Monoclonal antibodies produced by immunization with neoglycoproteins containing galα1‐4galβ1‐4glcβ‐O and galα1‐4galβ1‐4glcnacβ‐O residues: Useful immunochemical and cytochemical reagents for blood group p antigens and a differentiation marker in burkitt lymphoma and other B‐cell malignancies

Abstract

Several monoclonal antibodies (MAbs) directed to blood group P1 (Gal alpha 1-4Gal beta 1-4GlcNAc beta-O) and Pk (Gal alpha 1-4Gal beta 1-4Glc beta-O) determinants were produced with high efficiency by using synthetic neoglycoproteins as immunogens. The specificity of IgM and IgG1 MAbs was characterized by binding to defined oligosaccharides and glycoconjugates. Antibodies that bound equally well to P1 and Pk determinants and to Gal alpha 1-4Gal beta 1-O-derivatives were obtained, together with others that showed selective recognition of the entire trisaccharide chain. Selected antibodies were shown to be useful as reagents for detection of the blood group P antigens in glycolipid extracts of erythrocytes and on the surface of erythrocytes of different P phenotypes, demonstrated both by radioimmune assays and hemagglutination. Six IgM MAbs directed to the Pk determinant bound selectively to Burkitt lymphoma cells and 2 of these antibodies (424/3D9 and 424/6A2) could be used as auxiliary reagents in immunofluorescence for diagnosis and classification of B-cell lymphomas and leukemias using flow cytometric analysis. Eight normal individuals and 37 patients with lymphoma or leukemia were studied. Tumor cells of 2/2 patients with "Burkitt-like" lymphoma, 1 patient with centroblastic lymphoma and 2 patients with acute leukemia were strongly stained for the Pk antigen. The staining patterns for differentiation markers classified the tumor cells to a developmental stage closely related to the Burkitt cell type.

Lipooligosaccharide P k (Galα1-4Galβ1-4Glc) Epitope of Moraxella catarrhalis Is a Factor in Resistance to Bactericidal Activity Mediated by Normal Human Serum

“…These data suggest that resistance to serum complement-mediated killing may be related to the presence of the two terminal galactose residues on the LOS. These residues form part of a structure that has been shown to be immunochemically identical to the P k (Gal␣1-4Gal␤1-4Glc) antigen found on a number of human cells including erythrocytes, as well as gastrointestinal, ureteral, and bladder epithelial cells (7,29,35). It would appear that the Moraxella Gal␣1-4Gal␤1-4Glc structure may act as a human self-antigen and that antibodies to it are not present in normal human serum.…”

Glycosphingolipid expression in acute nonlymphocytic leukemia: common expression of shiga toxin and parvovirus B19 receptors on early myeloblasts

“…27,32,39 Finally, Gb 4 has been reported on the membranes of some clinical ANLL samples by immunofluorescence microscopy and flow cytometry. 53,90 In contrast, Kyogashima et al 83 failed to observe either Gb 3 or Gb 4 in 3 of 3 ANLLs, including 2 cases of acute monocytic leukemia, which might be expected to express globo GSLs. 24,32 Overall, 17 of 20, or 85%, of all clinical ANLL samples published to date have been shown to express globo GSLs, suggesting that globo GSLs are commonly expressed in ANLL.…”

“…24,25,37,38 To screen for Gb 3 and Gb 4 , samples were immunostained with MoAbs Pk002 and MC631, respectively ( Figure 2D,E). 25,53,54 MoAb Pk002 binding to Gb 3 (R f 0.45) was observed in all 3 *GSLs were identified by the size of the oligosaccharide moiety, relative mobility (Rf), and immunostaining characteristics (Figure 2). †Percent distribution of major neutral GSL species as determined by scanning densitometry of DPA-stained HPTLC plates.…”

“…53 MoAb MC631 was purchased as a hybridoma supernatant from the Developmental Studies Hybridoma Bank maintained by the Department of Biology at the Univiversity of Iowa, Iowa City, under contract no. N01-HD-2-3144.…”

Structural Relationships and Sialylation among Meningococcal L1, L8, and L3,7 Lipooligosaccharide Serotypes

“…The L8 mAb, 2-1-L8, has been characterized extensively; it binds the lactosyl ␣-oligosaccharide of 3.6-kDa neisserial LOS (2,21,23,29). mAbs that bind the P k (Gal␣134Gal␤134Glc3 ceramide) (22,30), P 1 (Gal␣134Gal␤134-GlcNAc-R), and Gal␣134Gal-R human GSL were bought from Mono-Carb (Accurate Chemicals, Westbury, NY).…”