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

Akihisa Oda; Shoko Furukawa; Masahiro Kitabatake; Noriko Ouji-Sageshima; Toshihiro Ito; Riichi Takahashi; Takeshi Kawamura; Yuto Nakajima; Naruto Shimonishi; Kenichi Ogiwara; Midori Shima; Keiji Nogami

doi:10.1055/a-2518-7157

Thrombosis and Haemostasis, Table of Contents

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

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

,

Shoko Furukawa

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

,

Masahiro Kitabatake

²Department of Immunology, Nara Medical University, Kashihara, Nara, Japan

,

Noriko Ouji-Sageshima

²Department of Immunology, Nara Medical University, Kashihara, Nara, Japan

,

Toshihiro Ito

²Department of Immunology, Nara Medical University, Kashihara, Nara, Japan

,

Riichi Takahashi

³Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan

,

Takeshi Kawamura

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

,

Yuto Nakajima

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

⁴Advanced Medical Science of Thrombosis and Hemostasis, Nara Medical University, Kashihara, Nara, Japan

,

Naruto Shimonishi

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

⁵The Course of Thrombosis and Hemostasis Molecular Pathology, Nara Medical University, Kashihara, Nara, Japan

,

Kenichi Ogiwara

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

,

Midori Shima

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

⁶Thrombosis and Hemostasis Research Center, Nara Medical University, Kashihara, Nara, Japan

,

Keiji Nogami

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

› Author Affiliations

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

Full Text

References

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Supplementary Material