Fig. 4.

MET inhibitors suppress the immunosuppressive capabilities of recombinant HGF. A, B Peripheral blood mononuclear cells (PBMCs) activated with phytohemagglutinin (PHA, 10 µg/ml) were cultured either alone (A) or in the presence of hepatocyte growth factor (HGF, 75 ng/mL) and/or MET inhibitors (crizotinib, SGX-523). The MTS proliferation assay was employed to quantify lymphocyte/PBMC proliferation at 72 h, while cell supernatants were collected at 24 h for interferon-gamma (IFNγ) quantification. C, D Activated PBMCs were stained with CFSE and treated for 96 h with HGF (75 ng/mL) alone or in combination with pembrolizumab (20 nm) and/or MET inhibitors (crizotinib, SGX-523). The stained cells were collected, probed with PE-Cy7-conjugated anti-CD3, BV421-conjugated anti-CD45, and APC-conjugated anti-CD8 antibodies, and analyzed using flow cytometry. Data are presented as mean ± SEM of at least four independent experiments. *p < 0.05 and ***p < 0.001