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. 2018 Oct 10;8:15102. doi: 10.1038/s41598-018-33253-w

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© The Author(s) 2018

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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Systemic immune responses in tumor-bearing and healthy mice. Levels of anti-BCG (a) and anti-M. brumae (b) IgG antibodies present in sera from BCG-treated, M. brumae-treated and non-treated tumor-bearing mice, as well as in healthy animals. The presence of BCG antibodies in M. brumae-treated mouse serum is represented by empty blue symbols (b) and the presence of M. brumae antibodies in BCG-treated mouse serum by empty red symbols (a). For a and b, solid lines represent the mean of the OD (405 nm) values (two technical replicates) of the six mice from each group. *p < 0.05 compared to No-bact. ^&p < 0.05 compared to H. Splenocyte proliferation after ex vivo BCG (c) and M. brumae (d) restimulation. Proliferation is expressed relative to non-restimulated splenocytes of the same animal (proliferation of restimulated splenocytes/proliferation of non-restimulated splenocytes) after BCG and M. brumae stimulation. Proliferation of splenocytes from M. brumae-treated animals restimulated with BCG antigens is represented by empty red symbols (c) and proliferation of splenocytes from BCG-treated animals restimulated with M. brumae antigens is represented by empty blue symbols (d). For c and d, solid lines represent the mean of relative proliferation (three technical replicates) of 3–6 different spleens from each animal group. *p < 0.05 compared to No-bact. ^&p < 0.05 compared to H. For a, b, c, and d, dots represent animals treated with emulsified preparations and triangles represent animals treated with non-emulsified preparations. The different groups of animals are indicated by different colors on the horizontal axis of the graph as follows: empty grey triangles for the healthy (H) mice, empty black symbols for tumor-bearing mice receiving no mycobacteria treatment (No-bact), blue symbols for BCG-treated mice, and red symbols for M. brumae-treated mice.