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Fig. 1 | Journal for ImmunoTherapy of Cancer

Fig. 1

From: Split tolerance permits safe Ad5-GUCY2C-PADRE vaccine-induced T-cell responses in colon cancer patients

Fig. 1

Ad5-GUCY2C-PADRE design and antigen expression. a GUCY2C is a membrane-spanning enzyme possessing an extracellular ligand-binding domain and intracellular cGMP-producing catalytic domain. The extracellular domain (ECD) of GUCY2C was employed in the vaccine design and included the PADRE epitope on its COOH-terminus. b GUCY2CECD-PADRE was inserted into the E1 region of E1/E3-deleted Ad5. c A549 cells were transduced in duplicate with Ad5-GUCY2C-PADRE at an MOI of 10 to 10,000 for 48 h. GUCY2CECD-PADRE expression was quantified in supernatants by immunoblot analysis. Densitometry was employed to quantify expression. d A549 cells were transduced in duplicate with Ad5-GUCY2C-PADRE at an MOI of 10,000 for 24–96 h and GUCY2CECD-PADRE expression was quantified in supernatants by immunoblot analysis. Densitometry (arbitrary units) was employed to quantify expression. Blots in c and d are representative of two experiments and graphs indicate the mean ± SD from 2 experiments

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