaped the genetic architecture of inflammatory bowel disease. Nature 491:119–124.  https://doi.org/10.1038/nature11582 CrossRefPubMedPubMedCentralGoogle Scholar
  15. Khor B, Gardet A, Xavier RJ (2011) Genetics and pathogenesis of inflammatory bowel disease. Nature 474:307–317.  https://doi.org/10.1038/nature10209 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Kosler S, Strukelj B, Berlec A (2017) Lactic acid bacteria with concomitant IL-17, IL-23 and TNFalpha-binding ability for the treatment of inflammatory bowel disease. Curr Pharm Biotechnol 18:318–326.  https://doi.org/10.2174/1389201018666170210152218 CrossRefPubMedGoogle Scholar
  17. Kostic AD, Xavier RJ, Gevers D (2014) The microbiome in inflammatory bowel disease: current status and the future ahead. Gastroenterology 146:1489–1499.  https://doi.org/10.1053/j.gastro.2014.02.009 CrossRefPubMedPubMedCentralGoogle Scholar
  18. Kotlarz D et al (2012) Loss of interleukin-10 signaling and infantile inflammatory bowel disease: implications for diagnosis and therapy. Gastroenterology 143:347–355.  https://doi.org/10.1053/j.gastro.2012.04.045 CrossRefPubMedGoogle Scholar
  19. Lawson MM, Thomas AG, Akobeng AK (2006) Tumour necrosis factor alpha blocking agents for induction of remission in ulcerative colitis. Cochrane Database Syst Rev.  https://doi.org/10.1002/14651858.CD005112.pub2 CrossRefPubMedGoogle Scholar
  20. Leon AJ et al (2009) High levels of proinflammatory cytokines, but not markers of tissue injury, in unaffected intestinal areas from patients with. IBD Mediators Inflamm 2009:580450.  https://doi.org/10.1155/2009/580450 CrossRefPubMedGoogle Scholar
  21. Levine A et al (2014) ESPGHAN revised porto criteria for the diagnosis of inflammatory bowel disease in children and adolescents. J Pediatr Gastroenterol Nutr 58:795–806.  https://doi.org/10.1097/mpg.0000000000000239 CrossRefPubMedGoogle Scholar
  22. Llopis M et al (2009) Lactobacillus casei downregulates commensals’ inflammatory signals in Crohn’s disease mucosa. Inflamm Bowel Dis 15:275–283.  https://doi.org/10.1002/ibd.20736 CrossRefPubMedGoogle Scholar
  23. Lv R, Qiao W, Wu Z, Wang Y, Dai S, Liu Q, Zheng X (2014) Tumor necrosis factor alpha blocking agents as treatment for ulcerative colitis intolerant or refractory to conventional medical therapy: a meta-analysis. PLoS ONE 9:e86692.  https://doi.org/10.1371/journal.pone.0086692 CrossRefPubMedPubMedCentralGoogle Scholar
  24. McLean MH, Neurath MF, Durum SK (2014) Targeting interleukins for the treatment of inflammatory bowel disease-what lies beyond anti-TNF therapy? Inflamm Bowel Dis 20:389–397.  https://doi.org/10.1097/01.mib.0000437616.37000.41 CrossRefPubMedGoogle Scholar
  25. Nanda KS, Cheifetz AS, Moss AC (2013) Impact of antibodies to infliximab on clinical outcomes and serum infliximab levels in patients with inflammatory bowel disease (IBD): a meta-analysis. Am J Gastroenterol 108:40–47.  https://doi.org/10.1038/ajg.2012.363 (quiz 48) CrossRefPubMedGoogle Scholar
  26. Neurath MF (2014) Cytokines in inflammatory bowel disease. Nat Rev Immunol 14:329–342.  https://doi.org/10.1038/nri3661 CrossRefPubMedGoogle Scholar
  27. Olsen I, Lundin KE, Sollid LM (2013) Increased frequency of intestinal CD4+ T cells reactive with mycobacteria in patients with Crohn’s disease. Scand J Gastroenterol 48:1278–1285.  https://doi.org/10.3109/00365521.2013.837952 CrossRefPubMedPubMedCentralGoogle Scholar
  28. Orel R (2014) Intestinal microbiota, probiotics and prebiotics. Institute for Probiotics and Functional Foods Itd., LjubljanaGoogle Scholar
  29. Orel R, Kamhi Trop T (2014) Intestinal microbiota, probiotics and prebiotics in inflammatory bowel disease. World J Gastroenterol 20:11505–11524.  https://doi.org/10.3748/wjg.v20.i33.11505 CrossRefPubMedPubMedCentralGoogle Scholar
