Figure 5.

Biochemical effects of aLag-3 and aPD-1 Ab treatment. (A) 2D12 T-cell hybridomas expressing Lag-3 and PD-1 were stimulated with MCC peptide and DC-1 cells with or without addition of 20 nM of control Ab (ctrl), aLag-3 (28G10), aPD-1 (DX400), the cocktail of aLag-3 and aPD-1 Abs, or PMA plus ionophore (Iono+PMA). IL-2 production in the culture supernatant after 24 hours was measured. (B) 2D12 T-cell hybridoma expressing Lag-3 and PD-1 was stimulated with MCC peptide, and DC-1 cells with or without blocking Abs were lysed at the indicated time after stimulation. Tyrosine phosphorylation levels were analyzed by Western blotting using indicated anti-phospho-tyrosine Abs. The quantitation of the blots is shown on the right. * and ** mean p < 0.1 and 0.01, respectively. MCC, moth cytochrome c; Ab, antibody; PMA, phorbol myristate acetate.