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. 2017 Jul 19;7:5814. doi: 10.1038/s41598-017-06261-5

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© The Author(s) 2017

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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POL enhanced Tfh cell responses in immunized C57BL/6 mice. (A) 7 days after the 3^rd vaccination, the serum of immunized C57BL/6 mice were collected for HBV surface antibody level by using HBV surface antibody kit. (B) 7 days after the 3^rd vaccination, the splenocytes were stained with anti-mouse CD4-APC-Cy7/CXCR5-V450/PD-1-PE mAbs. The CD4⁺ cells were gated for Tfh cells analysis by flow cytometry. The CD4⁺CXCR5⁺ PD-1⁺ cells were counted relatively to total CD4⁺ cells. (C) The statistical results of Tfh cells were shown. (D) 7 days after the 3^rd vaccination, the splenocytes were stained with B220-PE-Cy5/CD95-APC/GL-7-FITC. The B220⁺ cells were gated for GC B cells analysis by flow cytometry. The B220⁺CD95⁺GL-7⁺ cells were counted relatively to total B220⁺ cells. (E) The statistical results of GC B cells were shown. Shown in each panel is 1 of at least 3 experiments with similar results. Bar, mean and SD from 3 independent experiments, each using at least three mice per group (n = 3); *p < 0.05; **p < 0.01.