The mouse PD-L2 gene was replaced by human PD-L2 coding sequence in B-hPD-L2 MC38 cells. Human PD-L2 is highly expressed on the surface of B-hPD-L2 MC38 cells.

PD-L2 expression analysis in B-hPD-L2 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hPD-L2 MC38 cultures were stained with species-specific anti-PD-L2 antibody. Mouse PD-L2 was not detected on the surface of wild-type MC38 cells. Human PD-L2 was detected on the surface of B-hPD-L2 MC38 cells but not wild-type MC38 cells. The 1-C11 clone of B-hPD-L2 MC38 cells was used for in vivo experiments.

Subcutaneous homograft tumor growth of B-hPD-L2 MC38 cells. B-hPD-L2 MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 5-8-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean± SEM). Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hPD-L2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hPD-L2 MC38 cells. B-hPD-L2 MC38 cells (5x10⁵, 1x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into B-hCD3E mice (female, 7-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean± SEM). Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hPD-L2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.