Synergistic pathogenic effects of combined mouse monoclonal anti-desmoglein 3 IgG antibodies on pemphigus vulgaris blister formation.
Pemphigus vulgaris (PV) is an autoimmune blistering disease caused by anti-desmoglein 3 (Dsg3) IgG antibodies. Previously, we generated an active mouse model for PV by adoptive transfer of splenocytes from immunized or naive Dsg3(-/-) mice. In this study, we isolated 10 anti-Dsg3 IgG mAbs (NAK-series) from PV model mice generated by transfer of naive Dsg3(-/-) splenocytes. We characterized their epitopes using domain-swapped and point-mutated Dsg1/Dsg3 molecules and examined their pathogenic activities in blister formation in three different assays. In a passive transfer model using neonatal mice, eight of 10 NAK mAbs showed pathogenic activity when injected together with half the minimum pathogenic dose of anti-Dsg1 IgG autoantibodies from pemphigus foliaceus (PF) patients. None of the mAbs could induce the PV phenotype when individual hybridoma clones were inoculated by peritoneal injection into adult Rag2(-/-) mice. NAK mAbs displayed a range of potency in an in vitro dissociation assay using primary cultured mouse keratinocytes. Interestingly, when multiple hybridoma clones recognizing different epitopes were inoculated in combination, recipient mice developed the PV phenotype. In vitro dissociation assays confirmed that combined NAK mAbs had synergistic pathogenic effects. These findings indicate that although an individual anti-Dsg3 IgG is not sufficient to cause blistering in adult mice, several together can induce the PV phenotype. These mAbs will provide a valuable tool to investigate the molecular mechanisms of blister formation, mimicking the effects of the polyclonal IgG antibodies found in patients.
Kawasaki
Hiroshi
H
Department of Dermatology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
Tsunoda
Kazuyuki
K
Hata
Tsuyoshi
T
Ishii
Ken
K
Yamada
Taketo
T
Amagai
Masayuki
M
D000375:Aging; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001201:Ascites / Q000276:immunology; D001768:Blister / Q000276:immunology*; D051184:Desmoglein 3 / Q000276:immunology*; D004338:Drug Combinations; D004357:Drug Synergism; D000939:Epitopes; D007116:Immunization, Passive; D007074:Immunoglobulin G / Q000276:immunology*; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D051379:Mice; D018345:Mice, Knockout; D010392:Pemphigus / Q000276:immunology*; D010641:Phenotype
D000911:Antibodies, Monoclonal; D051184:Desmoglein 3; D004338:Drug Combinations; D000939:Epitopes; D007074:Immunoglobulin G