Fig 1. Detection of CD40 expression in peripheral blood cells of humanized CD40 mice. The FACS results of peripheral blood cells collected from homozygous humanized CD40 mice and wild-type mice showed that the active expression of humanized CD40 was detected in CD19 positive cells collected from homozygous humanized CD40 mice, and its expression level was similar to that of murine Cd40 expression in wild-type mice.(Completed in collaboration with CrownBio).

Fig 2. Analysis of human CD40 expression in the spleen by FACS. The homozygous KI animal expresses human CD40 on the CD19⁺ B cells.

Fig 3. Representative flow cytometry staining of GC B cells frommesenteric lymph nodes.

Fig 4. In vivo validation of anti-tumor efficacy in a MC38 tumor-bearing model of humanized CD40 mice. Humanized CD40 mice were inoculated with MC38 colon cancer cells. After the tumors grew to 100 mm3, the animals were randomly assigned into different group (n=8). The results indicated that the antibodies targeting human CD40 showed a very significant antitumor effect (p<0.001). Combination of anti-CD40 and anti-PD-1 is shown more significant anti-Tumor effect. (Cooperation with CrownBio)

Fig 5. In vivo validation of anti-tumor efficacy in a MC38 tumor-bearing model of humanized CD40 mice. Homozygous humanized CD40 mice were inoculated with MC38 colon cancer cells. The results showed that an anti-human CD40 antibody exerted a very significant anti-tumor effect , demonstrating that the humanized CD40 mice are a good in vivo model for validating the efficacy of antibodies targeting human CD40. Mean volume ± SEM of tumor tissues(A). Mean body weight ± SEM of mice(B).

Fig 6. Tumor and weight changes over treated with anti-human CD40 antibody.

Fig 7. Complete blood count (CBC) of anti-human CD40-antibody treated hCD40 mice.

Fig 8.Blood chemistry of anti-human CD40-antibody treated hCD40 mice.

Fig 9. Cytokine analysis of anti-human CD40 antibody treated hCD40 mice.?

Cytokine analysis of anti-human CD40 antibody treated MC38 tumor-bearing model of humanized CD40. Anti-human CD40 antibody treatment led to significant increase several cytokines including IL-12(p40), Eotaxin, etc.