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Biomedical subjects

G C Hansson

Publications and source records attributed to G C Hansson.

At least 91 records · Page 5Linked to original sources
PubMed1988

The glycosphingolipid composition of the placenta of a blood group P fetus delivered by a blood group Pk1 woman and analysis of the anti-globoside antibodies found in maternal serum.

To further define the molecules that may mediate spontaneous abortion due to maternal-fetal blood group incompatibility within the P blood group system, we have examined the fine specificities of maternal antibodies and the glycolipid antigens from the placenta of a P infant born to a Pk1 mother. Maternal antibodies obtained during therapeutic plasmapheresis were analyzed to determine their reactivities with placental glycolipid extracts on thin-layer plates. Second antibodies specific for IgM, IgG, and IgA revealed immunoglobulins of all of these classes strongly reactive with one major placental glycolipid that comigrates with globoside. GC/MS analysis confirmed that the major P-active pentaglycosylceramide of placenta has the same structure as that previously shown for the P antigen of red blood cells: GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc-Cer. Serum antibodies partially purified by affinity chromatography on globoside-octyl-Sepharose specifically recognize glycolipids that contain terminal GalNAc beta 1-3Gal . . . residues and also recognize the same sequence as an internal determinant in some, but not all, glycolipids with extended globoside core regions. Thus, in the blood group P incompatible fetus, the major P antigen present in placenta has the same carbohydrate structure as the P antigen present in fetal and adult erythrocytes and might be a target for the maternal immune system.

Adult↗
PubMed1988

Intestinal-type glycosphingolipids in urine from patients with enterocutaneous urinary diversions.

Ureteroenterocutaneous stoma is a surgical procedure for urinary diversion by which an intestinal segment is used as a conduit or reservoir for the urine. The intestinal mucosa continues to produce a mucous gel containing desquamated epithelial cells. This mucous gel is easily obtainable from the urine. Glycosphingolipids were isolated from such gels and analyzed with immunological methods using a thin-layer chromatography overlay assay and mass spectrometry. The glycosphingolipids obtained mainly belong to the lacto-type I series comprising ABH, Lea, and Leb determinants. These observations support the intestinal origin of the glycosphingolipids. A regional distribution of the ABH and Lewis antigens, which is in accordance with previous immunohistochemical findings, was observed.

Chromatography, Thin Layer↗
PubMed1988

Mucins obtained from patients with enterocutaneous urinary diversions.

The ureteroenterocutaneostomy is a surgical procedure for urinary diversion by which an intestinal segment is used as a conduit or a reservoir for urine. Histological studies on colonic segments have shown a well preserved mucosal surface with numerous goblet cells and urine from these patients contains a mucus-like sediment. The present study demonstrates that such gels are composed of mucins occurring predominantly as a glycoprotein complex which is 'insoluble' in 6 M guanidinium chloride. The complex was brought into solution by reduction of disulphide bonds and the fragments (subunits) obtained behaved as typical mucins when subjected to density-gradient centrifugation in CsCl/4 M guanidinium chloride. High-Mr glycopeptides obtained after trypsin digestion of subunits expressed the ABH and Lewis antigens, in accordance with the patients' blood-group phenotype and with the regional distribution known from previous immunohistochemical studies on normal colonic mucosa. A heterogeneous population of mucin glycopeptides was revealed by using high-performance ion-exchange chromatography and the major carbohydrate-containing peak did not coincide with that expressing the blood-group activity.

ABO Blood-Group System↗
PubMed1987

Rapid characterization of mucin oligosaccharides from rat small intestine with gas chromatography-mass spectrometry.

Mucin glycopeptides were isolated from rat small intestinal mucosa after reduction/alkylation, trypsin digestion and gel chromatography. The oligosaccharides were released by using alkaline-NaBH4, separated into neutral and acidic species and permethylated. The derivatized mixtures were analysed with fast atom bombardment mass spectrometry and gas chromatography-mass spectrometry using thin film columns. Permethylated neutral oligosaccharides with up to seven sugars could be chromatographed and detected with mass spectrometry. The complex mixture revealed was partly due to the linkage GalNAc being substituted at both position 3 and 6. The approach will be very useful when analysing small amounts of mucins and mucin fragments.

