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. 2019 Dec 24;9:19767. doi: 10.1038/s41598-019-56365-3

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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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The role of T_FH cells in antibody productionand pathogenesis of AIHA. (A,B) CXCR5⁺CD4⁺CD25⁻T_FH cells and CXCR5⁻CD4⁺CD25⁻Tcells (non-T_FH) sorted from the responder group in week12 were cultured with CD19⁺ B cells from similarly immunized mice along with anti-CD3 and anti-IgM for six days. B cells were intracellularly stained for IgG1 (A) and surface stained for GL7 (B). Plots were pre-gated on CD19⁺ B cells. Each plot represented a single well. Data were from four independent experiments and the mean ± SEM were shown. ***p < 0.001. (C) The sorted CXCR5⁺CD4⁺CD25⁻T_FH cells and CXCR5⁻CD4⁻CD25⁻T cells from the responder group were adoptively transferred into C57BL/6J mice one day before rat RBCs injection on a weekly basis. Level of IgG-specific autoantibodies on mouse RBCs was measured by flow cytometry and expressed as a percentage of background unstained cells on week 2, 5, 8, and 12. Data shown are of three independent experiments with five mice in each group.