Gene targeting strategy for B-hCD40/hCD40L mice(C). 

The exons 2-7 of mouse Cd40 gene that encodes the extracellular region were replaced by human CD40 exons 2-7 in B-hCD40/hCD40L mice(C). 

The exons 2-5 of mouse Cd40l gene that  encode the extracellular region were replaced by human CD40L exons 2-5 in B-hCD40/hCD40L mice(C).

B-hCD40/hCD40L mice(C) were used to establish T cell-Dependent Antibody Response assay (TDAR assay) and evaluate the efficacy of anti-CD40 antibody and anti-CD40L antibody. B-hCD40/hCD40L mice(C) (n=6) were intraperitoneally immunized with 200 μg KLH on day 1 and treated with anti-CD40L antibody dapirolizumab analog (in house) or anti-CD40 antibody bleselumab analog (in house) on day 0 and day 4. Blood was collected on day 7 and day 14 and analyzed by ELISA with KLH specific IgG antibody and KLH specific IgM antibody. (A) Body weight of B-hCD40/hCD40L mice(C) increased steadily; (B, C) Concentration of mouse KLH specific IgM and IgG were significantly increased after immunization. But the concentration of KLH specific IgG and IgM in the groups treated with anti-CD40L antibody dapirolizumab analog (in house) or anti-CD40 antibody bleselumab analog (in house) were significantly decreased when compared to that in the control group. The concentration of KLH specific IgG in the groups treated with anti-CD40L antibody dapirolizumab analog (in house) was not decreased on day 14, which may be due to incomplete blockade of B cells by inhibiting the CD40-CD40L pathway. These results demonstrated that the B-hCD40/hCD40L mice(C) provide a powerful preclinical model for in vivo evaluation of anti-CD40 antibody and anti-CD40L antibody. Values are expressed as mean ± SEM. Significance was determined by one-way ANOVA test . * p < 0.5, **p < 0.01, ***p < 0.001.