title: Unique mouse monoclonal antibodies reactive with maturation-related epitopes on type VII collagen.
abstract: In this study, we generated a new set of monoclonal antibodies (mAbs) to bovine and human type VII collagen (COL7) by immunizing mice with bovine cornea-derived basement membrane zone (BMZ) fraction. The four mAbs, tentatively named as COL7-like mAbs, showed speckled subepidermal staining in addition to linear BMZ staining of normal human skin and bovine cornea, a characteristic immunofluorescence feature of COL7, but showed no reactivity with COL7 by in vitro biochemical analyses. Taking advantage of the phenomenon that COL7-like mAbs did not react with mouse BMZ, we compared immunofluorescence reactivity between wild-type and COL7-rescued humanized mice and found that COL7-like mAbs reacted with BMZ of COL7-rescued humanized mice. In ELISAs, COL7-like mAbs reacted with intact triple-helical mammalian recombinant protein (RP) of COL7 but not with bacterial RP. Furthermore, COL7-like mAbs did not react with COL7 within either cultured DJM-1 cells or basal cells of skin of a bullous dermolysis of the newborn patient. These results confirmed that COL7-like mAbs reacted with human and bovine COL7. The epitopes for COL7-like mAbs were considered to be present only on mature COL7 after secretion from keratinocytes and deposition to BMZ and to be easily destroyed during immunoblotting procedure. Additional studies indicated association of the speckled subepidermal staining with both type IV collagen and elastin. These unique anti-COL7 mAbs should be useful in studies of both normal and diseased conditions, particularly dystrophic epidermolysis bullosa, which produces only immature COL7.
authors.affiliation: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, Kurume, Japan.|Department of Oral and Maxillofacial Surgery, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
authors.forename: Taihei
authors.initials: T
authors.lastname: Hayakawa
authors.affiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Hirako
authors.affiliation: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, Kurume, Japan.
authors.forename: Kwesi
authors.initials: K
authors.lastname: Teye
authors.affiliation: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, Kurume, Japan.
authors.forename: Atsunari
authors.initials: A
authors.lastname: Tsuchisaka
authors.affiliation: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, Kurume, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Koga
authors.affiliation: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, Kurume, Japan.
authors.forename: Norito
authors.initials: N
authors.lastname: Ishii
authors.affiliation: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, Kurume, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Karashima
authors.affiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Minori
authors.initials: M
authors.lastname: Kaneda
authors.affiliation: Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Oyu
authors.affiliation: Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Chiharu
authors.initials: C
authors.lastname: Tateishi
authors.affiliation: Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Sugawara
authors.affiliation: Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Ayano
authors.initials: A
authors.lastname: Yonamine
authors.affiliation: Department of Dermatology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Shinkuma
authors.affiliation: Department of Dermatology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimizu
authors.affiliation: Department of Oral and Maxillofacial Surgery, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Fukano
authors.affiliation: Department of Oral and Maxillofacial Surgery, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Shimozato
authors.affiliation: Department of Dermatology, Stanford University School of Medicine, Center for Clinical Sciences Research, Stanford, CA, USA.
authors.forename: Ngon T
authors.initials: NT
authors.lastname: Nguyen
authors.affiliation: Department of Dermatology, Stanford University School of Medicine, Center for Clinical Sciences Research, Stanford, CA, USA.|Department of Dermatology, Veterans Administration Hospital, Palo Alto Medical Center, Palo Alto, CA, USA.
authors.forename: M Peter
authors.initials: MP
authors.lastname: Marinkovich
authors.affiliation: Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Tsuruta
authors.affiliation: Department of Dermatology, Kurume University School of Medicine, Kurume University Institute of Cutaneous Cell Biology, Kurume, Japan.
authors.forename: Takashi
authors.identifier: 0000-0002-0144-3255
authors.initials: T
authors.lastname: Hashimoto
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001485:Basement Membrane / Q000378:metabolism*; D002417:Cattle; D024143:Collagen Type VII / Q000276:immunology* / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice
chemical_list: D000911:Antibodies, Monoclonal; D024143:Collagen Type VII
keywords: basement membrane zone; dystrophic epidermolysis bullosa; epidermolysis bullosa acquisita; keratinocyte; type VII collagen