Figure 9.

Analysis of cellular memory development. The analysis of proliferation of mouse spleen lymphocytes was performed in the presence of recombinant proteins (as described in Materials and Methods). (a) Spleen lymphocytes of threefold immunized mice incubated in vitro with the same protein that was used for vaccination. Spleen lymphocytes of the adjuvant control (b) and untreated (c) mice were incubated with recombinant proteins without earlier contact with them. In all groups, PHA (5 μg/ml) was applied as a positive control. Each bar presents % of the viability of control cells (lymphocytes of nonimmunized mice) from six individual lymphocyte suspensions performed in duplicate. The results are expressed as the mean ± S.E.M. The differences were analyzed statistically with the Kruskal-Wallis test followed by the Dunn multiple comparison test. (a) ^∗Statistically significant differences in comparison to the control cells (white bars) of immunized mice, ^∗P ≤ 0.05. ⁺Significantly different in comparison to the lower protein concentration, P ≤ 0.05. ^#Significantly different in comparison to the corresponding results obtained in the adjuvant control from (b), ^#P ≤ 0.05. (b, c) ^∗Differences are statistically significant in comparison to the protein-untreated cells of the control (untreated) animals. The viability of these cells was estimated at 100%. ⁺Significantly different in comparison to the lower protein concentration, ^∗P ≤ 0.05.