Detection of intracellular cytokine production of splenocytes in mice by flowcytometry after cercarial challenge. Spleen cells of mice (three mice each group) immunized with the SjGP-3 formulated by various adjuvants were harvested on week 12 (three weeks after cercarial challenge) and 15 (six weeks after cercarial challenge). The isolated splenocytes from individual mice were first stimulated with PMA, ionomycin plus BFA. Splenocytes were stained extracellularly by PE-Cy5 labeled anti-CD3 and FITC labeled anti-CD8. After fixation and permeabilization, the spleen cells were divided into three tubes and stained intracellularly with PE conjugated Rat IgG1, anti-mouse IFN-γ antibody or anti-mouse IL-4 antibody, respectively. A FACScan flow cytometer with Cell Quest software was used for data acquisition and analysis. (A) Week 12. (B) Week 15. * p < 0.05 when compared with PBS control group.