Increased prevalence of Methanosphaera stadtmanae in inflammatory bowel diseases

Abstract

Background: The gut microbiota is associated with the modulation of mucosal immunity and the etiology of inflammatory bowel diseases (IBD). Previous studies focused on the impact of bacterial species on IBD but seldom suspected archaea, which can be a major constituent of intestinal microbiota, to be implicated in the diseases. Recent evidence supports that two main archaeal species found in the digestive system of humans, Methanobrevibacter smithii (MBS) and Methanosphaera stadtmanae (MSS) can have differential immunogenic properties in lungs of mice; with MSS but not MBS being a strong inducer of the inflammatory response. We thus aimed at documenting the immunogenic potential of MBS and MSS in humans and to explore their association with IBD.

Methods: To validate the immunogenicity of MBS and MSS in humans, peripheral blood mononuclear cells from healthy subjects were stimulated with these two microorganisms and the production of inflammatory cytokine TNF was measured by ELISA. To verify MBS and MSS prevalence in IBD, stool samples from 29 healthy control subjects and 29 patients suffering from IBD were collected for DNA extraction. Plasma was also collected from these subjects to measure antigen-specific IgGs by ELISA. Quantitative PCR was used for bacteria, methanogens, MBS and MSS quantification.

Results: Mononuclear cells stimulated with MSS produced higher concentrations of TNF (39.5 ng/ml) compared to MBS stimulation (9.1 ng/ml). Bacterial concentrations and frequency of MBS-containing stools were similar in both groups. However, the number of stool samples positive for the inflammatory archaea MSS was higher in patients than in controls (47% vs 20%). Importantly, only IBD patients developed a significant anti-MSS IgG response.

Conclusion: The prevalence of MSS is increased in IBD patients and is associated with an antigen-specific IgG response.

Figure 1. TNF concentrations in supernatants of non-stimulated and MBS- or MSS-stimulated PBMC from healthy human subjects.

TNF was quantified by direct ELISA (average ± SEM). Cells from eleven subjects per group were analyzed. p<0.001.

Figure 2. IgG response in plasma of controls subjects and IBD patients.

Titers of A) MBS- and B) MSS-specific IgGs. C) Quantification of total IgGs. Results are expressed as average ± SEM. Eighteen controls (9 positive for MBS and 4 positive for MSS in stool samples) and seventeen patients (6 positive for MBS and 9 positive for MSS in stool samples) were analyzed. p<0.05.