CD88 antibodies specifically bind to C5aR on dermal CD117+ and CD14+ cells and react with a desmosomal antigen in human skin.
The expression of the C5aR (CD88) on human epidermal and dermal cells was studied with five anti-C5aR mAb directed to the N-terminal domain of the receptor. All mAb bound to suspended dermal CD117+ mast cells and to dermal CD14+ cells. The binding to CD14+ and CD117+ cells could be blocked by rC5a and by peptide EX-1 representing amino acid residues 1-31 of the C5aR. In acetone-fixed frozen or in paraformaldehyde-fixed, paraffin-embedded tissue, we detected a binding of the Abs to dermal perivascular cells and, additionally, to keratinocytes and dermal epithelial cells that could be blocked by EX-1. Immunoelectromicroscopy revealed a binding of anti-C5aR mAb to desmosomal regions in human epidermis. However, the following results indicate that CD88 mAb cross-react with epithelium in a specific way: 1) the binding to suspended epidermal cells and to the epidermal cell line HaCat could be blocked by EX-1 but not by rC5a; 2) FITC-labeled C5a bound to CD117+ and to CD14+ cells but not to epidermal cells; 3) C5a led to transient calcium fluxes in CD14+ and CD117+ dermal but not in epidermal cells; 4) C5aR mRNA was detectable by reverse transcription PCR in granulocytes but not in keratinocytes or in HaCat. Our results show that CD88 mAb are good tools for the investigation of the C5aR on hemopoietic cells. Results with epithelial cells should be considered with caution, as the binding of CD88 mAb that were raised to a synthetic peptide sequence may be due to a cross-reactivity.
Werfel
T
T
Department of Dermatology, Hannover Medical School, Germany.
Zwirner
J
J
Oppermann
M
M
Sieber
A
A
Begemann
G
G
Drommer
W
W
Kapp
A
A
Götze
O
O
D000911:Antibodies, Monoclonal / Q000276:immunology*; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003896:Desmosomes / Q000276:immunology*; D000078404:Epidermal Cells; D004817:Epidermis / Q000276:immunology; D006801:Humans; D015603:Keratinocytes / Q000276:immunology; D018950:Lipopolysaccharide Receptors / Q000032:analysis*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis*; D012333:RNA, Messenger / Q000096:biosynthesis; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D012867:Skin / Q000166:cytology / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018950:Lipopolysaccharide Receptors; D012333:RNA, Messenger; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement; D019009:Proto-Oncogene Proteins c-kit