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Thromb Haemost 2025; 125(05): 435-446
DOI: 10.1055/a-2518-7157
Coagulation and Fibrinolysis

Generation of a Severe Hemophilia A Humanized Mouse Model Capable of Inducing an Anti-FVIII Immune Response

Akihisa Oda
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Shoko Furukawa
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Masahiro Kitabatake
2   Department of Immunology, Nara Medical University, Kashihara, Nara, Japan
,
Noriko Ouji-Sageshima
2   Department of Immunology, Nara Medical University, Kashihara, Nara, Japan
,
Toshihiro Ito
2   Department of Immunology, Nara Medical University, Kashihara, Nara, Japan
,
Riichi Takahashi
3   Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan
,
Takeshi Kawamura
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Yuto Nakajima
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
4   Advanced Medical Science of Thrombosis and Hemostasis, Nara Medical University, Kashihara, Nara, Japan
,
Naruto Shimonishi 
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
5   The Course of Thrombosis and Hemostasis Molecular Pathology, Nara Medical University, Kashihara, Nara, Japan
,
Kenichi Ogiwara
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Midori Shima
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
6   Thrombosis and Hemostasis Research Center, Nara Medical University, Kashihara, Nara, Japan
,
Keiji Nogami
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
› Author Affiliations
Funding This work was supported by a Grant-in-Aid for Scientific Research (KAKENHI) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) [grant numbers: 18K07182, 20H00531, and 21K07825] and funded by Sanofi S.A.
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Abstract

Background

Factor VIII (FVIII) replacement therapy induces anti-FVIII neutralizing antibodies in approximately 30% of patients with severe hemophilia A (HA). Owing to the lack of experimental systems that allow for the study of human anti-FVIII immune responses, the mechanisms underlying replacement therapy-induced anti-FVIII antibodies in HA patients remain largely unknown. Therefore, experimental systems that enable the study of human anti-FVIII immune responses are needed.

Methods

We generated severe immunodeficient NOD-scid IL-2Rnull; FVIIInull mice (NOG HA) that can serve as hosts for human cord blood (hCB) transplantation and established a HA mouse with a humanized immune system to induce the anti-FVIII responses in human immune cells in vivo.

Results and Conclusion

The proportions of immune cell subsets (CD8+ T cells, CD4+ T cells, CD19+ B cells, CD33+ macrophages, and CD56+ natural killer (NK) cells) in the bone marrow, spleen, and peripheral blood were similar between NOG HA and NOG mice 4 months after hCB transplantation. The hCB-engrafted NOG HA mice retained HA severity. To activate the anti-FVIII immune response in hCB-engrafted NOG HA mice, we administered recombinant (r)FVIII plus lipopolysaccharide (LPS) once a week for 3 months. We detected both anti-FVIII IgM and IgG in the plasma of hCB-engrafted NOG HA mice after treatment with 12 doses of rFVIII and LPS. Taken together, our humanized mice with HA maintained a severe phenotype and generated human anti-FVIII IgG antibodies in vivo, thus representing a valuable model for studying human anti-FVIII immune responses.

Keywords

humanized mice - severe hemophilia A - anti-FVIII immune response - NOG mice

Authors' Contribution

A.O. designed the study, performed the experiments, analyzed the data, interpreted the results, prepared the figures, and wrote the paper. S.F., M.K., N.O., T.I., T.K., Y.N., N.S., and K.O. performed the experiments and interpreted the data. M.K. supervised the study. R.T. provided NOG mice with a continuous supply and shared breeding rights. M.S. and K.N. acquired the research funds (Bioverativ/Sanofi) and designed the study. K.N. designed the study, supervised the study, wrote and edited the manuscript, and approved the final version for publication.


Supplementary Material



Publication History

Received: 05 November 2024

Accepted: 16 January 2025

Article published online:
29 April 2025

© 2025. Thieme. All rights reserved.

Georg Thieme Verlag KG
Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany

Comment to this article:

Inhibitor Formation in the Era of Novel Haemophilia Treatment—A Humanized Mouse Model to Answer Our Questions?Thromb Haemost 2025; 125(05): 447-448
DOI: 10.1055/a-2551-8561

 

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