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

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.

Publikationsverlauf

Eingereicht: 05. November 2024

Angenommen: 16. Januar 2025

Artikel online veröffentlicht:
29. April 2025

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