title: Transient low-dose methotrexate generates B regulatory cells that mediate antigen-specific tolerance to alglucosidase alfa.
abstract: Biologic drugs, including enzyme-replacement therapies, can elicit anti-drug Abs (ADA) that may interfere with drug efficacy and impact patient safety. In an effort to control ADA, we focused on identifying regimens of immune tolerance induction that may be readily available for clinical use. Data generated in both wild-type mice and a Pompe disease mouse model demonstrate that single-cycle, low-dose methotrexate can be as effective as three cycles of methotrexate in providing a long-lived reduction in alglucosidase alfa-specific ADA. In addition, we show that methotrexate induces Ag-specific tolerance as mice generate similar Ab responses to an irrelevant Ag regardless of prior methotrexate treatment. Methotrexate-induced immune tolerance does not seem to involve cell depletion, but rather a specific expansion of IL-10- and TGF-β-secreting B cells that express Foxp3, suggesting an induction of regulatory B cells. The mechanism of immune tolerance induction appears to be IL-10 dependent, as methotrexate does not induce immune tolerance in IL-10 knockout mice. Splenic B cells from animals that have been tolerized to alglucosidase alfa with methotrexate can transfer tolerance to naive hosts. We hypothesize that methotrexate induction treatment concomitant with initial exposure to the biotherapeutic can induce Ag-specific immune tolerance in mice through a mechanism that appears to involve the induction of regulatory B cells.
authors.affiliation: Genzyme, a Sanofi company, Framingham, MA 01701.
authors.forename: Marguerite S
authors.initials: MS
authors.lastname: Joly
authors.affiliation: Genzyme, a Sanofi company, Framingham, MA 01701.
authors.forename: Roderick P
authors.initials: RP
authors.lastname: Martin
authors.affiliation: Genzyme, a Sanofi company, Framingham, MA 01701.
authors.forename: Shibani
authors.initials: S
authors.lastname: Mitra-Kaushik
authors.affiliation: Genzyme, a Sanofi company, Framingham, MA 01701.
authors.forename: Lucy
authors.initials: L
authors.lastname: Phillips
authors.affiliation: Genzyme, a Sanofi company, Framingham, MA 01701.
authors.forename: Alida
authors.initials: A
authors.lastname: D'Angona
authors.affiliation: Genzyme, a Sanofi company, Framingham, MA 01701.
authors.forename: Susan M
authors.initials: SM
authors.lastname: Richards
authors.affiliation: Genzyme, a Sanofi company, Framingham, MA 01701 alexandra.joseph@genzyme.com.
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: Joseph
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D055610:Antigens, CD1d / Q000276:immunology; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D060151:B-Lymphocytes, Regulatory / Q000187:drug effects / Q000276:immunology* / Q000637:transplantation; D018956:CD5 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D051858:Forkhead Transcription Factors / Q000096:biosynthesis; D007108:Immune Tolerance / Q000187:drug effects*; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D008727:Methotrexate / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D050378:T-Lymphocytes, Regulatory / Q000276:immunology; D016212:Transforming Growth Factor beta / Q000378:metabolism; D000520:alpha-Glucosidases / Q000276:immunology*
chemical_list: D055610:Antigens, CD1d; D000964:Antimetabolites, Antineoplastic; C584692:CD1d antigen, mouse; D018956:CD5 Antigens; C526836:Cd5 protein, mouse; D005493:Folic Acid Antagonists; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; C556062:IL10 protein, mouse; D007074:Immunoglobulin G; D016212:Transforming Growth Factor beta; D016753:Interleukin-10; C509951:GAA protein, human; D000520:alpha-Glucosidases; D008727:Methotrexate