Fig. 1: Development of CD153 vaccine.
a–c Seven-week-old C57BL/6J mice were immunized with the CD153-Alum vaccine (#a–e) or the KLH-Alum vaccine on days 0 and 14. a The titer against recombinant mouse CD153 is expressed as the OD at 450 nm at 28 days postimmunization. #a–e, n = 4, 6, 6, 3, 3, respectively; KLH, n = 3. b The antibody recognizes the rmCD153 protein by western blot analysis. As a negative control, 100 ng of rmOPN was applied to lane 1, while 30 and 100 ng of rmCD153 were applied to lanes 2 and 3, respectively. c The levels of IgG1 (Th2 response) and IgG2b, IgG2c, and IgG3 (Th1 response) in mice immunized with the CD153#D-Alum vaccine (n = 3) or the CD153#D-CpG vaccine (n = 6) on days 0, 14, and 28. Sera were collected on day 42. IgG1, p = 0.011; IgG2b, p = 0.0067; IgG2c, p = 0.0027; IgG3, p = 0.0029. Left; The titer against CD153-BSA is expressed as the half-maximal binding (OD 50%). Right; ratio of the titer against CD153-BSA-specific IgG2b:IgG1 (p = 0.022), IgG2c:IgG1 (p = 0.0091) and IgG3:IgG1 (p = 0.21). d The proportions of CD153+ senescent T cells in splenic tissues and VAT of male C57BL/6J mice at the ages of 20 and 3 months (normal diet). Senescent T cells were defined as PD-1+ CD153+ cells in CD4+, CD44hi, CD62Llo cells. e The proportion of senescent T cells induced by R848 administration in splenic tissues of male C57BL/6J mice (n = 6 in each group) at the ages of 18 weeks with or without the CD153-CpG vaccine or the KLH-CpG vaccine. PBS vs. R848, p = 0.0086; PBS vs. KLH-CpG, p = 0.0052; PBS vs. CD153-CpG, p = 0.98; R848 vs. KLH-CpG, p > 0.99; R848 vs. CD153-CpG, p = 0.020; KLH-CpG vs. CD153-CpG, p = 0.012. All the data are expressed as the mean ± SEM. Statistical evaluation was performed by one sided (a) and by two sided (c, e); analysis of variance (ANOVA) test (a); unpaired two-tailed t-test (c), natural logarithmic transformation; Tukey’s multiple comparison test e; *p < 0.05; **p < 0.01; ****p < 0.0001; ND, not detected.
