Figure 3
HSV-specific proliferation of splenocytes and LN cells obtained from pCIgB-immunized mice and cAMP production by salbutamol. (A) Proliferation of splenocytes and LN cells after specific restimulation with HSV antigen. Groups of BALB/c (H-2d) mice were co-immunized i.n. with a mixture of pCIgB DNA vaccine (100 µg) and salbutamol (10, 50, 100, and 200 µg) three times at 7-day intervals. Two weeks following the final immunization, the responder cells were prepared from spleen and draining LN, and mixed with irradiated syngeneic-enriched APCs that had been pulsed with UV-inactivated HSV. (B) Increased HSV-specific proliferation of CD4+ T cells in the presence of salbutamol. CD4+ T cells purified from HSV-immunized mice were mixed with HSV antigen-pulsed APCs in the presence of the indicated salbutamol concentrations. Following 3-day incubation, the proliferated cells were determined by [H3] thymidine incorporation. The graph represents the average ± SD of values obtained from three or four individual experiments. (C) The production of cAMP in dendritic cells treated with salbutamol. DC2.4 cells were treated with the indicated concentration of salbutamol in presence or absence of propranolol (β-adrenergic antagonist) and used for determination of intracellular cAMP levels. CT (cholera toxin) was included for positive control. The bars represent the average ± SD of values obtained from quadruplicate wells. ***P < 0.001 when compared between the indicated groups.
