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. 2015 Jul 29;6(28):25356–25367. doi: 10.18632/oncotarget.4516

Figure 4. Induction of human CXorf61-specific T cells in vitro and cloning of a CXorf61-specific human TCR.

Figure 4

A. CD8+ T cells of a healthy HLA-A*0201 expressing donor were primed using autologous mDC transfected with CXorf61 RNA. After three rounds of stimulation CXorf61-specific T cells were detected by CXorf6166–74/A*0201 dextramer staining and isolated by flow cytometry for TCR cloning. B. Specificity testing of a TCR isolated from human CD8+ T cells sensitized against CXorf61. CD8+ T cells transfected with the cloned TCR chains were tested by IFNγ-ELISPOT for recognition of K562-A*0201 cells either transfected with CXorf61 IVT RNA or loaded with CXorf6166–74; negative controls: irrelevant RNA CLDN6 (control RNA); irrelevant peptide PLAC131–39 (control peptide); positive control: SEB treated cells.