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. 2020 Apr 22;11:755. doi: 10.3389/fmicb.2020.00755

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Copyright © 2020 Krause, Schaepe, Schattenberg, Müller, Ackermann, Rolle-Kampczyk, Jehmlich, Pierzchalski, von Bergen and Herberth.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Stimulation of mucosal-associated T (MAIT) cells with individual community members of SIHUMIx. (A) We identified MAIT cells as alive CD3⁺, CD8⁺, CD161⁺, and TCRvα7.2⁺ from single cell lymphocytes. (B) We defined activated MAIT cells by the expression of CD69⁺/TNF⁺ and show exemplary dot plots of unstimulated MAIT cells and MAIT cells after stimulation with the individual SIHUMIx strains [25 bacteria per cell (BpC)]. Moreover, we mixed the individual SIHUMIx strains in equal cell number (SIHUMIx1:1 mix) for stimulation (25 BpC). (C) Bar plots quantify the percentage of activated CD69⁺/TNF⁺ MAIT cells after stimulation with the individual SIHUMIx strains compared to the unstimulated control. Bars represent mean ± sd, n = 6, ordinary one-way ANOVA, unpaired, Dunnett’s multiple comparison test. The riboflavin (D) and folate (E) concentrations were measured in the culture supernatant of SIHUMIx after cultivation in Brain-Heart Infusion (BHI) medium [ng/mL]. Bars represent mean ± sd, n = 3. Positive changes indicate vitamin secretion, negative values indicate vitamin uptake.