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. 2018 Apr 19;115(19):E4453–E4462. doi: 10.1073/pnas.1800550115

Fig. 3.

Fig. 3.

Induction of hTERT-specific CTLs by ihv-DCs in an autologous setting. (A) The hTERT (amino acids 301 to 700) fragment that covers six HLA-A2–restricted epitopes is depicted. (B) Detection of the proteasome-targeted TERT (amino acids 301 to 700) or pTERT in ihv-DC2-pTERT cells or ihv-DC2-pTERT-Allo cells that were pretreated with DMSO or MG132 for 1 h. ihv-DC2-pTERT-Allo cells were generated by lentiviral transduction of DRB1*0701/DRB1*1301. (C) Immunophenotype of ihv-DC2-pTERT–primed, CD3/CD8-enriched T cells. (D) Immunophenotype of ihv-DC2-pTERT-Allo–primed, CD3/CD8-enriched T cells. (E) Cytokine expression profiles of the ihv-DC2-pTERT-Allo–primed, CD3/CD8-enriched T cells as examined by qRT-PCT. (F) Anti-hTERT immunoblot analysis for A375, A549, and U2OS cancer cells. (G) Fluorescence imaging analysis of U2OS cells transduced with FLAG-TERT(aa301–700) using anti-FLAG antibody. Luc, luciferase. (H) Cytotoxicity assay using U2OS-Luc or U2OS-Luc/TERT cells as target cells, with ihv-DC2-pTERT–primed, CD3/CD8-enriched T cells as killer cells. (I) Cytotoxicity assay using U2OS-Luc or U2OS-Luc/TERT cells as target cells, with ihv-DC2-pTERT-Allo–primed, CD3/CD8-enriched T cells as killer cells. (J) Cytotoxicity assay using U2OS-Luc or U2OS-Luc/TERT cells as target cells, with ihv-DC2-pTERT-Allo–primed, CD3/CD8-enriched T cells as killer cells, in the presence of control antibody or anti-HLA-A2 antibody (30 μg/mL). (K) Cytotoxicity assay using the melanoma cells A375 as target cells, with ihv-DC2-pTERT-Allo–primed, CD3/CD8-enriched T cells as killer cells, in the presence of increasing amounts of the control antibody or anti-HLA-A2 antibody. (L) Cytotoxicity assay using A549-Luc or A549-Luc/A2.1 cells as target cells, with ihv-DC2-pTERT-Allo–primed, CD3/CD8-enriched T cells as killer cells. *P < 0.05, **P < 0.01, as determined by two-tailed Student’s t test.