Characterization of rat monoclonal antibodies against human beta-defensin-2.
Defensins are a family of cationic antimicrobial peptides that participate in host defense. Human beta-defensin (hBD)-2 has a potent bactericidal activity against a wide spectrum of microorganisms. Because human gingival epithelium is constantly exposed to a variety of microbial challenges, it is considered that hBD-2 has an important role in the protective mechanisms against oral bacterial infection. However, little is known about the production of hBD-2 in tissues of the oral cavity. Six rat monoclonal antibodies (MAbs) raised against chemically synthesized hBD-2 have been characterized. Rat MAbs were specific for the conformational epitopes on hBD-2, but not to hBD-1. To identify the epitope on hBD-2, a series of six overlapping peptides covering the hBD-2 whole sequence were synthesized and the immunoreactivities of six MAbs were examined. The FCPRRYK domain in hBD-2 was recognized by all six MAbs and suggested to be an epitope region. By immunocytochemistry, hBD-2 was localized focally in the epidermis of the human gingival tissue using the MAbs. The MAbs specifically recognized against hBD-2 will be a useful tool to study the functional role of antimicrobial agents and an important asset in the imaging of oral infection processes.
Kurosawa
Shigeharu
S
Department of Oral Diagnostics, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
Ohta
Mitsuhiro
M
Hayakawa
Mitsuo
M
Kamino
Yoshikazu
Y
Abiko
Yoshimitsu
Y
Sasahara
Hiroshige
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D004817:Epidermis / Q000378:metabolism; D018604:Epitope Mapping; D000939:Epitopes; D005881:Gingiva / Q000276:immunology / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D051381:Rats; D017386:Sequence Homology, Amino Acid; D023083:beta-Defensins / Q000737:chemistry* / Q000276:immunology*
D000911:Antibodies, Monoclonal; C106641:DEFB4A protein, human; D000939:Epitopes; D010455:Peptides; D023083:beta-Defensins