Animals↗
PubMed1987

Developmental changes of gangliosides of the rat stomach. Appearance of a blood group B-active ganglioside.

Rat stomach gangliosides were purified and their distribution in the different tissue compartments was established. Three major monosialogangliosides were found: GM3, GM1, and a ganglioheptaosylceramide carrying a blood group B determinant. This latter structure was characterized by exoglycosidase degradation, immunostaining with a monoclonal anti-blood group B antibody on thin layer chromatogram, permethylation analysis, electron-impact mass spectrometry of the permethylated-reduced and trimethylsilylated molecule, and 1H NMR spectroscopy of the native ganglioside. It was found to be (Formula: see text) i.e. a GM1 structure substituted with the blood group B determinant and was called B-GM1. A similar structure has been previously identified in precancerous rat liver and chemically induced rat hepatoma (Holmes, E. H., and Hakomori, S. (1982) J. Biol. Chem. 257, 7698-7703). Fucosyl-GM1 was also detected as a minor ganglioside in rat gastric mucosa. The ganglioside profile was modified during the postnatal development. The contribution of GM3 and GD3, which accounted for 95% of the ganglioside sialic acid at birth, decreased during the first 3 weeks of life. GM1, fucosyl-GM1, and B-GM1 were not detected at birth. The concentration of the fucogangliosides increased during the 2nd and 3rd weeks after birth, was stable during the 4th week and then decreased, whereas that of GM1 increased steadily between 6 days and 2 months of age. B-GM1, which has been defined as a tumor-associated ganglioside in the rat liver, was found to be a developmentally regulated antigen of the normal rat stomach.

ABO Blood-Group System↗
PubMed1987

The mono- and difucosyl blood group B glycosphingolipids of rat large intestine differ in type of core saccharide.

Two blood group B-active glycosphingolipids were isolated from rat large intestine and characterized by mass spectrometry, proton NMR spectroscopy and methylation analysis. The following structures were concluded: Gal alpha 1----3(Fuc alpha 1----2)Gal beta 1----3GlcNAc beta 1----3Gal beta 1----4Glc beta 1----1Cer and Gal alpha 1----3(Fuc alpha 1----2)Gal beta 1----4(Fuc alpha 1----3)GlcNAc beta 1----3Gal beta 1----4Glc beta 1----1Cer. The two glycolipids thus differ in their core saccharides (type 1 and type 2 chain, respectively) and therefore must have different pathways for biosynthesis.

ABO Blood-Group System↗
PubMed1987

Characterization of neutral blood group B-active glycosphingolipids of rat gastric mucosa. A novel type of blood group active glycosphingolipid based on isogloboside.

The blood group active glycosphingolipids of rat gastric mucosa have been investigated. Only blood group B active structures were found, two of which have been structurally characterized by monoclonal antibodies, mass spectrometry, permethylation analyses, proton NMR spectroscopy, and exoglycosidase digestions. A six-sugar compound based on a gangliotetraosylceramide core was isolated and shown to have the following structure: (Formula: see text). The same compound was recently isolated from rat bone marrow cells and characterized by Taki et al. (Taki, T., Kimura, H., Gasa, S., Nakamura, M., and Matsumoto, M. (1985) J. Biol. Chem. 260, 6219-6225). The possible precursor compounds of this structure, gangliotriaosylceramide and gangliotetraosylceramide, were also found in the gastric mucosa. A seven sugar compound, based on isogloboside, was isolated from the gastric mucosa and shown to have the following structure: (formula; see text) The latter compound is novel and extends the list of different types of core structures found for blood group glycolipids. The epithelial cells of the stomach are unique among the cells lining the gastrointestinal tract in having blood group active glycolipids based on ganglio- and isogloboseries core structures.

ABO Blood-Group System↗
PubMed1987

Structures of blood group glycosphingolipids of human small intestine. A relation between the expression of fucolipids of epithelial cells and the ABO, Le and Se phenotype of the donor.

