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. Author manuscript; available in PMC: 2019 Jul 1.
Published in final edited form as: Cancer Immunol Res. 2018 Sep 21;6(12):1511–1523. doi: 10.1158/2326-6066.CIR-18-0193

Figure 1: OT-I CTL screen design.

Figure 1:

(A) ID8-Cas9 serous ovarian carcinoma cell line was transduced with a pLVX vector expressing either firefly luciferase fused to a model antigen peptide or renilla luciferase and no antigen. Clonal cell lines were generated using G418 selection for Neo cassette expression and limiting dilution. (B) 10,000 ID8-lucOS and 10,000 ID8-rluc were co-plated into wells of 96-well tissue culture plates. OT-I TCR transgenic CD8+ T cells were then plated on top of ID8 cells. Total volume/well was 200 μL and was incubated for 48hrs. at 37°C and 5% CO2 prior to analysis by dual-luciferase assay. (C) OT-I assay was screened in high-throughput to evaluate compounds for immunomodulatory effects on antigen-specific tumor cell killing by cytotoxic T lymphocytes (CTLs). Inclusion of ID8-lucOS and ID8-rluc provided in-plate normalization controls to determine non-specific growth inhibition or induction of apoptosis by screen compounds.