B-hPD-L1 LLC1

Basic Information

Description

The mouse Pdl1 gene was replaced by human PD-L1 coding sequence in B-hPD-L1 LLC1 cells. Human PD-L1 is highly expressed on the surface of B-hPD-L1 LLC1 cells.

Targeting strategy

Gene targeting strategy for B-hPD-L1 LLC1 cells. The exogenous promoter and human PD-L1 coding sequence was inserted to replace part of murine exon 3. The insertion disrupts the endogenous murine Pdl1 gene, resulting in a non-functional transcript.

Protein expression analysis

PD-L1 expression analysis in B-hPD-L1 LLC1 cells by flow cytometry. Single cell suspensions from wild-type LLC1 and B-hPD-L1 LLC1 cultures were stained with species-specific anti-PD-L1 antibody. Mouse PD-L1 was detected on the surface of wild-type LLC1 cells. Human PD-L1 was detected on the surface of B-hPD-L1 LLC1 cells but not wild-type LLC1 cells. The 1-A5 clone of B-hPD-L1 LLC1 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hPD-L1 LLC1 cells. B-hPD-L1 LLC1 cells (2×10⁵) were subcutaneously implanted into C57BL/6 mice (female, 7-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-L1 LLC1 cells were able to establish tumors in vivo and can be used for efficacy studies.

Protein expression analysis of tumor cells

B-hPD-L1 LLC1 cells were subcutaneously transplanted into C57BL/6 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human PD-L1 expression by flow cytometry. As shown, human PD-L1 was highly expressed on the surface of tumor cells. Therefore, B-hPD-L1 LLC1 cells can be used for in vivo efficacy studies of novel PD-L1 therapeutics.