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. 2012 Nov 27;21(3):542–553. doi: 10.1038/mt.2012.233

Figure 5.

Figure 5

Major histocompatibility complex class I presentation of ovalbumin (OVA) peptide to OVA-specific CD8+ T cells by TC-1/Meso tumor cells treated with Meso-scFv-ROR-Fc. (a) Representative flow analysis of Meso-scFv-ROR-Fc protein binding to TC-1/Meso. The purified chimeric proteins were incubated with TC-1/Meso or control TC-1 tumor cells followed by staining with phycoerythrin-labeled antimouse Fc antibody. (b) Flow cytometry characterization of OVA-specific CD8+ T-cell activation by TC-1/Meso cells treated with different chimeric proteins. OVA-specific CD8+ T-cell activation was determined by CD8 and intracellular IFN-γ staining. (c) Representative bar graph depicting the % of IFN-γ-secreting OVA-specific CD8+ T cell out of total OVA-specific CD8+ T cells (mean ± SD). (d) Representative luminescence imaging of in vitro OVA-specific cytotoxic CD8+ T lymphocytes (CTL) killing of luciferase-expressing TC-1/Meso cells treated with Meso-scFv-ROR-Fc. Degree of CTL-mediated tumor cell death is indicated by decrease of luminescence activity. (e) Bar graph depicting viability of tumor cells treated with protein and/or OVA-specific cytotoxic T cells (mean ± SD) (representative data of two experiments). Meso-scFv, mesothelin single-chain variable fragment.