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. 2019 Aug 29;9:12534. doi: 10.1038/s41598-019-47609-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Interleukin 1 beta (IL-1β) and active caspase-1 responses from peripheral blood mononuclear cells (PBMCs) were measured after stimulation with Mycobacterium avium complex (MAC) bacilli (multiplicity of infection [MOI] 20 and 100) and lipopolysaccharide (LPS) (5 µg/ml) for 48 hours. (A) The levels of secreted IL-1β production from the PBMCs after the indicated stimulation were measured using enzyme-linked immunosorbent assay (ELISA). (B) Intracellular IL-1β in the assay was detected using flow cytometry. We discriminated the lymphocytes and monocytes by forward scatter and side scatter. The percentages of IL-1β in CD14 + cells (upper panel) and CD4+ cells (lower panel) were gating in monocyte and lymphocyte populations, respectively. (C) The levels of intracellular IL-1β expressed on CD14⁺ monocytes were pooled by dot plots. (D) The levels of active caspase-1 from PBMCs assayed after MAC stimulation were measured by ELISA. The data were compared using the Mann Whitney U test. The crossed lines are mean values and bars of standard errors. *0.01 ≤ p < 0.05. **0.001 ≤ p < 0.01. ns, not significant; HC, healthy controls; pt, patients.