FIG 4.

Cytokines produced by PBMCs isolated from pigs in the indicated groups on day 28 (2 weeks after the boost). Pigs in groups 1 and 2 (6 pigs each) were vaccinated with the 5 candidate proteins on days 0 and 14 of the experiment. Both groups were given the 5 proteins intranasally with polyethyleneimine as the adjuvant; in addition, group 1 pigs were given the 5 proteins intramuscularly with AddaVax and Carbopol as the adjuvant, while group 2 pigs were given the 5 proteins intramuscularly with Emulsigen-D as the adjuvant. Pigs in groups 3 to 5 were controls given the adjuvants (groups 3 and 4, 3 pigs each) alone or PBS (group 5, 4 pigs). PBMCs collected on day 28 were stimulated in vitro with a pool of the 5 candidate proteins, and the supernatants were collected to evaluate the levels of cytokines secreted by the cells by multiplex cytokine ELISA. Data are presented as box and dot plots representing the mean cytokine concentration for the group (box) and individual cytokine concentration for each pig (dots) (in picograms per milliliter). Significantly different cytokine concentrations among groups are identified with different letters (P < 0.05).