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. 2018 Nov 14;9:2656. doi: 10.3389/fimmu.2018.02656

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Copyright © 2018 Dumas, Corral, Colom, Levillain, Peixoto, Hudrisier, Poquet and Neyrolles.

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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MAIT cell accumulation in the lungs and IL-17A-production are decreased early after M. tuberculosis infection in microbiota-altered mice. (A) Gating strategy to analyze MAIT cells (MR1-5-OP-RU tetramer⁺TCRβ⁺) by flow cytometry. A MR1–Ac-6-FP tetramer⁺TCRβ⁺ staining was used as a control. (B,C) Total number (B) and percentage (C) of MAIT cells in the lungs of control (non-ABX) ABX, and FT mice 0 and 7 (B, left panel), 14 and 21 (B, right panel), and 7 (C) days p.i., Data from 2–6 independent experiments (n = 4–8 mice/group/experiment) were pooled and the graphs show median with interquartile range of the pooled data. Data were analyzed using the Kruskal-Wallis test. (D) Intracellular analysis of Ki67 expression in MAIT cells from non-ABX, ABX, and FT mice 7 days p.i., by flow cytometry. Data from 2 independent experiments (n = 4–8 mice/group/experiment) were pooled and the graphs show the median with interquartile range of the pooled data. Data were analyzed using the Kruskal-Wallis test. (E) The expression of CCR6 (left) in whole lung homogenates or in MAIT cells (right), and the production of CCL20 in lung supernatant (middle), were measured by RT-qPCR (left), flow cytometry (right) and ELISA (middle) in ABX and control (non-ABX) mice 7 days p.i., RT-qPCR data were analyzed as in Supplementary Figure 2. Data from 3 (left) or 4 (middle and right) independent experiments (n = 3–7 mice/group/experiment) were pooled and the graphs show mean ± SD of the pooled data. Data were analyzed using the Student's t-test. (F) Cytometry analysis of percentage of activated (MR1-5-OP-RU tetramer⁺TCRβ⁺CD69⁺) MAIT cells (left), and IL-17A-producing (MR1-5-OP-RU tetramer⁺TCRβ⁺IL-17A⁺) MAIT cells (right) in lungs of control (non-ABX), ABX and FT mice 7 days p.i., Data from 4 (left) or 1 representative of 3 (right) independent experiments (n = 5–8 mice/group/experiment) were pooled and the graphs show mean ± SD of the pooled data. (left) or median with interquartile range (right) of the pooled data. Data were analyzed using the unpaired Student's t-test (left) or the Kruskal-Wallis test (right). NS, not significant; *p < 0.05; **p < 0.01.