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Updated: Mar 17, 2026
Testing Cancer Immunotherapeutics in a Humanized Mouse Model Bearing Human Tumors
Published on: December 16, 2022
Antitumor Effect of Programmed Death-1 (PD-1) Blockade in Humanized the NOG-MHC Double Knockout Mouse
Tadashi Ashizawa1, Akira Iizuka1, Chizu Nonomura1
1Immunotherapy Division, Shizuoka Cancer Center Research Institute, Nagaizumi-cho, Sunto-gun, Shizuoka, Japan.
Purpose:
Humanized mouse models using NOD/Shi-scid-IL2rγnull (NOG) and NOD/LtSz-scid IL2rγnull (NSG) mouse are associated with several limitations, such as long incubation time for stem cell engraftment and the development of xenograft versus host disease in mice injected with peripheral blood mononuclear cells (PBMCs). To solve problems, we used humanized major histocompatibility class I- and class II-deficient NOG mice (referred to as NOG-dKO) to evaluate the antitumor effect of anti-programmed death-1 (PD-1) antibody.
Experimental Design:
Humanized NOG-dKO mice, in which human PBMCs and human lymphoma cell line SCC-3, or glioblastoma cell line U87 were transplanted, were used as an immunotherapy model to investigate the effect of anti-PD-1 antibody. A biosimilar anti-PD-1 mAb generated in our laboratory was administered to humanized NOG-dKO mice transplanted with tumors.
Results:
Within 4 weeks after transplantation, human CD45+ cells in antibody-treated mice constituted approximately 70% of spleen cells. The injection of anti-PD-1 antibody reduced by more 50% the size of SCC-3 and U87 tumors. In addition, induction of CTLs against SCC-3 cells and upregulation of natural killer cell activity was observed in the antibody-treated group. Tumor-infiltrating lymphocyte profiling showed that more exhausted marker (PD1+TIM3+LAG3+) positive T cells maintained in anti-PD-1 antibody-treated tumor. A greater number of CD8+ and granzyme-producing T cells infiltrated the tumor in mice treated with the anti-PD-1 antibody.
Conclusions:
These results suggest that NOG-dKO mice might serve as a good humanized immunotherapy model to evaluate the efficacy of anti-PD-1 antibody prior to the clinical treatment. Clin Cancer Res; 23(1); 149-58. ©2016 AACR.
Insights
Humanized NOG-dKO mice effectively model anti-programmed death-1 (PD-1) antibody therapy. This model demonstrated significant tumor reduction and enhanced immune cell activity, supporting its use in preclinical immunotherapy evaluation.
Area of Science:
- Immunology
- Cancer Research
- Preclinical Models
Background:
- Existing humanized mouse models (NOG, NSG) have limitations including long engraftment times and xenograft versus host disease.
- These limitations hinder the efficient evaluation of immunotherapies.
Purpose of the Study:
- To evaluate the efficacy of anti-programmed death-1 (PD-1) antibody in a novel humanized major histocompatibility class I- and class II-deficient NOG (NOG-dKO) mouse model.
- To establish a more effective preclinical model for immunotherapy research.
Main Methods:
- Humanized NOG-dKO mice were established by transplanting human peripheral blood mononuclear cells (PBMCs) and human tumor cell lines (SCC-3 lymphoma, U87 glioblastoma).
- The antitumor effect of a biosimilar anti-PD-1 monoclonal antibody (mAb) was assessed in these transplanted mice.
Main Results:
- Anti-PD-1 antibody treatment led to approximately 70% human CD45+ cells in spleen within 4 weeks.
- Tumor volumes of SCC-3 and U87 were reduced by over 50% in antibody-treated mice.
- Enhanced immune responses observed included induction of cytotoxic T lymphocytes (CTLs), increased natural killer (NK) cell activity, and greater infiltration of CD8+ and granzyme-producing T cells into tumors.
Conclusions:
- NOG-dKO mice provide a robust humanized model for evaluating anti-PD-1 antibody efficacy.
- This model shows promise for preclinical assessment of immunotherapies before clinical application.
