Description 

Gene targeting strategy for B-hCD112 MC38 cells. The exogenous promoter and chimeric CD112 containing mouse signal peptide, human extracellular region and mouse transmembrane and intracellular region was inserted to replace part of murine exon 2 and all of exon 3. The insertion disrupts the endogenous murine Cd112 gene, resulting in a non-functional transcript.

Protein expression analysis 

Subcutaneous homograft tumor growth of B-hCD112 MC38 cells. B-hCD112 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 8-week-old). 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-hCD112 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hCD112 MC38 cells. B-hCD112 MC38 cells (5x10⁵, 3x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into B-hPVRIG mice (female, 9-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-hCD112 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

B-hCD112 MC38 cells were subcutaneously transplanted into B-hPVRIG mice, and on 35 days post inoculation, tumor cells were harvested and assessed for human CD112 expression by flow cytometry. As shown, human CD112 was highly expressed on the surface of tumor cells. Therefore, B-hCD112 MC38 cells can be used for in vivo efficacy studies of novel CD112 therapeutics.