Carbohydrate antigen profiles on human erythroleukemia cell lines HEL and K562 (original) (raw)
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Carbohydrate antigen profiles of human erythroleukemia cell lines HEL and K562
Blood, 1983
The expression of major carbohydrate antigens carried by polylactosaminyl chains in human erythroleukemia cell lines, K562 and HEL, was investigated by applying monoclonal antibodies recognizing specific carbohydrate determinants. The two cell lines showed common differences in their glycolipid compositions: (1) the presence of significant amounts of ganglio-series glycolipids, which are absent in normal erythrocytes; and (2) a remarkable reduction in the amount of globo-series glycolipids, which are the major glycolipids in normal human erythrocytes. A variety of differences were also detected in the carbohydrate antigens carried by lacto-series glycolipids and glycoproteins having related carbohydrate chains. K562 cells were i+H- X+, with a minor population of I+ cells. HEL cells were I-i+H+X-. The presence of the I+ population in K562 cells is particularly noteworthy, since I-antigen is characteristic of adult mature erythrocytes and is absent in most human leukemic cell lines. S...
International Journal of Cancer, 1984
Two monoclonal antibodies (KH-I and KH-2) against a transplanted fibrosarcoma (KMT-17) in WKA rab were produced by fusing a mouse myeloma (Pl-X63-Ag8.653) with spleen cells from syngeneic rats hyperimmunized with KMT-17. Both antibodies showed complement-dependent cytotoxicity against KMT-17. By absorption of cytotoxicity. KH-I reacted with homologous tumor, other syngeneic fibrosarcomas (KMT-80 and KMT-75). and lung and kidney from normal rats. However, KH-2 reacted with many kinds of tumors and various normal tissues. Antigen specificity was tested by complement fixation and/or solidphase radioimmunoassay using glycolipids isolated from KMT-17 cells and authentic glycolipids. KH-I reacted with globotriglycosyl ceramide which war not detected on
Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1985
Six monoclonal antibodies with known specificities for the carbohydrate antigens i, X or Y, and seven anti-myeloid antibodies (determinants unknown) selected for their differing reaction patterns with human leucocytes were tested in chromatogram binding assays for reactions with myeloid cell glycolipids derived from normal human granulocytes and chronic myelogenous leukemia cells. Antigenicities were found exclusively on minor glycolipids which were barely or not at all detectable with orcinol-sulphuric acid stain. Among these, a neutral glycusphingolipid bound the anti-i antibody Den and chromatographed as the ceramide octasaccharide, Gait1-, 4GicNAc~I ~ 3Gait1 ~ 4GIcNAcfll ~ 3Gait1 ~ 4GlcNAc~1-, 3Gait1-> 4GIc-Cer. Several species of neutral glycosphingolipids with six to more than ten monosaccharides were detected which carry the X antigen and others the Y antigen: Galfll-, 4(Fucal ~ 3)GIcNAc and Fucal ~ 2Galfll-*4(Fucal ~3)GIcNAc, respectively. In addition, three new types of carbohydrate specificities were detected among the myeloid cell glycolipids. Two were associated with neutral glycolipids: the first, recognised by anti-myeloid antibodies VIM-1 and VIM-10, was expressed on a distinct set of glycolipids with six or more monosaccharides, and the second, recognized by VIM-8, was expressed on glycolipids with more than ten monosaccharides. The third specificity, recognised by the anti-myeloid antibody VIM-2, was expressed on slow migrating sialoglycolipids with backbone structures of the poly-Nacetyilactosamine type that are susceptible to degradation with endo-fl-galactosidase. Thus, we conclude that the i and Y antigens occur among the glycolipids of normal myeloid and chronic myelogenous leukemia cells and that a high proportion of hybridoma antibodies raised against differentiation antigens of myeloid cells are directed at carbohydrate structures.