Small intestinal epithelial cells (enterocytes) were isolated from specimens obtained at operation from four human individuals with different blood group ABO, Lewis, and secretor phenotypes. The non-acid glycolipids were isolated and characterized by thin-layer chromatography, mass spectrometry, and proton NMR spectroscopy and for reactivity with monoclonal antibodies on thin-layer chromatograms. Monohexosylceramides and blood group ABH (type 1 chain) and Lewis glycolipids with 5-7 sugar residues were the major compounds present in all cases, and the expression of the major blood group glycolipids was in agreement with the ABO, Lewis, and secretor phenotype of the individual donors. Small amounts of more complex glycolipids with up to 10 sugar residues were identified by mass spectrometry in all cases. In addition, small amounts of lactotetraosylceramide, a blood group H-active triglycosylceramide with the structure of Fuc alpha 1-2Gal-Hex-Cer (where Fuc is fucose, Hex is hexose, and Cer is ceramide), and dihexosylceramides were identified in some cases. Globotriaosyl- and globotetraosylceramides were absent from the epithelial cells. Small amounts of Leb-active glycolipids in blood group OLe(a+b-), non-secretor and OLe(a-b-), secretor individuals as well as trace amounts of type 2 carbohydrate chain compounds in all individuals were detected by specific monoclonal antibodies.

ABO Blood-Group System↗
PubMed1987

Application of a simple methylation procedure for the analyses of glycosphingolipids.

Acid and neutral glycosphingolipids (0.01-1 mumol) were completely methylated in high yields and with little formation of by-products in 10 min at room temperature, using methyl sulphoxide, methyl iodide, and powdered NaOH. Re-methylation of methylated and LiAlH4-reduced gangliosides gave a new derivative that was useful for the analysis of gangliosides by mass spectrometry.

Animals↗
PubMed1986

Separation and partial sequence analysis of blood group A-active oligosaccharides by affinity chromatography using monoclonal antibodies.

An affinity column containing an anti-blood group A monoclonal antibody coupled to Sepharose beads specifically retards oligosaccharides with the nonreducing trisaccharide sequence GalNAc alpha 1-3(Fuc alpha 1-2)Gal beta 1-R. Three A-active oligosaccharides, A-tetra, A-penta, and A-hepta, elute as retarded peaks, well-separated from unbound sugars. A-hepta, which contains a difucosylated type 1 (Leb) core structure, elutes much later than A-tetra or A-penta and can be completely separated from the latter oligosaccharides by affinity chromatography. The order of elution of the oligosaccharides agrees with their previously determined specific molar activities as inhibitors of quantitative immune precipitation [H.-T. Chen, and E. A. Kabat, (1985) J. Biol. Chem. 260, 13208-13217]. Treatment of A-hepta with Charonia lampas alpha-galactosaminidase abolishes its binding by the anti-A affinity column and converts it to a Leb-active oligosaccharide (lacto-N-difucohexaose I) that is specifically retarded on a second affinity column containing an anti-Leb monoclonal antibody.

ABO Blood-Group System↗
PubMed1986

A novel sulfoglycosphingolipid of mouse small intestine, IV3-sulfogangliotetraosylceramide, demonstrated by negative ion fast atom bombardment mass spectrometry.

In a comparative study of acidic glycosphingolipids in the small intestine of several species it was found that the mouse contained a complex sulfoglycolipid as a major component (Breimer, M.E., Hansson, G.C., Karlsson, K.-A., and Leffler, H. (1983) J. Biochem. (Tokyo) 93, 1473-1485). Fast atom bombardment negative ion mass spectrometry proved the presence and location of the sulfate group and also showed the saccharide sequence and ceramide composition. Combined with NMR spectroscopy of the intact structure and degradative studies the structure was shown to be -O3SO----3Galp beta 1----3GalNAcp beta 1----4Galp beta 1----4Glcp beta 1----1Cer. The sulfoglycolipid was enriched in epithelial cells of mouse small intestine where it constituted at least 90% of the acidic glycolipids and 4-8 mol% of the total glycosphingolipids.

