Human IgG1 Monoclonal Antibody against Human Collagen 17 Noncollagenous 16A Domain Induces Blisters via Complement Activation in Experimental Bullous Pemphigoid Model

Li, Qiang; Ujiie, Hideyuki; Shibaki, Akihiko; Wang, Gang; Moriuchi, Reine; Qiao, Hong Jiang; Morioka, Hiroshi; Shinkuma, Satoru; Natsuga, Ken; Long, Heather A.; Nishie, Wataru; Shimizu, Hiroshi

doi:10.4049/jimmunol.1000667

Cited by 55 publications

(55 citation statements)

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“…Our previous study showed that the administration of Fab fragments to hNC16A into neonatal COL17-humanized mice inhibits the complement pathway and ameliorates skin detachment (10). We also reported that hIgG1 against hNC16A with mutations at the C1q-binding Fc portion has lower pathogenicity in neonatal COL17-humanized mice than has nonmutated hIgG1 (11). More recently, we demonstrated that passive transfer of F(ab9) 2 fragments of BP-IgG or rabbit IgG against hNC16A induces skin detachment in neonatal COL17-humanized mice (13).…”

Section: Discussionmentioning

confidence: 99%

“…The HEK293 human COL17 cells used in this study were previously established (11). Briefly, the constructed plasmid pcDNA5/FRT (Invitrogen) that had been inserted with human COL17A1 cDNA (a gift from Dr. K.B.…”

Section: Establishment Of Hek293 Human Col17 Cells and Human Col17dncmentioning

confidence: 99%

“…The human C1q-binding activity of rhIgGs and Fab-B4 was determined by C1q-binding assay by ELISA as previously described, with minor modifications (11). The rhIgGs and Fab-B4 with 2-fold serial dilution (starting concentration, 20 mg/ml) in 0.1 M sodium carbonate buffer (pH 9.5) were coated in a Microtest 96-well ELISA plate (BD Biosciences) and left overnight at 4˚C.…”

Section: C1q-binding Assaymentioning

confidence: 99%

“…The pathogenicity of IgG Ab from BP patients (BP-IgG) had been unclear because BP-IgG fails to recognize murine COL17 and to reproduce the skin detachment in wild-type mice owing to differences in the amino acid sequence between humans and mice. Our recent studies using COL17-humanized (COL17 m2/2,h+ ) mice, which lack murine COL17 but express human COL17, have directly demonstrated the pathogenicity of Abs against the NC16A of human COL17 (hNC16A), including BP-IgG, in vivo (7)(8)(9)(10)(11)(12)(13).…”

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confidence: 99%

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Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Ujiie

Sasaoka

Izumi

et al. 2014

The Journal of Immunology

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109

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Complement activation and subsequent recruitment of inflammatory cells at the dermal/epidermal junction are thought to be essential for blister formation in bullous pemphigoid (BP), an autoimmune blistering disease induced by autoantibodies against type XVII collagen (COL17); however, this theory does not fully explain the pathological features of BP. Recently, the involvement of complement-independent pathways has been proposed. To directly address the question of the necessity of the complement activation in blister formation, we generated C3-deficient COL17-humanized mice. First, we show that passive transfer of autoantibodies from BP patients induced blister formation in neonatal C3-deficient COL17-humanized mice without complement activation. By using newly generated human and murine mAbs against the pathogenic noncollagenous 16A domain of COL17 with high (human IgG1, murine IgG2), low (murine IgG1), or no (human IgG4) complement activation abilities, we demonstrate that the deposition of Abs, and not complements, is relevant to the induction of blister formation in neonatal and adult mice. Notably, passive transfer of BP autoantibodies reduced the amount of COL17 in lesional mice skin, as observed in cultured normal human keratinocytes treated with the same Abs. Moreover, the COL17 depletion was associated with a ubiquitin/proteasome pathway. In conclusion, the COL17 depletion induced by BP autoantibodies, and not complement activation, is essential for the blister formation under our experimental system.

