Fig. 8.
Isoproterenol and T cell activation stimulated the release of ACh by isolated splenocytes from male ChATBAC-eGFP mice. Isolated splenocytes were incubated in 2 ml of medium for indicated times with no treatment, 1 μM (−) isoproterenol, or T cell activating antibodies (anti-CD3 and anti-CD28). For each experiment, data were evaluated by repeated measures ANOVA followed by post hoc comparison of group data. A: Isoproterenol increased ACh release after 4 and 24 h incubation, with 24 h causing significantly higher ACh concentration in the medium. B: Addition of T cell antibodies to the medium caused a significant increase in ACh release only after 24 h. C: Treatment with isoproterenol + T cell activation caused greater ACh release than either treatment alone. For T cell activation in this experiment, plates were coated with anti-CD3 overnight, washed before adding cells, and treated with anti-CD28 added to the medium. Data in each panel were evaluated by repeated measures one-way ANOVA, and post hoc comparisons were made using the Tukey test when overall group differences were detected. Values are means ± SEM with n = 7 in A, n = 3 in B and n = 5 in C. *P < 0.05 vs indicated groups.