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. 2021 Feb 5;11:3222. doi: 10.1038/s41598-021-82930-w

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Radar plots showing the T_H1/T_H2/T_H17 like soluble cytokine profile. Cytokines were measured in supernatants of splenocyte cultures after 96 h of restimulation with Tc24-C4 protein (A–D) or T. cruzi Lysate (E–F) using a Luminex assay as described. Low dose vaccine + BZN (25 µg Tc24-C4/5 µg E6020 SE + 25 mg/kg BZN) significantly increased Tc24-C4 specific IL-17, IL-22, IL-23, IL-6, TNFα and IL-5 production when compared to infected untreated controls, and significantly increased IL-17 and TNFα when compared to low dose vaccine alone (A). In comparison, high dose vaccine + BZN induced significantly increased Tc24-C4 specific IL-23, IL-21, IL-6, IL-4, IL-5, IL-10 and TNFα when compared to infected untreated control mice, and decreased IL-17 and TNFα when compared to high dose vaccine alone (B) Similarly, low dose vaccine + BZN induced significantly increased Tc24-C4 specific IL-17, IL-22, IL-23, IL-6, TNFα and IL-5 production when compared to low dose BZN (C). In comparison , high dose vaccine + BZN induced significantly increased Tc24-C4 specific IL-23, IL-21, IL-6, IL-4, IL-5, IL-10 and TNFα when compared to low dose BZN (D). Low dose vaccine + BZN also induced significantly increased total parasite IL-21, IL-23, IL-10, IL-5, and IL-4 when compared to infected untreated controls, and increased IL-21, IL-4 and IL-5 compared to low dose vaccine alone (E). Low dose vaccine alone induced significantly increased IFNγ, IL-21, IL-23 and IL-10 compared to infected untreated controls (E). Similarly, high dose vaccine + BZN also induced significantly increased total parasite IFNγ, IL-10, IL-4, IL-21, Il-22, IL-23 and IL-17 compared to infected untreated controls, and increased IFNγ, IL-12, TNFα, IL-10, IL-4, IL-21, IL-23, and IL-17 when compared to high dose vaccine alone (F). High dose vaccine induced significantly increased IFNγ, TNFα and IL-22 compared to infected untreated mice. ^$Indicates a statistically significant difference (P value ≤ 0.05) when comparing low or high dose vaccine + BZN to infected untreated control mice (A,B,E,F), or when comparing low or high dose vaccine + BZN to low dose BZN alone (C and D). *Indicates a statistically significant difference (P value ≤ 0.05) when comparing low or high dose vaccine + BZN to vaccine alone (A and B), or when comparing low or high dose vaccine + BZN to low dose BZN alone (C and D). ^&Indicates a statistically significant difference (P value ≤ 0.05) when comparing low or high dose vaccine alone to infected untreated control mice (A,B,E,F), or when comparing low dose BZN to infected untreated controls (C and D).