In cells stimulated twice by anti-CD3/CD28 antibodies, we found a significant upregulation of activation markers CD69, CD25, CD38 and CD71 (the transferrin receptor) [ 87 ] with simultaneous upregulation of inhibitory markers PD-1, TIGIT and Tim-3 (the latter showed a trend but did not reach statistical significance), which suggested a degree of exhaustion after two rounds of stimulation in a relatively short time interval [ 81 ] ( Fig 9A ).
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Within specific CD4+ T cell populations, CD45RA+ CCR7+ “naïve” cells showed the greatest susceptibility to AUY922, followed by CD45RA- CCR7+ central memory cells [ 86 ] ( Figs 8C and S9 A and S9 B), although the latter trend did not reach statistical significance.