Glycosphingolipids of K562 cells: A chemical and immunological analysis
International Journal of Cancer, 1981
The K562 cell line, which was established from a patient with chronic myeloid leukemia in blast crisis, was thought to be myeloid, but recent data indicate that it is an undifferentiated erythroid cell line. We have found that the glycosphingolipid content of these cells differs considerably from that of mature erythrocytes. Globotetraosylceramide, the most abundant glycolipid of mature red cells, was not detected in K562 cells, and neither was globotriaosylceramide. The predominant neutral glycolipids of K562 cells are monohexaosylceramides, which are a mixture of glucosyl-and galactosylceramides, and lactotriaosyl-and lactoneotetraosylceramides were also detected. Secondly, gangliosides which contain Kacetylgalactosamine were much more abundant than those containing N-acetylglucosamine in K562 cells, in contrast to erythrocytes. The most abundant ganglioside of K562 cells, G, , , is present in trace quantities in erythrocytes. A third major difference between these two cells lies in their relative proportions of neutral glycolipids and gangliosides. The molar ratio of neutral glycolipidslgangliosides is approximately IS:I in erythrocytes and I:I in K562 cells. These striking differences between K562 cells and mature erythrocytes indicate that glycolipids may be useful cell surface markers of normal erythrocyte differentiation, and of erythroleukemias.
Pure and Applied Chemistry, 1977
Four sets of glycolipid variante of human A and 0 erythrocytes were isolated and their structures were determined. The glycolipids carrying A-determinants were termed Aa, Ab, AC and Ad according to the complexity of the carrier carbohydrate chain. in that order. Glycolipids with H-activity were termed H1, ~. H3 and H4 according to the order of their complexity of the carrier carbohydrate chain. It is suggested that the conversion of complex H variante (H3/or H4) to complex A variante (AC/or Ad) was significantly lower in A2 erythrocytes. In fetal/er newborn erythrocytes the highly complex variante (Ac, Ad, H3 and H4) were significantly lower • than adult erythrocytes. These and other studies suggest that step-by-step elongation and arborization of complex glycolipids such as Ac and H3 chains may take place during the process of development. A remarkable difference between the reaction of glycolipids in human intestinal tumors and the reaction of normal intestinal mucosa was demonstrated by the antibody which was directed against core structure of carrier carbohydrate chain, that is GlcNAcßl~3Galßl~4Glc~er. This structure must be present in greater quantity in ~umor tissue than in normal mucosa.
Cancer Research, 1985
Three mouse monoclonal antibodies to distinct cell surface antigens were derived from immunizations with cells of Tera-1, a human teratocarcinoma cell line, and a membrane preparation of placenta! tissue. The distribution of the antigens on 165 cultured lines of various human tumors and normal cells was determined by mixed hemadsorption assays and on fresh tissues by immunofluorescence staining. K4 antigen is expressed on cell lines derived from teratocarcinomas but not on any other cultured cell tested. Normal adult colonie epithelium, some fetal tissues, and specimens of testicular teratocarcinoma were also K4 posi tive. K21 antigen was detected on teratocarcinoma cell lines and, at more than 100-fold lower levels, on cultures of normal and malignant kidney epithelium but not on other cultured cells. K21 expression in normal tissues is restricted to the epithelium of fetal intestine and bronchus. Other fetal tissues and all adult normal tissues tested lacked K21. A subset of teratocarcinoma specimens (5 of 8) was reactive with antibody K21. P12 antigen is represented on a wide range of cell lines and tissues, including a subset of teratocarcinomas. AbK4, AbK21, and AbP12 react with carbohydrate sequences present on high-molecular-weight glycoproteins. AbK21 and AbP12 recognize the lactc~A/-tetraose and lacto-A/-fucopentaose III (X-hapten) structures, respectively, whereas AbK4 reacts with a neuraminidase-sensitive determi nant.
Bioscience Reports, 1983
The thin-layer-chromatography immunostaining procedure was applied to human erythrocyte glycolipids using monoclonal anti-i and anti-I antibodies which are directed against epitopes on linear and branched carbohydrate chains of the neolacto (poly-N-acetyl-lactosamine) series. An examination of native and mild-acid-treated glycolipids from normal adult (Iadult antigen type), neonatal (icord), and I-antigen-deficient adult (iadult) erythrocytes enabled certain structural inferences to be made as follows: (a) cells of both I and i phenotypes contain a multiplicity of glycolipids of the neolacto series whose backbones consist of 8 or more sugar residues; (b) the octasaccharide backbones are predominantly linear in cells of i phenotype and branched in those of I type; and (c) more complex glycolipids having decasaccharide and larger backbones with both linear and branched sequences occur in erythrocytes of both phenotypes, 0