Animals↗
PubMed1986

Two strains of the Madin-Darby canine kidney (MDCK) cell line have distinct glycosphingolipid compositions.

The glycosphingolipids (GSLs) of two sublines of Madin-Darby canine kidney (MDCK) cells, an epithelial cell line, were characterized by t.l.c., antibody overlay and mass spectrometry. The major characteristic which distinguishes the two MDCK cell strains is their trans-epithelial electrical resistance which is typically of the order of 3000 ohm.cm2 for strain I and 100 ohm.cm2 for strain II cells. Strain I and II cells were equally rich in glycolipids, the cellular GSL/phospholipid ratio being 0.04. However, while the phospholipid patterns were identical, the GSLs showed striking differences, and each cell strain expressed appreciable amounts of GSLs that were not found in the other strain. Both cell types possessed neutral GSLs with one, two or three carbohydrate moieties. The monoglycosylceramide accounted for 50% of the total GSLs in each strain. However, while in strain I cells over 90% of this monoglycosylceramide was monoglucosylceramide, in strain II cells over 90% consisted of monogalactosylceramide. In addition, MDCK strain II cells selectively expressed GSLs belonging to the globo series (26% of its neutral GSLs), including globoside and Forssman antigen, a globoside derivative. MDCK strain I cells, on the other hand, expressed another series of GSLs with 4-7 carbohydrate moieties characterized by the common sequence Hex-HexNAc-Hex-Hex-Cer. The presence of two fucosylated GSLs in these series was established. Both MDCK strain I and II cells contained negatively charged GSLs, the major component of which was the ganglioside GM3. MDCK strain II cells in addition expressed sulfatide, the sulfated derivative of galactosylceramide.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Biosynthesis of the cancer-associated sialyl-Lea antigen.

A cancer-associated glycolipid antigen defined by monoclonal antibody 19-9 has the structure NeuAc alpha 2-3Gal Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc beta 1-Cer. We have (formula; see text) studied its biosynthesis by testing the capacity of a crude microsomal fraction of SW 1116 cells to catalyze the addition of fucosyl or sialyl residues from GDP-fucose or CMP-sialic acid to glycolipid or oligosaccharide precursors. When the tetrasaccharide NeuAc alpha 2-3Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc (LSTa) is incubated with GDP-[14C]fucose and SW 1116 microsomes, a 14C-labeled oligosaccharide is formed that can be separated from the incubation mixture on an affinity column containing antibody 19-9 bound to protein A-Sepharose. The product migrates slower than LSTa when analyzed by paper or thin-layer chromatography. After treatment with neuraminidase, it co-migrates with the pentasaccharide Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc (formula; see text) (LNF II) in both chromatographic systems. Similar experiments demonstrate that SW 1116 microsomes catalyze the addition of a sialyl residue to the tetrasaccharide Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc to form LSTa. However, when LNF II is incubated with CMP-[14C]sialic acid and SW 1116 microsomes, no 19-9-active product is detected by affinity chromatography or by paper or thin-layer chromatography. Results using glycolipid precursors are consistent with these findings and also demonstrate the presence of the Lewis fucosyltransferase in SW 1116 cells. Thus, the biosynthesis of the sialyl-Lea antigen proceeds by addition of sialic acid to a type 1 precursor chain by a sialyltransferase, followed by addition of fucose by the Lewis fucosyltransferase.

Antibodies, Monoclonal↗
PubMed1985

Specificity of binding of a strain of uropathogenic Escherichia coli to Gal alpha 1----4Gal-containing glycosphingolipids.

