title: Plant-based oral tolerance to hemophilia therapy employs a complex immune regulatory response including LAP+CD4+ T cells.
abstract: Coagulation factor replacement therapy for the X-linked bleeding disorder hemophilia is severely complicated by antibody ("inhibitor") formation. We previously found that oral delivery to hemophilic mice of cholera toxin B subunit-coagulation factor fusion proteins expressed in chloroplasts of transgenic plants suppressed inhibitor formation directed against factors VIII and IX and anaphylaxis against factor IX (FIX). This observation and the relatively high concentration of antigen in the chloroplasts prompted us to evaluate the underlying tolerance mechanisms. The combination of oral delivery of bioencapsulated FIX and intravenous replacement therapy induced a complex, interleukin-10 (IL-10)-dependent, antigen-specific systemic immune suppression of pathogenic antibody formation (immunoglobulin [Ig] 1/inhibitors, IgE) in hemophilia B mice. Tolerance induction was also successful in preimmune mice but required prolonged oral delivery once replacement therapy was resumed. Orally delivered antigen, initially targeted to epithelial cells, was taken up by dendritic cells throughout the small intestine and additionally by F4/80(+) cells in the duodenum. Consistent with the immunomodulatory responses, frequencies of tolerogenic CD103(+) and plasmacytoid dendritic cells were increased. Ultimately, latency-associated peptide expressing CD4(+) regulatory T cells (CD4(+)CD25(-)LAP(+) cells with upregulated IL-10 and transforming growth factor-β (TGF-β) expression) as well as conventional CD4(+)CD25(+) regulatory T cells systemically suppressed anti-FIX responses.
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL;
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Wang
authors.affiliation: Departments of Biochemistry and Pathology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA;
authors.forename: Jin
authors.initials: J
authors.lastname: Su
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL;
authors.forename: Alexandra
authors.initials: A
authors.lastname: Sherman
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL;
authors.forename: Geoffrey L
authors.initials: GL
authors.lastname: Rogers
authors.affiliation: Division of Immunology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston MA; and.
authors.forename: Gongxian
authors.initials: G
authors.lastname: Liao
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL;
authors.forename: Brad E
authors.initials: BE
authors.lastname: Hoffman
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY.
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
authors.affiliation: Division of Immunology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston MA; and.
authors.forename: Cox
authors.initials: C
authors.lastname: Terhorst
authors.affiliation: Departments of Biochemistry and Pathology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA;
authors.forename: Henry
authors.initials: H
authors.lastname: Daniell
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL;
authors.forename: Roland W
authors.initials: RW
authors.lastname: Herzog
mesh_terms: D000284:Administration, Oral; D019264:Adoptive Transfer; D000818:Animals; D000917:Antibody Formation; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D005164:Factor IX / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D002836:Hemophilia B / Q000276:immunology / Q000628:therapy*; D006801:Humans; D016753:Interleukin-10 / Q000276:immunology; D008297:Male; D051379:Mice; D008517:Phytotherapy; D030821:Plants, Genetically Modified / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D014026:Nicotiana / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000276:immunology
chemical_list: D011993:Recombinant Fusion Proteins; D016212:Transforming Growth Factor beta; D016753:Interleukin-10; D005164:Factor IX