Melanoma Cells Treated with GGTI and IFN-γ Allow Murine Vaccination and Enhance Cytotoxic Response against Human Melanoma Cells
Figure 4
On LB1319-MEL cells, GGTI-298 enhances hIFN-γ-induced TAP1 and TAP2 expressions.
LB1319-MEL cells were cultivated for 4 days in the presence or in the absence of hIFN-γ (50 IU/ml) and/or GGTI-298 (10 µM) as indicated. A) The expression of molecules implicated in the MHC-I Ag processing pathway were tested in these treated tumor cells. β-actin was used as protein loaded control. The expressions were quantified by calculating the ratio between the protein of interest and the β-actin. We defined the ratio of relevant protein over β-actin for untreated cells equal to 1. B) Expressions of TAP1 and TAP2 proteins in these in vitro treated and permeabilized LB1319-MEL were tested by flow cytometry using TAP1 and TAP2 specific mAbs and a PE-conjugated secondary Ab. C) TAP1 membrane expression was also tested by cytofluorometry on LB1319-MEL cells after 4 days treatment with either increasing doses of hIFN-γ (0, 25, 50 and 100 IU/ml) or with the combination of hIFN-γ (50 IU/ml) and GGTI-298 (10 µM). Results are expressed in ISF, related to isotype controls, as indicated in Material and Methods. Data illustrated are representative of 3 independent experiments.