Figure 4.
Charge reversal of putative pAg binding pocket residues prevents pAg binding and Vγ9Vδ2 activation. The residues of the B30.2 basic pocket were mutated as follows: H378D, K393D, R412E, R418E, and R469E. A) Shown are the binding isotherms of cHDMAPP with the B30.2 domain (filled squares), the B30.2 charge reversal mutant (filled triangle), and a buffer control (open circles). B) Vγ9Vδ2 T cell stimulation as assessed by CD107a up-regulation is shown in response to no stimulation (No stim.), addition of the α-CD277 agonist antibody 20.1, or in the presence of the bisphosphonate zoledronate (NBP). Data are mean of duplicates ± SD and are representative of three independent experiments. * P <0.05 (paired Student t-test).