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. 2020 May 18;11:2482. doi: 10.1038/s41467-020-16347-w

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© The Author(s) 2020

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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Fig. 4 — a The proportions of senescent T cells induced by HFD loading in the VAT of mice immunized with the Alum-conjugated vaccine or the CpG-conjugated vaccine. Senescent T cells were defined as PD-1⁺ CD153⁺ cells in CD4⁺, CD44^hi, CD62L^lo cells. Alum-conjugated vaccine group, n = 3 in each group; CpG-conjugated vaccine group, n = 6 in each group; ND control group, n = 5; HFD control group, n = 6. b The CDC activity of the CD153 vaccine-induced antibody was assessed (n = 3 each sample) using 2 × 10⁴ LPS-stimulated RAW 264.7 cells. The vaccine-induced IgG antibodies used were purified from the sera of HFD-loaded mice at the age of 22–25 weeks immunized with the CD153-Alum vaccine, CD153-CpG vaccine, KLH-Alum vaccine or KLH-CpG vaccine. N, no stimulation with antibody; P, 100 µg/ml anti-mouse MHC class I antibody (H-2Kd/H-2Dd); closed right triangle, the concentrations of purified IgG antibodies; opened right triangle, the concentrations of anti-mouse CD153 antibody (10, 30, 100 µg/ml, respectively). c, d VAT were collected from the four groups of mice (ND control group, HFD control group, KLH-CpG vaccine group, and CD153-CpG vaccine group; n = 3 in each group) according to the time course of HFD loading and injection of vaccines. c Quantification of macrophages in the crown-like structures among the four groups. HFD vs. KLH-CpG, p = 0.11; HFD vs. CD153-CpG, p = 0.0041; KLH-CpG vs. CD153-CpG, p = 0.15. 40–50 fields per sample, F4/80 (brown); scale bars: 100 µm. d Quantification of CD153-positive cells in the crown-like structures among the four groups. HFD vs. KLH-CpG, p = 0.024; HFD vs. CD153-CpG, p = 0.0015; KLH-CpG vs. CD153-CpG, p = 0.20. 40–50 fields per sample, CD153 (brown); scale bars: 100 µm. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by two-sided; unpaired two-tailed t-test, KLH-Alum vs. CD153-Alum (a); Bonferroni correction, ND vs. HFD (a), HFD vs. KLH-CpG vs. CD153-CpG (a, c, d); *p < 0.05; **p < 0.01; n.s. not significant.