Figure 4.
Proliferative response of lymphocytes belonging to various immunized groups upon stimulation with SAgs. To determine the effect of the amount of SAgs on the proliferation of lymphocytes, lymphocytes isolated from the spleens of immunized BALB/c mice of various vaccinated groups including controls, were cultured in flat-bottomed 96-well plates. The cells were co-cultured in the presence of increasing doses (1–20 μg) of corresponding formulations of SAgs as well as controls. After 72 hours [3H]-thymidine was added and its incorporation into multiplying cells was measured after 16 hours of incubation, using liquid scintillation spectroscopy. The CPM values of stimulated culture were used to represent Ag-specific stimulation. (A) Dose-dependent proliferative response of lymphocytes at 1-week post-booster and post-challenge (data not shown) upon stimulation with various SAgs formulations. (B) Lymphocyte proliferation in terms of CPM values in various immunized groups at PB and 2 weeks PC time points.
Notes: Data represent the mean of three determinants ± SD. The figures are representative of three independent experiments. The groups were compared using ANOVA. Holm–Sidak test (all pairwise multiple comparison procedures) with *** P < 0.001.
Abbreviations: CPM, counting per minute; PB, post-booster; PC, post-challenge; PBS, phosphate buffer saline; ANOVA, analysis of variance; SD, standard deviation; SAgs, secretory protein antigens.