{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,4,30]],"date-time":"2025-04-30T04:23:21Z","timestamp":1745987001387,"version":"3.40.4"},"reference-count":45,"publisher":"Georg Thieme Verlag KG","issue":"05","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2025,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>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.<\/jats:p><jats:p>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.<\/jats:p><jats:p>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.<\/jats:p>","DOI":"10.1055\/a-2518-7157","type":"journal-article","created":{"date-parts":[[2025,4,29]],"date-time":"2025-04-29T22:30:31Z","timestamp":1745965831000},"page":"435-446","source":"Crossref","is-referenced-by-count":0,"title":["Generation of a Severe Hemophilia A Humanized Mouse Model Capable of Inducing an Anti-FVIII Immune Response"],"prefix":"10.1055","volume":"125","author":[{"given":"Akihisa","family":"Oda","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shoko","family":"Furukawa","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahiro","family":"Kitabatake","sequence":"additional","affiliation":[{"name":"Department of Immunology, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Noriko","family":"Ouji-Sageshima","sequence":"additional","affiliation":[{"name":"Department of Immunology, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshihiro","family":"Ito","sequence":"additional","affiliation":[{"name":"Department of Immunology, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Riichi","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takeshi","family":"Kawamura","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuto","family":"Nakajima","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"},{"name":"Advanced Medical Science of Thrombosis and Hemostasis, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0157-5215","authenticated-orcid":false,"given":"Naruto","family":"Shimonishi","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"},{"name":"The Course of Thrombosis and Hemostasis Molecular Pathology, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenichi","family":"Ogiwara","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Midori","family":"Shima","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"},{"name":"Thrombosis and Hemostasis Research Center, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keiji","family":"Nogami","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"194","published-online":{"date-parts":[[2025,4,29]]},"reference":[{"issue":"06","key":"ref1","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1056\/NEJMoa067659","article-title":"Prophylaxis versus episodic treatment to prevent joint disease in boys with severe hemophilia","volume":"357","author":"M J Manco-Johnson","year":"2007","journal-title":"N Engl J Med"},{"issue":"23","key":"ref2","doi-asserted-by":"crossref","first-page":"2559","DOI":"10.1182\/blood-2017-05-782912","article-title":"Marginal zone B cells are critical to factor VIII inhibitor formation in mice with hemophilia A","volume":"130","author":"P E Zerra","year":"2017","journal-title":"Blood"},{"issue":"11","key":"ref3","doi-asserted-by":"crossref","first-page":"1816","DOI":"10.1111\/j.1538-7836.2009.03571.x","article-title":"Splenic marginal zone antigen-presenting cells are critical for the primary allo-immune response to therapeutic factor VIII in hemophilia A","volume":"7","author":"A Navarrete","year":"2009","journal-title":"J Thromb Haemost"},{"issue":"22","key":"ref4","doi-asserted-by":"crossref","first-page":"e159925","DOI":"10.1172\/JCI159925","article-title":"Immune tolerance against infused FVIII in hemophilia A is mediated by PD-L1+ Tregs","volume":"132","author":"J Becker-Gotot","year":"2022","journal-title":"J Clin Invest"},{"issue":"20","key":"ref5","doi-asserted-by":"crossref","first-page":"3099","DOI":"10.1182\/bloodadvances.2019000650","article-title":"Induction of activated T follicular helper cells is critical for anti-FVIII inhibitor development in hemophilia A mice","volume":"3","author":"W Jing","year":"2019","journal-title":"Blood Adv"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.thromres.2023.03.003","article-title":"The spleen is the major site for the development and expansion