hPD-1

• Data from flow cytometry (FACS) analysis

  

Fig1 . Expression of PD-1 in the activated spleen lymphocytes of humanized PD-1 Homozygous mice is detected by FACS.

Fig 2.Frequency of leukocyte and myeloid subpopulations in the spleen by flow cytometry

Spleen was collected from WT C57BL/6 mice and HO hPD-1 mice (12-week-old, female, n=3). Flow cytometry analysis of frequencies of (A) lymphocyte and (B) myeloid subpopulations in the spleen. Bar indicated expressed as mean ± SEM.

Abbr.  HO, homozygous; WT, wild type.

Fig 3.Frequency of leukocyte and myeloid subpopulations in the blood by flow cytometry

Blood was collected from WT C57BL/6 mice and HO hPD-1 mice (12-week-old, female, n=3). Flow cytometry analysis of frequencies of (A) lymphocyte and (B) myeloid subpopulations in the blood. Bar indicated expressed as mean ± SEM.

Abbr.  HO, homozygous; WT, wild type.

• In vivo validation in a MC38 tumor-bearing model of humanized PD-1 mice

Fig2 . In vivo anti-tumor effect of an anti-human PD-1 antibody in a humanized mouse model of PD-1. Anti-human-PD-1 drugs significantly inhibited the growth of MC38 tumors in PD-1 mice, demonstrating that the humanized PD-1 mice can be used to assess the anti-human PD-1 antibody. 

A. Mean volume ± SEM of tumor tissues (completed in cooperation with Genscript). 

In vivo validation of anti-tumor efficacy in a MC38 tumor-bearing model of humanized PD-1 mice. Homozygous humanized PD-1 mice were inoculated with MC38 colon cancer cells. After the tumors grew to 100 mm3, the animals were randomly assigned into a control group and a treatment group (n=8). The drug was given twice a week for a total of 4 administrations. The results showed that Keytruda, a drug targeting human PD-1, exerted a very significant anti-tumor effect (p<0.001), demonstrating that the humanized PD-1 mice are a good in vivo model for validating the efficacy of antibodies targeting human PD-1. 

B. Mean volume ± SEM of tumor tissues. C. Mean body weight ± SEM of mice (data were obtained in cooperation with PharmaLegacy).

In vivo dose validation of anti-tumor efficacy in a MC38 tumor-bearing model of humanized PD-1 mice. Homozygous humanized PD-1 mice were inoculated with MC38 colon cancer cells. After the tumors grew to about 90 mm3, the animals were randomly assigned into a control group and a treatment group (n=9). The results showed that the antibodies targeting human PD-1 showed a very significant antitumor effect (p<0.001), and such antitumor effect is dose-dependent.

Fig3 . In vivo dose validation in a MC38 tumor-bearing model of humanized PD-1 mice. Homozygous humanized PD-1 mice were inoculated with MC38 colon cancer cells. After the tumors grew to 100 mm3, the animals were randomly assigned into a control group and a treatment group (n=8). The drug was given twice a week for a total of 4 administrations.

Case Study

As one of the first five anti-PD-1 drugs with BLAs accepted by CFDA, Sintilimab has shown potent T cell stimulating activity and significant anti-tumor efficacy. (read more)

Fig4 .  In vivo hPD-1 knock-in mouse model to test anti-tumor efficacy of Sintilimab. (A) Tumor growth inhibition (TGI) of MC38 tumors in hPD-1 knock-in mice of individual animals treated with different doses of Sintilimab. (B) Effect of Sintilimab on percentage changes in mouse body weight(mean). (C) Changes in ratios of tumor infiltrating CD4+, CD8+ and Treg cells. For d8: IgG (n = 2); Sintilimab (n = 2). For d14, n = 3 for all groups. P values were calculated using a two-tailed t-test method.

Fig5. Means of tumor volume(A) and body weight(B). PD-1 humanized mouse model implanted with MC38-hCLDN18.2 tumors to evaluate the in vivo antitumor efficacy of anti-hCLDN18.2 mAb alone and in combination with Keytruda. The anti-hCLDN18.2 mAb in combination with Keytruda significantly inhibited the growth of MC38-hCLDN18.2 tumors in PD-1 humanized mice, demonstrating that this model is suitable for evaluating the in vivo efficacy of anti-hCLDN18.2 mAb alone and in combination with PD-1/PD-L1 antibodies. n=8. Mean士SEM. t-test,*P < 0.05.