Biocytogen 

Animal Models and Cell Lines

>Single Humanized Immune-Checkpoint Mice

Agonistic CD40 antibodies 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.

Strain description

Strain Name

C57BL/6-Cd40tm1(hCD40)/Bcgen

Background

C57BL/6

Related Genes

Cd40 (CD40   antigen)


Gene Description

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 antibodies 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 antibodies targeting CD40 may serve as a new approach for cancer immunotherapy which is fundamentally different from antagonistic antibodies targeting negative immune checkpoints such as CTLA4 and PD-1.



Targeting strategy

图片.png



Protein expression analysis


图片.png

Fig 1. Splenocytes from both wild type (WT) C57BL/6 and heterozygous B-hCD40 mice were analyzed by flow cytometry. 

Mouse CD40+ B cells were detected in both WT C57BL/6 and heterozygous B-hCD40 mice, while human CD40+ B cells were only detected in the heterozygous B-hCD40 mice.



Human CD40 mAb efficacy evaluation (MC38)


图片.png

Fig 2. Murine colon cancer MC38 cells were subcutaneously implanted into heterozygous B-hCD40 mice. Mice were grouped when the tumor size was approximately 150±50mm3 (n=5). Three human CD40 antibodies differently inhibited tumor growth, confirming that the B-hCD40 mouse model is a powerful tool for in vivo CD40 antibody pharmacological efficacy studies. (A) Tumor average volume ± SEM, (B) Mice average weight ± SEM.


图片.png

Fig 3. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hCD40 mice. Mice were grouped when the tumor size was approximately 150±50mm3 (n=5). Selicrelumab Analog (human CD40 antibody) significantly inhibited tumor growth, confirming that the B-hCD40 mouse model is a powerful tool for in vivo CD40 antibody pharmacological efficacy studies. (A) Tumor average volume ± SEM, (B) Mice average weight ± SEM.


Human CD40 mAb toxicity evaluation


图片.png

Fig 4. Homozygous B-hCD40 mice were i.p. injected with vehicle control and different Anti-human CD40 antibodies on days indicated by red arrows (n=5).

No significant difference in average body weight was detected after the treatments. 


图片.png

Fig 5. The blood routine examination was performed with anti-CD40 antibody on B-hCD40 mice two days after the termination of efficacy evaluation experiment.


Abbreviation

Stand for

Abbreviation

Stand for

WBC

White Blood Cell Count

MCV

Mean Corpuscle (Cell) Volume  

NE#

Neutrophil Count

MCH

Mean Corpuscular Hemoglobin

LY#

Lymphocyte Count

MCHC

Mean Corpuscular Hemoglobin Concentration

MO#

Monocyte Count  

MPV

Mean Platelet Volume

RBC

Red Blood Cell Count

PLT

Platelet Count  

Hb

Hemoglobin

RDW

Red Cell Distribution Width

HCT

Hematocrit  


图片.png


Fig 6. The blood biochenmical index was examined with anti-human CD40 antibody on B-hCD40 mice two days after the termination of efficacy evaluation experiment.


Abbreviation


Stands for  

Explanation

ALT

Alanine transaminase

High level indicates liver injured  

AST

Aspartate

transaminase

High level indicates liver damage  

CHOL

Cholesterol

High level   indicates high blood lipid  

CR

Creatinine

High level   indicates low glomerular filtration  

GLU


Glucose  

Hyperglycemia or hypoglycemia


TRIG  

Triacylglycerol

High level   indicates high blood lipid

UREA


Urea    

High level indicates kidney damage, liver disease, diabetes or infection.  


图片.png

Fig 7. Flow cytometry was performed to examine lymphocytes lineage after anti-human CD40 antibody treatment on B-hCD40 mice. The results indicated that CD40 Ab4 treatment resulted in a significant increase in the percentage of T cells and B cells, respectively.


图片.png

Fig 8. Concentrations of different cytokines were measured after anti-human CD40 antibody treatment on B-hCD40 mice. The results indicated that CD40 Ab4 treatment resulted in a significant increase in the concentration of several cytokines, such as IFN-γ, TNF-α MCP-1 and IL 27.  



Activation of B lymphocyte in B-hCD40 mice


图片.png

Fig 9. Activation of B lymphocyte in B-hCD40 mice. Homozyous B-hCD40 mice were i.p. injected with vehicle control and different Anti-human CD40 antibodies. Afther 48h, Abdominal cells and splencytes were collected and analyzed the expression activation markers by flow cytometry.




【Return list】
* Contact Name
* Quantity
Telephone No.
Email
* Shipping address
Verification code captcha

© 2018 Beijing Biocytogen Co., Ltd. All rights reserved. Technical support: fast help cloud
in the section of your HTML pages