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. 2019 Mar 11;1:100017. doi: 10.1016/j.jvacx.2019.100017

Fig. 4.

Fig. 4

KLIpS immunization could induce specific cytolytic activity. (A) AAD transgenic mice (n = 5) were immunized twice with KLIpS (triangle), KLIS (square) or PBS (control, circle) formulated with PADRE in IFA. Severn days after immunization, the immunized mice were injected with CFSE-labeled peptide-pulsed spleen cells to exam the in vivo cytolytic activity of CTLs. The specific lysis percentage represented the reduced amount of peptide-pulsed spleen cells after 18 hrs post-adoptive transfer. (Student’s t-test, KLIS vs KLIpS, **, p < 0.01). (B) 5 × 105 CD8+ T cells were purified from spleens of KLIpS-immunized transgenic mice and co-cultured with 5 × 103 51Cr-labeled H2981 for 18 h. The 51Cr-release assay result shows that KLIpS-stimulated mouse CD8+ T cells could cytolysis human lung H2981 cancer cell. 51Cr-labeled H2981 cells were incubated with 20 μg/ml anti-HLA Ab (W6/32), or purified CD8+ T cells were incubated with 10 μg/ml anti-CD8 Ab (53–6.7) for 15 min before co-culture. Addition of specific antibody could significantly inhibit the cytolysis activity of mouse CD8+ T cell. (w/o Ab vs anti-HLA Ab, *, p < 0.05; w/o Ab vs anti-CD8 Ab, **, p < 0.01). Specific lysis percentage was calculated using formula: 100 × [(experimental release − spontaneous release)/(maximal release - spontaneous release)].