Engineered and wild-type L. lactis promote anti-inflammatory cytokine signalling in inflammatory bowel disease patient’s mucosa
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.
KeywordsCytokines Inflammatory bowel disease Lactococcus lactis Tumour necrosis factor-α
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.
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 was obtained from all individual participants included in the study.
- 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
- Gardlik R, Palffy R, Celec P (2012) Recombinant probiotic therapy in experimental colitis in mice. Folia Biol 58:238–245Google Scholar
- 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
- 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
- Orel R (2014) Intestinal microbiota, probiotics and prebiotics. Institute for Probiotics and Functional Foods Itd., LjubljanaGoogle Scholar
- 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
- 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