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Section: Discussionmentioning

confidence: 99%

Section: Establishment Of Hek293 Human Col17 Cells and Human Col17dncmentioning

confidence: 99%

Section: C1q-binding Assaymentioning

confidence: 99%

mentioning

confidence: 99%

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Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Ujiie

Sasaoka

Izumi

et al. 2014

The Journal of Immunology

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109

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“…Most recently, treatment with Fab mAb fragments in a passive-transfer model using COL17-humanized mice has proved that inhibition of the complement pathway can ameliorate skin detachment (41). Moreover, IgG1 mAb with mutations at C1q binding sites, reacting against NC16A, had less potential for blister formation (42). Previous experiments suggested that complement activation has primary importance in the pathogenesis of BP.…”

Section: Discussionmentioning

confidence: 99%

Antibodies to Pathogenic Epitopes on Type XVII Collagen Cause Skin Fragility in a Complement-Dependent and -Independent Manner

Natsuga

Nishie

Shinkuma

et al. 2012

The Journal of Immunology

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In bullous pemphigoid (BP), the most prevalent autoimmune blistering disease, type XVII collagen (COL17) is targeted by circulating autoantibodies. BP is thought to be an autoantibody-mediated complement-fixing blistering disease, and a juxtamembranous noncollagenous 16A (NC16A) domain spanning Glu490 to Arg566 was proved to be the main pathogenic region on COL17, although precise pathogenic epitopes within NC16A have not been elucidated. In this study, we showed that injection of rabbit IgG Abs targeting Asp522 to Gln545 induced skin fragility associated with in vivo deposition of IgG and complement in neonatal COL17-humanized mice. Notably, immunoadsorption of rabbit anti-NC16A IgG Ab with this epitope (Asp522 to Gln545) or the anti-NC16A IgG administered together with the peptides of this epitope as a decoy ameliorated skin fragility in the injected neonatal COL17-humanized mice compared with the anti-NC16A IgG alone even though all of the mice showed both IgG and complement deposition. These results led us to investigate an additional, complement-independent mechanism of skin fragility in the mice injected with anti-COL17 Abs. The rabbit anti-NC16A IgG depleted the expression of COL17 in cultured normal human keratinocytes, whereas immunoadsorption of the same IgG with this epitope significantly suppressed the depletion effect. Moreover, passive transfer of F(ab′)2 fragments of the human BP or rabbit IgG Abs against COL17 demonstrated skin fragility in neonatal COL17-humanized mice. In summary, this study reveals the importance of Abs directed against distinct epitopes on COL17, which induce skin fragility in complement-dependent as well as complement-independent ways.

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Bullous pemphigoid induced by IgG targeting type XVII collagen non‐NC16A/NC15A extracellular domains is driven by Fc gamma receptor‐ and complement‐mediated effector mechanisms and is ameliorated by neonatal Fc receptor blockade

Pigors,

Patzelt,

Reichhelm

et al. 2023

The Journal of Pathology

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Bullous pemphigoid (BP) is an autoimmune blistering disease characterized by autoantibodies targeting type XVII collagen (Col17) with the noncollagenous 16A (NC16A) ectodomain representing the immunodominant site. The role of additional extracellular targets of Col17 outside NC16A has not been unequivocally demonstrated. In this study, we showed that Col17 ectodomain‐reactive patient sera depleted in NC16A IgG induced dermal–epidermal separation in a cryosection model indicating the pathogenic potential of anti‐Col17 non‐NC16A extracellular IgG. Moreover, injection of IgG targeting the murine Col17 NC14–1 domains (downstream of NC15A, the murine homologue of human NC16A) into C57BL/6J mice resulted in erythematous skin lesions and erosions. Clinical findings were accompanied by IgG/C3 deposits along the basement membrane and subepidermal blistering with inflammatory infiltrates. Disease development was significantly reduced in either Fc‐gamma receptor (FcγR)‐ or complement‐5a receptor‐1 (C5aR1)‐deficient mice. Inhibition of the neonatal FcR (FcRn), an atypical FcγR regulating IgG homeostasis, with the murine Fc fragment IgG2c‐ABDEG, a derivative of efgartigimod, reduced anti‐NC14–1 IgG levels, resulting in ameliorated skin inflammation compared with isotype‐treated controls. These data demonstrate that the pathogenic effects of IgG targeting the Col17 domain outside human NC16A/murine NC15A are partly attributable to antibody‐mediated FcγR‐ and C5aR1 effector mechanisms while pharmacological inhibition of the FcRn represents a promising treatment for BP. The mouse model of BP will be instrumental in further investigating the role of Col17 non‐NC16A/NC15A extracellular epitopes and validating new therapies for this disease. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

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Human IgG1 Monoclonal Antibody against Human Collagen 17 Noncollagenous 16A Domain Induces Blisters via Complement Activation in Experimental Bullous Pemphigoid Model

Abstract: Material

Cited by 55 publications

References 58 publications

Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Antibodies to Pathogenic Epitopes on Type XVII Collagen Cause Skin Fragility in a Complement-Dependent and -Independent Manner

Bullous pemphigoid induced by IgG targeting type XVII collagen non‐NC16A/NC15A extracellular domains is driven by Fc gamma receptor‐ and complement‐mediated effector mechanisms and is ameliorated by neonatal Fc receptor blockade

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