title: A panel of monoclonal antibodies recognizing GPI-anchored ADP-ribosyltransferase ART4, the carrier of the Dombrock blood group antigens.
abstract: ART4 (CD297) is a member of the family of toxin-related ADP-ribosyltransferases (ARTs) and is the carrier of the Dombrock blood group alloantigens (Do). Two mouse monoclonal antibodies (MIMA-52 and MIMA-53), and two rat monoclonal antibodies (N0NI-B4 and NONI-B63) were obtained following immunization of mice with human Do/ART4-transfected cells and of rats with human Do/ART4 cDNA, respectively. All four mAbs recognize Do/ART4-transfected Jurkat cells but not untransfected cells by FACS analysis. Staining of Do/ART4-transfected cells by these mAbs was reduced following treatment of cells with PI-PLC, confirming that Do/ART4 is anchored in the cell membrane by linkage to glycosylphosphatidylinositol as predicted from its amino acid sequence. The four mAbs did not react with Gy(a-) (Dombrock null) erythrocytes but agglutinated other red blood cells. By flow cytometric analysis, all mAbs reacted prominently with erythrocytes, and weakly with peripheral blood monocytes and splenic macrophages, but not with B-lymphocytes or T-lymphocytes. The mAbs reacted weakly also with human umbilical vein endothelial cells and the basophilic leukemia KU-812. Immunohistology revealed staining of epithelia and endothelia on sections of tonsils. In FACS analyses NONI-B4 competed with MIMA-52 for binding to Do/ART4-transfected cells and erythrocytes, whereas NONI-B63 competed with MIMA-53. Neither of the mAbs reacted with mouse ART4-transfected cells, but NONI-B63 and MIMA-53 did react with a mouse/human ART4 chimera, indicating that the epitope recognized by these mAbs lies in the C-terminal half of the protein.
authors.affiliation: Institute of Immunology, University Hospital, Hamburg, Germany.
authors.forename: Ines
authors.initials: I
authors.lastname: Parusel
authors.forename: Sarah
authors.initials: S
authors.lastname: Kahl
authors.forename: Fenja
authors.initials: F
authors.lastname: Braasch
authors.forename: Gustavo
authors.initials: G
authors.lastname: Glowacki
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Halverson
authors.forename: Marion E
authors.initials: ME
authors.lastname: Reid
authors.forename: Alissa
authors.initials: A
authors.lastname: Schawalder
authors.forename: Erika
authors.initials: E
authors.lastname: Ortolan
authors.forename: Ada
authors.initials: A
authors.lastname: Funaro
authors.forename: Fabio
authors.initials: F
authors.lastname: Malavasi
authors.forename: Debbie
authors.initials: D
authors.lastname: Hardie
authors.forename: Sapna
authors.initials: S
authors.lastname: Halder
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Buckley
authors.forename: Friedrich
authors.initials: F
authors.lastname: Haag
authors.forename: Friedrich
authors.initials: F
authors.lastname: Koch-Nolte
mesh_terms: D036002:ADP Ribose Transferases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D018797:Cell Cycle Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D003429:Cross Reactions; D004730:Endothelium, Vascular / Q000276:immunology; D004912:Erythrocytes / Q000276:immunology; D058851:GPI-Linked Proteins; D006801:Humans; D008264:Macrophages / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009000:Monocytes / Q000276:immunology; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014066:Palatine Tonsil / Q000378:metabolism; D051381:Rats; D016415:Sequence Alignment; D013154:Spleen / Q000276:immunology; D014471:Umbilical Veins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D018797:Cell Cycle Proteins; C102519:GML protein, human; D058851:GPI-Linked Proteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D036002:ADP Ribose Transferases; C105234:ART4 protein, human