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B-hPD-L1/hCD40 mice
Strain Name C57BL/6-Cd274tm1(CD274)Cd40tm1(CD40)/Bcgen Common Name  B-hPD-L1/hCD40 mice
Background C57BL/6 Catalog number  120536
Related Genes 
CD274 (CD274 antigen)
CD40 (CD40 antigen)

Gene description

PD-L1 (Programmed cell death ligand-1), also known as B7-H1 and CD274, is mainly expressed in antigen-presenting cells (APCs) and activated T cells, and is one of the two ligands of PD-1. The interaction between PD1 and PD-L1 plays an important role in the negative regulation of the immune response. PD-L1 is highly expressed in a variety of solid tumors. PD-1 and PD-L1 interactions can reduce T Cell Activation and promote tumor immune escape. The PD-1/PD-L1 signaling pathway can be blocked and antitumor immune response can be restored by using by anti-PD-1 or anti-PD-L1 antibodies to block the binding of PD1 to PD-L1. CD40 (cluster of differentiation 40) is a tumor necrosis factor receptor superfamily member expressed on APC such as dendritic cells (DC), B cells, and monocytes as well as many non-immune cells and a wide range of tumors. Interaction with its trimeric ligand CD154 on activated T helper cells results in APC activation, which has been found to be essential in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. Agonistic CD40 mAb have been shown to activate APC and promote anti-tumor T cell responses and to foster cytotoxic myeloid cells with the potential to control cancer in the absence of T-cell immunity. Thus, agonistic CD40 mAb may serves as a new antitumor drugs which are fundamentally different from mAb which block negative immune checkpoint such as anti-CTLA-4 or anti-PD-1.


Targeting strategy


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Gene targeting strategy for B-hPD-L1/hCD40 mice. 

The exon 3 of mouse Pd-l1 gene that  encodes the extracellular domain was replaced by human PD-L1 exon 3 and  the exons 2-7 of mouse Cd40 gene that  encode the extracellular domain were replaced by human CD40 exons 2-7 in B-hPD-L1/hCD40 mice. 


Protein expression analysis


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Strain specific CD40 and PD-L1 expression analysis in homozygous B-hPD-L1/hCD40 mice by flow cytometry. 

Splenocytes were collected from WT and homozygous B-hPD-L1/hCD40 (H/H) mice analyzed by flow cytometry with species-specific anti-PD-L1 antibody and anti-CD40 antibody. Mouse CD40 and PD-L1 were detected in WT. Human CD40 and PD-L1 were exclusively detected in H/H B-hPD-L1/hCD40 but not WT mice.


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Strain specific CD40 and PD-L1 expression analysis in homozygous B-hPD-L1/hCD40 mice by flow cytometry. 

Splenocytes were collected from WT and homozygous B-hPD-L1/hCD40 (H/H) mice stimulated with anti-CD3ε in vivo (7.5 μg/mice), and analyzed by flow cytometry with species-specific anti-PD-L1 antibody. Mouse CD40 and PD-L1 were detected in WT. Human CD40 and PD-L1 were exclusively detected in H/H B-hPD-L1/hCD40 but not WT mice.


Combination therapy of PD-L1 mAb and CD40 mAb


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Antitumor activity of anti-hCD40 antibody combined with anti-hPD-L1 antibody in B-hPD-L1/hCD40 mice. 

(A) Anti-hCD40 antibody combined with anti-hPD-L1 antibodies inhibited MC38-hPD-L1 tumor growth in B-hPD-L1/hCD40 mice. Murine colon cancer MC38-hPD-L1 cells (5×105) were subcutaneously implanted into homozygous B-hPD-L1/hCD40 mice (female, 6-8 week-old, n=5). Mice were grouped when tumor volume reached approximately 80-120 mm3, at which time they were treated with Anti-hCD40 antibody combined with anti-hPD-L1 antibody and schedules indicated in panel. (B) Body weight changes during treatment. As shown in panel A, combination of anti-hCD40 antibody and anti-hPD-L1 antibody shows more inhibitory effects than individual groups, demonstrating that the B-hPD-L1/hCD40 mice provide a powerful preclinical model for in vivo evaluating combination therapy efficacy of hCD40 antibodies and hPD-L1 antibodies . Statistical results showed significant weight loss in both G2 and G4 groups. As shown in panel B , anti-CD40 antibody significantly reduced body weight ,combination with PD-L1 antibody could relieve this symptom well.Values are expressed as mean ± SEM.