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Unique mouse monoclonal antibodies reactive with maturation‐related epitopes on type VII collagen
Author(s) -
Hayakawa Taihei,
Hirako Yoshiaki,
Teye Kwesi,
Tsuchisaka Atsunari,
Koga Hiroshi,
Ishii Norito,
Karashima Tadashi,
Kaneda Minori,
Oyu Yuka,
Tateishi Chiharu,
Sugawara Koji,
Yonamine Ayano,
Shinkuma Satoru,
Shimizu Hiroshi,
Fukano Hideo,
Shimozato Kazuo,
Nguyen Ngon T.,
Marinkovich M. Peter,
Tsuruta Daisuke,
Hashimoto Takashi
Publication year - 2017
Publication title -
experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.108
H-Index - 96
eISSN - 1600-0625
pISSN - 0906-6705
DOI - 10.1111/exd.13306
Subject(s) - epitope , monoclonal antibody , chemistry , antibody , microbiology and biotechnology , immunology , biology
In this study, we generated a new set of monoclonal antibodies ( mA bs) to bovine and human type VII collagen ( COL 7) by immunizing mice with bovine cornea‐derived basement membrane zone ( BMZ ) fraction. The four mA bs, tentatively named as COL 7‐like mA bs, showed speckled subepidermal staining in addition to linear BMZ staining of normal human skin and bovine cornea, a characteristic immunofluorescence feature of COL 7, but showed no reactivity with COL 7 by in vitro biochemical analyses. Taking advantage of the phenomenon that COL 7‐like mA bs did not react with mouse BMZ , we compared immunofluorescence reactivity between wild‐type and COL 7‐rescued humanized mice and found that COL 7‐like mA bs reacted with BMZ of COL 7‐rescued humanized mice. In ELISA s, COL 7‐like mA bs reacted with intact triple‐helical mammalian recombinant protein (RP) of COL 7 but not with bacterial RP. Furthermore, COL 7‐like mA bs did not react with COL 7 within either cultured DJM ‐1 cells or basal cells of skin of a bullous dermolysis of the newborn patient. These results confirmed that COL 7‐like mA bs reacted with human and bovine COL 7. The epitopes for COL 7‐like mA bs were considered to be present only on mature COL 7 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‐ COL 7 mA bs should be useful in studies of both normal and diseased conditions, particularly dystrophic epidermolysis bullosa, which produces only immature COL 7.