Enhanced diversity of human immune cells in a PBMC-engrafted NSG-SGM3-IL15-DKO (S15-DKO) immunodeficient mouse model
- 1Department of Innovation & Product Development, The Jackson Laboratory
- 2Diabetes Center of Excellence, University of Massachusetts Chan Medical School
Abstract
Background: Peripheral blood mononuclear cell (PBMC)-humanized mouse models are critical tools for preclinical evaluation of cancer immunotherapies, yet current models offer limited recapitulation of human immune diversity, particularly myeloid and natural killer (NK) cells. To address this limitation, we generated a novel immunocompromised mouse strain NSG-SGM3-IL15-DKO (S15-DKO) combining the NSG-MHC I/II double-knockout (DKO), which delays xenogeneic graft-versus-host disease, with the NSG-SGM3-IL15 human cytokine-expressing strain. Here, we evaluated PBMC engraftment and established a humanized immune-oncology model to test bispecific T cell engager (BiTE) efficacy and cytokine release in S15-DKO mice compared to DKO mice.
Methods: S15-DKO and DKO mice were engrafted with 1x107 human PBMCs from various healthy donors, with or without prior exposure to Raji lymphoma cells or irradiation. Retro-orbital (RO) blood (and spleen) was collected to assess human immune cells engraftment by flow cytometry. Irradiated, Raji-bearing S15-DKO and DKO mice were treated with PBS or anti-CD3xCD19 BiTE. Bioluminescence imaging was performed to measure tumor burden, and plasma was collected to measure cytokine levels.
Results: S15-DKO mice engrafted with PBMCs from multiple donors supported significantly higher levels of human CD45 ⁺ cells, CD4 ⁺ and CD8 ⁺ T cells, and NK cells compared to DKO mice without the need for irradiation, with body weight changes and survival comparable to DKO controls. B cells, dendritic cells, and macrophages also showed significantly improved engraftment in S15-DKO mice compared to DKO mice with certain PBMC donors. Engraftment of PBMCs in S15-DKO mice bearing Raji lymphoma cells was also higher compared to DKO mice, with significantly elevated numbers of NK cells and dendritic cells in addition to T cells. Preconditioning the S15-DKO mice with irradiation prior to PBMC engraftment further enhanced immune cell reconstitution, accelerating engraftment kinetics and increasing absolute cell counts across multiple immune subsets. In a therapeutic model, irradiated S15-DKO mice bearing Raji-luc cells demonstrated superior tumor reduction and significantly higher levels of cytokine release compared to DKO mice following anti-CD3xCD19 BiTE treatment.
Discussion: These findings establish PBMC-humanized S15-DKO mice as a novel preclinical platform for evaluating efficacy and cytokine release of immunotherapies that require diverse populations of human immune cells.
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BibTeX
@article{zheng2026enhanced,
title = {{Enhanced diversity of human immune cells in a PBMC-engrafted NSG-SGM3-IL15-DKO (S15-DKO) immunodeficient mouse model}},
author = {Xiaoqing Zheng and Michael A. Brehm and Leandro Salati D’Abronzo and Guoxiang Yang and Jessica K. Wong and Beau Parry and Jiwon Yang and James G. Keck and Li-Chin Yao},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1967450},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1967450/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Enhanced diversity of human immune cells in a PBMC-engrafted NSG-SGM3-IL15-DKO (S15-DKO) immunodeficient mouse model
AU - Xiaoqing Zheng
AU - Michael A. Brehm
AU - Leandro Salati D’Abronzo
AU - Guoxiang Yang
AU - Jessica K. Wong
AU - Beau Parry
AU - Jiwon Yang
AU - James G. Keck
AU - Li-Chin Yao
PY - 2026
DA - 2026/10/27
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1967450
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1967450/full
AB - Background: Peripheral blood mononuclear cell (PBMC)-humanized mouse models are critical tools for preclinical evaluation of cancer immunotherapies, yet current models offer limited recapitulation of human immune diversity, particularly myeloid and natural killer (NK) cells. To address this limitation, we generated a novel immunocompromised mouse strain NSG-SGM3-IL15-DKO (S15-DKO) combining the NSG-MHC I/II double-knockout (DKO), which delays xenogeneic graft-versus-host disease, with the NSG-SGM3-IL15 human cytokine-expressing strain. Here, we evaluated PBMC engraftment and established a humanized immune-oncology model to test bispecific T cell engager (BiTE) efficacy and cytokine release in S15-DKO mice compared to DKO mice. Methods: S15-DKO and DKO mice were engrafted with 1x107 human PBMCs from various healthy donors, with or without prior exposure to Raji lymphoma cells or irradiation. Retro-orbital (RO) blood (and spleen) was collected to assess human immune cells engraftment by flow cytometry. Irradiated, Raji-bearing S15-DKO and DKO mice were treated with PBS or anti-CD3xCD19 BiTE. Bioluminescence imaging was performed to measure tumor burden, and plasma was collected to measure cytokine levels. Results: S15-DKO mice engrafted with PBMCs from multiple donors supported significantly higher levels of human CD45 ⁺ cells, CD4 ⁺ and CD8 ⁺ T cells, and NK cells compared to DKO mice without the need for irradiation, with body weight changes and survival comparable to DKO controls. B cells, dendritic cells, and macrophages also showed significantly improved engraftment in S15-DKO mice compared to DKO mice with certain PBMC donors. Engraftment of PBMCs in S15-DKO mice bearing Raji lymphoma cells was also higher compared to DKO mice, with significantly elevated numbers of NK cells and dendritic cells in addition to T cells. Preconditioning the S15-DKO mice with irradiation prior to PBMC engraftment further enhanced immune cell reconstitution, accelerating engraftment kinetics and increasing absolute cell counts across multiple immune subsets. In a therapeutic model, irradiated S15-DKO mice bearing Raji-luc cells demonstrated superior tumor reduction and significantly higher levels of cytokine release compared to DKO mice following anti-CD3xCD19 BiTE treatment. Discussion: These findings establish PBMC-humanized S15-DKO mice as a novel preclinical platform for evaluating efficacy and cytokine release of immunotherapies that require diverse populations of human immune cells.
SN - 1664-3224
ER -