A strain of Escherichia coli originally isolated from urine of a patient with acute pyelonephritis was studied in detail for binding to glycosphingolipids. Bacteria labeled metabolically with [14C]glucose were layered over a glycolipid chromatogram and bound bacteria were detected by autoradiography. The detection was down to a few ng of glycolipid (pmol level) under these assay conditions. At a test level of 500 ng all glycolipids (more than a dozen molecular species analyzed) with Gal alpha 1----4Gal as an internal or terminal part bound the bacteria strongly while glycolipids known to lack this sequence were negative. Conformational analysis using hard sphere calculations including the exo-anomeric effect showed a bend in the saccharide chain at this disaccharide with a largely hydrophobic surface of the convex side, probably being part of the binding epitope. Mixtures of glycolipids isolated from a human ureter scraping and from urinary sediments bound bacteria in the 2- to 7-sugar interval. Thus, this infectious strain of E. coli recognizes glycolipids being present in epithelial cells lining the urinary tract.

ABO Blood-Group System↗
PubMed1985

Carbohydrate-specific adhesion of bacteria to thin-layer chromatograms: a rationalized approach to the study of host cell glycolipid receptors.

Conditions have been adapted for the binding of intact bacteria to glycosphingolipids in a thin-layer chromatogram. Bacteria labeled externally with 125I or metabolically with other isotopes are layered on the plate and after repeated washing the bound bacteria are detected by autoradiography. Using this technique several kinds of bacteria have been shown to adhere to the plate in a carbohydrate-specific way with practically no background binding. Among the advantages of the method is the possible detection of a minor receptor component of a complex mixture extracted from a target cell, facilitating the isolation of the receptor for structural studies. In addition, the multivalent solid-phase presentation of the receptor candidate should also reveal low-affinity binding sites, which may escape detection in traditional inhibition experiments with soluble oligosaccharides.

Adhesiveness↗
PubMed1985

The specific glycosphingolipid composition of human ureteral epithelial cells.

Total non-acid and acid glycolipid fractions were isolated from epithelial cell scrapings and the non-epithelial residue of a human upper ureter. The glycolipid fractions were structurally characterized as total mixtures by thin-layer chromatography, mass spectrometry, and proton NMR spectroscopy. Selected structural information was also obtained on binding of monoclonal antibodies and bacteria to the thin-layer chromatograms. The major epithelial cell glycolipids were Glc beta 1-1ceramide (75%), dihexosylceramide (10%) and NeuAcLacceramide (10%). In addition, 8 minor glycolipids belonging to the blood group P, Lewis and ABO systems were identified. The major glycolipids of the non-epithelial residues were mono- and dihexosylceramides together with globotriaosyl- and globotetraosylceramides. The epithelial mono- and diglycosylceramide compounds had an unusual ceramide composition with mainly C18 and C20 trihydroxy long chain bases in combination with C22-C24 hydroxy fatty acids in contrast to the non-epithelial glycolipids which contained mainly C18 dihydroxy long chain bases in combination with C16-C24 non-hydroxy fatty acids.

Carbohydrates↗
PubMed1984

Lewis blood group antigens defined by monoclonal anti-colon carcinoma antibodies.

Monoclonal antibodies directed against human cancer cells were prepared by the murine hybridoma technique. These antibodies detect Lewis blood group antigens as determined by indirect solid-phase radioimmunoassay, hapten inhibition studies, and chromatogram binding assay. One monoclonal antibody is specific for the Lea terminal carbohydrate of Gal beta 1----3Glc NAc(4----1 alpha Fuc) beta 1----3LacCer. Five monoclonal antibodies react with the Leb terminal carbohydrate sequence of Fuc alpha 1----2Gal beta 1----3GlcNAc(4----1 alpha Fuc) beta 1----3LacCer, and four of these antibodies are highly specific for this glycolipid and do not react with other similar di- and monofucosylated glycolipids. One of the anti-Leb antibodies cross-reacts with blood group H glycolipid and has binding properties similar to those of the previously described antibody NS-10-17 [M. Brockhaus, J. L. Magnani, M. Blaszczyk, Z. Steplewski, H. Koprowski, K.-A. Karlsson, G. Larson, and V. Ginsburg (1981) J. Biol. Chem. 256, 13223-13225]. Two antibodies react with both the Lea and Leb antigens, though both bind preferentially to Leb.

Antibodies, Monoclonal↗