of inhibitor producing-cells in hemophilia A mice upon FVIII infusion developing high-titer inhibitor","volume":"231","author":"A Oda","year":"2023","journal-title":"Thromb Res"},{"issue":"06","key":"ref7","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1055\/s-0037-1613929","article-title":"Humanized severe combined immunodeficient mice as a potential model for the study of tolerance to factor VIII","volume":"83","author":"B Vanzieleghem","year":"2000","journal-title":"Thromb Haemost"},{"key":"ref8","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.thromres.2024.01.019","article-title":"Recapitulating the immune system of hemophilia A patients with inhibitors using immunodeficient mice","volume":"235","author":"S C Chou","year":"2024","journal-title":"Thromb Res"},{"issue":"09","key":"ref9","doi-asserted-by":"crossref","first-page":"3175","DOI":"10.1182\/blood-2001-12-0207","article-title":"NOD\/SCID\/gamma(c)(null) mouse: an excellent recipient mouse model for engraftment of human cells","volume":"100","author":"M Ito","year":"2002","journal-title":"Blood"},{"issue":"05","key":"ref10","doi-asserted-by":"crossref","first-page":"1565","DOI":"10.1182\/blood-2005-02-0516","article-title":"Development of functional human blood and immune systems in NOD\/SCID\/IL2 receptor gamma chain(null) mice","volume":"106","author":"F Ishikawa","year":"2005","journal-title":"Blood"},{"issue":"12","key":"ref11","doi-asserted-by":"crossref","first-page":"6145","DOI":"10.4049\/jimmunol.1103660","article-title":"Development of mature and functional human myeloid subsets in hematopoietic stem cell-engrafted NOD\/SCID\/IL2r\u03b3KO mice","volume":"188","author":"S Tanaka","year":"2012","journal-title":"J Immunol"},{"key":"ref12","first-page":"15.21.1","article-title":"Creation of \u201chumanized\u201d mice to study human immunity","volume":"Chapter 15","author":"T Pearson","year":"2008","journal-title":"Curr Protoc Immunol"},{"issue":"43","key":"ref13","doi-asserted-by":"crossref","first-page":"15951","DOI":"10.1073\/pnas.0604493103","article-title":"Disseminated and sustained HIV infection in CD34+ cord blood cell-transplanted Rag2-\/-gamma c-\/- mice","volume":"103","author":"S Baenziger","year":"2006","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"07","key":"ref14","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1093\/intimm\/dxp050","article-title":"The analysis of the functions of human B and T cells in humanized NOD\/shi-scid\/gammac(null) (NOG) mice (hu-HSC NOG mice)","volume":"21","author":"Y Watanabe","year":"2009","journal-title":"Int Immunol"},{"issue":"09","key":"ref15","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.1016\/j.exphem.2006.05.006","article-title":"Development of human-human hybridoma from anti-Her-2 peptide-producing B cells in immunized NOG mouse","volume":"34","author":"Y Kametani","year":"2006","journal-title":"Exp Hematol"},{"issue":"03","key":"ref16","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1182\/bloodadvances.2018023887","article-title":"Human hematopoietic stem cell maintenance and myeloid cell development in next-generation humanized mouse models","volume":"3","author":"T R Sippel","year":"2019","journal-title":"Blood Adv"},{"issue":"08","key":"ref17","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1182\/blood-2016-04-709584","article-title":"A novel humanized mouse model with significant improvement of class-switched, antigen-specific antibody production","volume":"129","author":"H Yu","year":"2017","journal-title":"Blood"},{"issue":"04","key":"ref18","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1093\/intimm\/dxs045","article-title":"Induction of human humoral immune responses in a novel HLA-DR-expressing transgenic NOD\/Shi-scid\/\u03b3cnull mouse","volume":"24","author":"M Suzuki","year":"2012","journal-title":"Int Immunol"},{"issue":"02","key":"ref19","doi-asserted-by":"crossref","first-page":"e201800195","DOI":"10.26508\/lsa.201800195","article-title":"Human NK cell development in hIL-7 and hIL-15 knockin NOD\/SCID\/IL2rgKO mice","volume":"2","author":"M Matsuda","year":"2019","journal-title":"Life Sci Alliance"},{"issue":"7950","key":"ref20","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1038\/s41586-023-05739-9","article-title":"Chemically defined