Extended Data Fig. 9: PD-1 and GITR receptor copy number, expression and anti-PD-1-GITR-L cross-reactivity on normal human and cynomolgus monkey tissues by IHC, and Hexagonal conformation model of GITR-L-GITR complex.
(A and B) GITR and PD-1 receptor copy number on PHA activated (t = 48 hrs) human (A) and cynomolgus monkey (B) PBMCs (n = 8 donors per group). Data are presented as mean values +/- SEM. Statistical significance was calculated with two-way ANOVA with Tukey’s correction for multiple comparisons (statistic refer to GITR vs PD-1 expression). (C) Representative human and Cyno PD-1 and GITR expression by IHC (representative staining of n = 2 tissues) on tonsil, lymph node, spleen, and stomach tissues. FFPE tissue samples were used for PD-1 and GITR (Human TMA: ZPL2 and ZPL3, and Cyno: I09447 and I08762) staining. Scale bars indicate 50 µm (PD-1) and 60 µm (GITR) (original magnification 40x). (D) Representative anti-huPD-1-huGITR-L bispecific binding on normal human and cynomolgus monkey tissues using human to human pre-complex conjugated IHC staining method. Scale bars indicate 200 µm (original magnification 20x, representative staining of n = 2 tissues). (E) GITR-L trimeric units are shown in light blue, and GITR monomeric units from different GITR trimers are shown in pink and orange.
