Figure 2

Sa.LTA-mediated inhibitory effect of B cell proliferation is involved in TLR2. (a) Splenocytes (1 × 10⁶ cells/ml) were isolated from wild-type or TLR2-deficient C57BL/6 mice and stimulated with the indicated concentrations of Sa.LTA and/or LPS for 72 h. Then, cell proliferation was determined by [³H]-thymidine incorporation. Data are expressed as the average cpm ± standard deviation of three separate experiments. *P < 0.05 compared with LPS treatment group without Sa.LTA. (b) CFSE-labeled splenocytes (1 × 10⁶ cells/ml) were incubated with de-acylated LTA, de-alanylated LTA or LTA from the S. aureus Δlgt strain (50 μg/ml each) in the absence or presence of LPS (0.1 μg/ml) for 72 h. Then, the cells were stained with PerCP-conjugated rat anti-mouse CD45R/B220 antibody and B cell proliferation was determined by flow cytometry. NT denotes non-treatment. (c) Splenocytes (1 × 10⁶ cells/ml) were stimulated with Pam2CSK4 and/or LPS at the indicated concentrations for 72 h and cell proliferation was determined by [³H]-thymidine incorporation. Data are expressed as the average cpm ± standard deviation of three separate experiments. *P < 0.05 compared with control culture without LPS stimulation. (d) CFSE-labeled splenocytes were stimulated with Pam2CSK4 (1 μg/ml) and/or LPS (0.1 μg/ml) for 72 h. Then, the cells were stained with PerCP-conjugated rat anti-mouse CD45R/B220 antibody and B cell proliferation was determined by flow cytometry.