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. 2021 Nov 19;12:747780. doi: 10.3389/fimmu.2021.747780

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Copyright © 2021 Tousif, Wang, Jackson, Hough, Strenkowski, Athar, Thannickal, McCusker, Ponnazhagan and Deshane

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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L-Kyn induced Breg differentiation was TLR-4 dependent but TLR-2 independent, the differentiation of Breg precursor was TLR-2 and TLR-4 independent. CD19⁺ B cells were negatively selected from whole splenocytes of TLR-4^-/-, TLR-2^-/- and TLR-2,4^-/- mice were disaggregated and seeded with LPS (10μg/ml), and increasing concentrations of L-Kyn (50µM and 100μM) for 72 hrs. Bregs were identified as CD19⁺CD5⁺CD1d^hiIL-10⁺ by flow cytometry. (A, C, E) Upper panel- L-Kyn induced Breg differentiation was impaired in B cells of TLR-4^-/- and TLR-2, 4^-/- but induction in TLR-2^-/- following stimulation with LPS+L-Kyn. Data shown here are pooled from four independent experiments with 3 technical replicates in each condition. (B, D, F) Lower panel- L-Kyn promoted Breg precursors (CD19⁺CD1d^hiCD5⁺) in B cells of TLR-4^-/- TLR-2^-/- and TLR-2, 4^-/- following stimulation with LPS+L-Kyn. Data shown here are pooled from three independent experiments with 3 technical replicates in each condition *p < 0.05, **p < 0.001, ***p < 0.0001, ****p < 0.00001.