Active Programmed Cell Death Protein 1 (PD1)

PD-1 (Programmed Death-1 receptor), also known as CD279, is a receptor expressed on T cells responsible for modulating T cell activation. Like CTLA4, PD-1 is classified as an immune checkpoint inhibitory receptor. When PD-1 protein binds to PD-L1, it initiates a negative signaling cascade inhibiting activation of T cells. The cytoplasmic tail contains two tyrosine residues that form the immunoreceptor tyrosine-based inhibitory motif (ITIM) and immunoreceptor tyrosine-based switch motif (ITSM) that are important for mediating PD-1 signaling. Normally, PD-1 helps keep T cells from attacking other cells in the body. However, many cancers take advantage of this by expressing high amounts of PD-L1 allowing cancer cells to evade the body's own immune response. Blocking the PD-1:PD-L1 interaction has proven successful in treating many different cancer types. A functional binding ELISA assay was conducted to detect the interaction of recombinant cat PDCD1 and recombinant rat PDCD1LG2. Briefly, biotin-linked PDCD1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to PDCD1LG2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µl stop solution to the wells and read at 450 nm immediately. The binding activity of PDCD1 and PDCD1LG2 was shown in Figure 1, and this effect was in a dose dependent manner.