title: Monoclonal antibodies against Gal alpha 1-4Gal beta 1-4Glc (Pk, CD77) produced with a synthetic glycoconjugate as immunogen: reactivity with carbohydrates, with fresh frozen human tissues and hematopoietic tumors.
abstract: The purpose of this study was to isolate mouse monoclonal antibodies (MAbs) recognizing Gal alpha 1-4Gal beta 1-4Glc (Pk, CD77), which has been described as a Burkitt's-lymphoma-associated antigen. Three IgGI MAbs reactive with Pk were developed using a synthetic glycoconjugate as immunogen and a filter immunoplaque screening assay. One MAb (PK67) was characterized by immuno-thin-layer chromatography, ELISA and competition assays using neutral glycolipids from various sources and a variety of carbohydrates and glycoproteins. Epitope analysis showed that all 3 carbohydrates moieties are required in PK67 binding. No cross-reactivity was observed with closely related carbohydrate structures, with the exception of Gal alpha 1-4Gal beta 1-4GlcNAc. However, the reactivity with this structure was several orders of magnitude lower than with Pk. Immunohistochemical analysis of fresh-frozen tissue specimens showed that Gb3 is widely expressed: prominent staining was observed in many normal tissues, e.g., kidney and gastric tissue and on capillary endothelial cells. In lymph nodes, very weak staining of a few B cells was observed. Flow cytometric analysis of hematopoietic neoplasms showed Gb3 expression on B-cell neoplasms, particularly those with committed B-cell phenotype indicating that, in addition to earlier reports of Gb3 expression on Burkitt's lymphoma, Gb3 is present on other committed B-cell neoplasms.
authors.affiliation: Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
authors.forename: E
authors.initials: E
authors.lastname: Oosterwijk
authors.forename: A
authors.initials: A
authors.lastname: Kalisiak
authors.forename: J C
authors.initials: JC
authors.lastname: Wakka
authors.forename: D A
authors.initials: DA
authors.lastname: Scheinberg
authors.forename: L J
authors.initials: LJ
authors.lastname: Old
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000032:analysis*; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006017:Glycolipids / Q000276:immunology*; D006023:Glycoproteins / Q000138:chemical synthesis / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D007938:Leukemia / Q000473:pathology*; D008223:Lymphoma / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000473:pathology*; D009928:Organ Specificity; D012016:Reference Values; D014281:Trihexosylceramides / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D002241:Carbohydrates; D006017:Glycolipids; D006023:Glycoproteins; D014281:Trihexosylceramides; C018549:globotriaosylceramide