Figure 2. Engineered murine TCR₁₀₄₅ T cells lyse murine ovarian cancer cells.

A) High-resolution ultrasound of ID8_VEGF tumor development in the peritoneal cavity (8 weeks after injection). White arrows with circles demarcate tumor nodules in the pancreas.

B) High resolution ultrasound of advanced disease. Anechoic ultrasound pattern indicates ascites development in the peritoneal cavity and obscures tissue landmarks.

C) Immunohistochemistry for MSLN/Msln in human high-grade serous ovarian cancer or ID8_VEGF tumors. Images are representative of 15 ID8_VEGF tumors and 31 HGSOC samples.

D) Immunoblot analyses of wild type or Msln knockout mouse tissues and ID8_VEGF tumors for Msln protein expression. Vinculin was used as the loading control. Representative of 2 independent experiments.

E) MHC I expression on ID8_VEGF tumor cells with or without IFNγ exposure (10ng/ml for 24 hours).

F) CFSE dilution of TCR₁₀₄₅ or control TCR_OTI T cells after co-culture with ID8_VEGF tumor cells for 5 days.

G) Cleaved Caspase 3 expression in ID8_VEGF tumor cells after co-culture with TCR₁₀₄₅ or control TCR_OTI T cells for 16 hours. Dotted line at 0.64% represents average frequency of CC3⁺ staining in ID8_VEGF tumor cells at the equivalent time after plating without co-culture.

H) Cytolysis of ID8_VEGF tumor cells after co-culture with TCR₁₀₄₅ or irrelevant TCR_OTI T cells for 16 hours, measured by impedance of an electrical signal by target cells (xCELLigence platform). Dotted line at 1.89% represents the average background lysis by TCR_OTI T cells.

I) Percent of ID8_VEGF target cell death by live cell quantification after co-culture with TCR₁₀₄₅ or control TCR_OTI T cells for 16 hours. All data are representative of 2–3 independent experiments. White arrows indicate tumor nodules. Scale bar = 50 microns. Data are shown +/− SD.