B-hPD-L1 B16-F10_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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B-hPD-L1 B16-F10

Common name	B-hPD-L1 B16-F10	Catalog number	310687
Aliases	CD274, PDCD1L1	Disease	Melanoma
Organism	Mouse	Strain	C57BL/6
Tissue types	Skin	Tissue	Skin

Description

The mouse Pdl1 gene was replaced by human PD-L1 coding sequence in B-hPD-L1 B16-F10 cells. Human PD-L1 is highly expressed on the surface of B-hPD-L1 B16-F10 cells.

Application

B-hPD-L1 B16-F10 cells have the capability to establish tumors in vivo and can be used for efficacy studies.

Targeting strategy

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 B16-F10 cells by flow cytometry. Single cell suspensions from wild-type B16-F10 and B-hPD-L1 B16-F10 cultures were stained with species-specific anti-PD-L1 antibody. Human PD-L1 was detected on the surface of B-hPD-L1 B16-F10 cells but not wild-type B16-F10 cells. The 2-D3 clone of B-hPD-L1 B16-F10 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

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Subcutaneous homograft tumor growth of B-hPD-L1 B16-F10 cells. Wild-type B16-F10 and B-hPD-L1 B16-F10 cells (1x10⁵) 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-L1 B16-F10 cells were able to establish tumors in vivo and can be used for efficacy studies.

PD-1 Ab (Keytruda) combined with OX40 Ab efficacy evaluation

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The B-hPD-L1 B16-F10 cell line was subcutaneously implanted into homozygous B-hPD-1/hOX40 mice. Mice were grouped when tumor size reached approximately 150 ± 50 mm³ (n = 5). (A) Mean tumor volume ± SEM; (B) Mean body weight of mice ± SEM.

The result shows a moderate antitumor activity by the Keytruda or OX40 Ab compared with the vehicle control group. The data also indicated that Keytruda combined with OX40 Ab potentiated the antitumor activity of the Keytruda or OX40 Ab alone.