cytokine-free expansion of human haematopoietic stem cells","volume":"615","author":"M Sakurai","year":"2023","journal-title":"Nature"},{"issue":"01","key":"ref21","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1186\/s13287-024-03799-w","article-title":"Improvement of multilineage hematopoiesis in hematopoietic stem cell-transferred c-kit mutant NOG-EXL humanized mice","volume":"15","author":"R Ito","year":"2024","journal-title":"Stem Cell Res Ther"},{"issue":"08","key":"ref22","doi-asserted-by":"crossref","first-page":"1489","DOI":"10.1038\/s41590-024-01880-3","article-title":"A humanized mouse that mounts mature class-switched, hypermutated and neutralizing antibody responses","volume":"25","author":"D P Chupp","year":"2024","journal-title":"Nat Immunol"},{"issue":"23","key":"ref23","doi-asserted-by":"crossref","first-page":"5532","DOI":"10.1182\/blood-2011-07-367680","article-title":"Role of bone marrow transplantation for correcting hemophilia A in mice","volume":"119","author":"A Follenzi","year":"2012","journal-title":"Blood"},{"issue":"07","key":"ref24","doi-asserted-by":"crossref","first-page":"881","DOI":"10.3324\/haematol.2014.123117","article-title":"Extrahepatic sources of factor VIII potentially contribute to the coagulation cascade correcting the bleeding phenotype of mice with hemophilia A","volume":"100","author":"D Zanolini","year":"2015","journal-title":"Haematologica"},{"issue":"08","key":"ref25","doi-asserted-by":"crossref","first-page":"2452","DOI":"10.1038\/s41596-019-0187-x","article-title":"Creation of CRISPR-based germline-genome-engineered mice without ex vivo handling of zygotes by i-GONAD","volume":"14","author":"C B Gurumurthy","year":"2019","journal-title":"Nat Protoc"},{"issue":"03","key":"ref26","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1055\/s-0037-1613237","article-title":"Expression and characterization of recombinant murine factor VIII","volume":"88","author":"C Doering","year":"2002","journal-title":"Thromb Haemost"},{"issue":"01","key":"ref27","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1038\/ng0595-119","article-title":"Targeted disruption of the mouse factor VIII gene produces a model of haemophilia A","volume":"10","author":"L Bi","year":"1995","journal-title":"Nat Genet"},{"issue":"02","key":"ref28","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1111\/jth.13202","article-title":"Characterization of a genetically engineered mouse model of hemophilia A with complete deletion of the F8 gene","volume":"14","author":"B N Chao","year":"2016","journal-title":"J Thromb Haemost"},{"issue":"04","key":"ref29","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1002\/iid3.124","article-title":"Improved B cell development in humanized NOD-scid IL2R\u03b3null\n           mice transgenically expressing human stem cell factor, granulocyte-macrophage colony-stimulating factor and interleukin-3","volume":"4","author":"S Jangalwe","year":"2016","journal-title":"Immun Inflamm Dis"},{"issue":"03","key":"ref30","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1538\/expanim.63.321","article-title":"NOD-Rag2null IL-2R\u03b3null mice: an alternative to NOG mice for generation of humanized mice","volume":"63","author":"I Katano","year":"2014","journal-title":"Exp Anim"},{"issue":"24","key":"ref31","doi-asserted-by":"crossref","first-page":"3706","DOI":"10.1182\/blood-2014-02-555151","article-title":"A conditional knockout mouse model reveals endothelial cells as the principal and possibly exclusive source of plasma factor VIII","volume":"123","author":"S A Fahs","year":"2014","journal-title":"Blood"},{"issue":"01","key":"ref32","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1182\/blood-2003-05-1684","article-title":"Transplanted adult hematopoietic stems cells differentiate into functional endothelial cells","volume":"103","author":"A S Bailey","year":"2004","journal-title":"Blood"},{"issue":"11","key":"ref33","doi-asserted-by":"crossref","first-page":"1527","DOI":"10.1172\/JCI8296","article-title":"Transplanted cord blood-derived endothelial precursor cells augment postnatal neovascularization","volume":"105","author":"T Murohara","year":"2000","journal-title":"J Clin Invest"},{"issue":"05","key":"ref34","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1055\/s-0038-1656061","article-title":"High