Identification of CTL Epitopes on Efflux Pumps of the ATP-Binding Cassette and the Major Facilitator Superfamily of Mycobacterium tuberculosis

Figure 4

Activity of CTLs induced with IFA-emulsified peptide in HLA-A2.1/K^b transgenic mice. HLA-A2.1/K^b transgenic mice () were subcutaneously immunized at the base of the tail with each peptide (100 μg/mice) and T helper epitope (140 μg/mice) emulsified with incomplete Freund’s adjuvant (IFA) at 1 : 1, three times (on days 0, 5, and 10). On day 11, all the mice were sacrificed and the serum was collected from each mouse to measure the IFN-γ concentration with an ELISA assay (a). The splenic lymphocytes were isolated from each mouse and restimulated once with each peptide (10 μg/mL) and mIL-2 (10 U/mL) in vitro for another 6 days. On day 7, the IFN-γ release in each group was measured with an intracellular cytokine staining (ICS) assay (b) and the lysis activity was measured with an LDH assay (c). Peptide-loaded T2A2 cells were used as the target cells. Normal saline (NS) group and Th epitope only group were the negative controls. Statistical significance was determined with Student’s -test (). and represent the significance of differences relative to the NS group.

(a)

(b)

(c)