  30. Peyrin-Biroulet L, Deltenre P, de Suray N, Branche J, Sandborn WJ, Colombel JF (2008) Efficacy and safety of tumor necrosis factor antagonists in Crohn’s disease: meta-analysis of placebo-controlled trials. Clin Gastroenterol Hepatol 6:644–653.  https://doi.org/10.1016/j.cgh.2008.03.014 CrossRefPubMedGoogle Scholar
  31. Ravnikar M, Strukelj B, Obermajer N, Lunder M, Berlec A (2010) Engineered lactic acid bacterium Lactococcus lactis capable of binding antibodies and tumor necrosis factor alpha. Appl Environ Microbiol 76:6928–6932.  https://doi.org/10.1128/aem.00190-10 CrossRefPubMedPubMedCentralGoogle Scholar
  32. Sandborn WJ et al (2012) One-year maintenance outcomes among patients with moderately-to-severely active ulcerative colitis who responded to induction therapy with adalimumab: subgroup analyses from ULTRA 2. Aliment Pharmacol Ther 37:204–213CrossRefGoogle Scholar
  33. Sellon RK et al (1998) Resident enteric bacteria are necessary for development of spontaneous colitis and immune system activation in interleukin-10-deficient mice. Infect Immun 66:5224–5231PubMedPubMedCentralGoogle Scholar
  34. Siegel CA, Marden SM, Persing SM, Larson RJ, Sands BE (2009) Risk of lymphoma associated with combination anti-tumor necrosis factor and immunomodulator therapy for the treatment of Crohn’s disease: a meta-analysis. Clin Gastroenterol Hepatol 7:874–881.  https://doi.org/10.1016/j.cgh.2009.01.004 CrossRefPubMedPubMedCentralGoogle Scholar
  35. Sokol H et al (2008) Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patients. Proc Natl Acad Sci USA 105:16731–16736.  https://doi.org/10.1073/pnas.0804812105 CrossRefPubMedGoogle Scholar
  36. Song YN, Zheng P, Xiao JH, Lu ZJ (2014) Efficacy and safety of adalimumab for the Crohn’s disease: a systematic review and meta-analysis of published randomized placebo-controlled trials. Eur J Clin Pharmacol 70:907–914.  https://doi.org/10.1007/s00228-014-1702-1 CrossRefPubMedGoogle Scholar
  37. Stidham RW et al (2014a) Systematic review with network meta-analysis: the efficacy of anti-TNF agents for the treatment of Crohn’s disease. Aliment Pharmacol Ther 39:1349–1362.  https://doi.org/10.1111/apt.12749 CrossRefPubMedGoogle Scholar
  38. Stidham RW et al (2014b) Systematic review with network meta-analysis: the efficacy of anti-tumour necrosis factor-alpha agents for the treatment of ulcerative colitis. Aliment Pharmacol Ther 39:660–671.  https://doi.org/10.1111/apt.12644 CrossRefPubMedGoogle Scholar
  39. Taurog JD et al (1994) The germfree state prevents development of gut and joint inflammatory disease in HLA-B27 transgenic rats. J Exp Med 180:2359–2364CrossRefGoogle Scholar
  40. Turner D et al (2012) Management of pediatric ulcerative colitis: joint ECCO and ESPGHAN evidence-based consensus guidelines. J Pediatr Gastroenterol Nutr 55:340–361.  https://doi.org/10.1097/MPG.0b013e3182662233 CrossRefPubMedGoogle Scholar
  41. Verdier J, Begue B, Cerf-Bensussan N, Ruemmele FM (2012) Compartmentalized expression of Th1 and Th17 cytokines in pediatric inflammatory bowel diseases. Inflamm Bowel Dis 18:1260–1266.  https://doi.org/10.1002/ibd.21905 CrossRefPubMedGoogle Scholar
  42. Vitale S et al (2017) Cytokine production profile in intestinal mucosa of paediatric inflammatory bowel disease. PLoS ONE 12:e0182313.  https://doi.org/10.1371/journal.pone.0182313 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Institute of Microbiology and ImmunologyUniversity of Ljubljana, Faculty of MedicineLjubljanaSlovenia
  2. 2.Department of BiotechnologyJožef Stefan InstituteLjubljanaSlovenia
  3. 3.Faculty of PharmacyUniversity of LjubljanaLjubljanaSlovenia
  4. 4.Department of Gastroenterology, Hepatology and NutritionUniversity Children’s HospitalLjubljanaSlovenia

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