prevalence of elevated factor VIII levels in patients referred for thrombophilia screening: role of increased synthesis and relationship to the acute phase reaction","volume":"77","author":"J O'Donnell","year":"1997","journal-title":"Thromb Haemost"},{"issue":"06","key":"ref35","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1111\/hae.13852","article-title":"Individual combinations of danger signals synergistically increase FVIII product immunogenicity","volume":"25","author":"L Miller","year":"2019","journal-title":"Haemophilia"},{"issue":"02","key":"ref36","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1111\/j.1365-2516.2009.02122.x","article-title":"New early prophylaxis regimen that avoids immunological danger signals can reduce FVIII inhibitor development","volume":"16","author":"K Kurnik","year":"2010","journal-title":"Haemophilia"},{"issue":"04","key":"ref37","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1055\/s-0038-1637734","article-title":"Human dendritic cells synergistically activated by FVIII plus LPS induce activation of autologous CD4+ T cells","volume":"118","author":"L Miller","year":"2018","journal-title":"Thromb Haemost"},{"issue":"04","key":"ref38","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1016\/j.cell.2006.02.015","article-title":"Pathogen recognition and innate immunity","volume":"124","author":"S Akira","year":"2006","journal-title":"Cell"},{"issue":"23","key":"ref39","doi-asserted-by":"crossref","first-page":"4886","DOI":"10.1182\/blood-2009-10-246678","article-title":"Age-dependent vulnerability to endotoxemia is associated with reduction of anticoagulant factors activated protein C and thrombomodulin","volume":"115","author":"M E Starr","year":"2010","journal-title":"Blood"},{"issue":"07","key":"ref40","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1046\/j.1538-7836.2003.00261.x","article-title":"Inflammation and thrombosis","volume":"1","author":"C T Esmon","year":"2003","journal-title":"J Thromb Haemost"},{"issue":"08","key":"ref41","doi-asserted-by":"crossref","first-page":"e1011603","DOI":"10.1371\/journal.ppat.1011603","article-title":"Germline-encoded specificities and the predictability of the B cell response","volume":"19","author":"M C Vieira","year":"2023","journal-title":"PLoS Pathog"},{"key":"ref42","doi-asserted-by":"crossref","first-page":"13376","DOI":"10.1038\/ncomms13376","article-title":"Germline-encoded neutralization of a Staphylococcus aureus virulence factor by the human antibody repertoire","volume":"7","author":"Y A Yeung","year":"2016","journal-title":"Nat Commun"},{"issue":"6507","key":"ref43","doi-asserted-by":"crossref","first-page":"1119","DOI":"10.1126\/science.abd2321","article-title":"Structural basis of a shared antibody response to SARS-CoV-2","volume":"369","author":"M Yuan","year":"2020","journal-title":"Science"},{"issue":"12","key":"ref44","doi-asserted-by":"crossref","first-page":"2922","DOI":"10.1182\/blood-2011-09-379453","article-title":"F8 gene mutation type and inhibitor development in patients with severe hemophilia A: systematic review and meta-analysis","volume":"119","author":"S C Gouw","year":"2012","journal-title":"Blood"},{"issue":"01","key":"ref45","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1186\/s13059-018-1400-x","article-title":"i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases","volume":"19","author":"M Ohtsuka","year":"2018","journal-title":"Genome Biol"}],"container-title":["Thrombosis and Haemostasis"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.thieme-connect.de\/products\/ejournals\/pdf\/10.1055\/a-2518-7157.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,29]],"date-time":"2025-04-29T22:31:10Z","timestamp":1745965870000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.1055\/a-2518-7157"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,29]]},"references-count":45,"journal-issue":{"issue":"05","published-online":{"date-parts":[[2025,4,29]]},"published-print":{"date-parts":[[2025,5]]}},"URL":"https:\/\/doi.org\/10.1055\/a-2518-7157","archive":["Portico","CLOCKSS"],"relation":{},"ISSN":["0340-6245","2567-689X"],"issn-type":[{"value":"0340-6245","type":"print"},{"value":"2567-689X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,29]]}}}