Fig. 3. Both CD4⁺ and CD8⁺ subsets of TFA-specific T cells were generated in BALB/cByJ mice immunized by TFA-MSA and the combined CD40/CpG agonist.

Female BALB/cByJ mice were injected i.p. with 0.5 mg of TFA-MSA, 40 µg of anti-CD40 Ab, and 100 µg of CpG. One week later, mice were sacrificed and splenocytes isolated. CD4⁺ or CD8⁺ T cells were further purified by using a T cell enrichment kit. Subsequently, 2.5× 10⁶ of CD4⁺ T cells (A) and 1.25× 10⁶ of CD8⁺ T cells(B) per well of 48-well plate were cultured with naïve splenocytes (1 × 10⁶ cells/well), which served as APCs, in the presence of various antigens (50 µg /ml) for 48 h. IFN-γ production was measured in the supernatant by ELISA. The measurements were carried out in triplicate and the results shown represent mean ± SEM of 3 mice per group. *P<0.05 compared with the LMNC stimulated with CCH. The data shown is the representative of two independent experiments.