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. 2020 Oct 30;10:573813. doi: 10.3389/fcimb.2020.573813

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Copyright © 2020 Reis, Dupin, Costa, Toledo, de Oliveira, Popi, Torrecilhas and Xander

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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Expression of IL-6, IL-10, and TNF-α in B-1 cells after stimulation for 24 or 48 h with L. amazonensis promastigotes (left panel) or EVs (right panel). (A) and (B) stimulation with the parasites for 24 or 48 h, respectively; (C) 24 and (D) 48 h of stimulation with EVs. B-1 cells were enriched using negative selection with anti-CD23 microbeads and positive selection with anti-CD19 microbeads. Animals inoculated with PBS were used as control. The RNA was extracted, the cDNA obtained, and qPCR analyses were performed to verify the gene expression of IL-6, IL-10, and TNF-α cytokines. The bars indicate the average of triplicates, and the error bars the standard deviation. The graph is representative of three independent experiments. Test t-student, *P < 0.05; ***P < 0.001; ****P < 0.0001, comparing stimulated (with EVs or parasites) and unstimulated B-1 cells.