Effects of Ag85B stimulation and 4-1BB ligation on in vivo frequency of CD4+ and CD8+ T cells and 4-1BB expression of cells from NP-mice adoptively transferred into C57BL/6Ly5.1 recipient mice.

C57BL/6Ly5.1 mice adoptively transferred with 107 spleen cells from NP-mice (cells CD45.2+) were treated or not with antigen Ag85B and mAbs as described in Materials and Methods. Slpeen cells were harvested on post-infection day 10, stained with CD45.2-FITC, CD4-PE, CD8-PerCP, CD4PerCPCy5.5 and 4-1BB-PE mAbs and then analyzed by flow cytometry. The data, obtained by gating CD45.2+ cells on lymphocyte population, show the frequency of CD4+ and CD8+ T cells originating from NP-mice and the percentage of T cells expressing 4-1BB. The results are presented as mean ± SEM of 3 mice per group.#, § statistical significance for differences between groups determined by ANOVA and Bonferroni-type multiple t-test (# Ag85B/control IgG vs none; § Ag85B/control IgG vs Ag85B/anti-4-1BB mAb).