Specificity of human xenoantibodies formed in response to fetal porcine isletlike cell clusters.
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Animals↗
Biomedical subjects
B E Samuelsson
Publications and source records attributed to B E Samuelsson.
Electron ionization-tandem mass spectrometry of glycosphingolipids. Part II. The identification of a carbohydrate sequence corresponding to a novel repetitive blood group A heptaglycosylceramide.
In a previous paper, the presence in human kidney vein tissue of a novel blood group A heptaglycosylceramide based on the type-3 carbohydrate chain GalNAc alpha 1-3(Fuc alpha 1-2)Gal beta 1-3GalNAc alpha 1-3(Fuc alpha 1-2)Gal beta 1-4Glc beta 1-1 Ceramide, was suggested based on thin-layer immunostaining and electron ionization mass spectrometry. Ions corresponding to a structure containing two deoxyhexoses, two hexosamines and three hexoses were identified, but no information was obtained from mass spectrometry concerning the carbohydrate sequence. In the present paper, we report the identification of carbohydrate sequence ions corresponding to a type-3 chain A heptaglycosylceramide by electron ionization-tandem mass spectrometry of a permethylated-reduced glycosphingolipid mixture isolated from human kidney vein tissue. The use of a microchannel-plate-array detector increased the sensitivity for collision-induced dissociation spectra by a factor of at least ten over a conventional electron multiplier.
Blood Group Antigens↗
Expression of glycolipid blood group antigens in single human kidneys: change in antigen expression of rejected ABO incompatible kidney grafts.
Total neutral glycolipid fractions were separated into molecular species on thin-layer chromatography plates and detected by immunostaining with monoclonal anti-blood group antibodies. Blood group A antigens based on type 1, 2, 3 and 4 carbohydrate core saccharides were present in kidneys of A1 and A1B individuals. Blood group A2 individuals expressed only small amounts of A antigen compared to A1 individuals especially of the type 3 and 4 compounds. Kidneys from non-secretor individuals contained less A antigen compared to secretor individuals, and in both groups a variation in the antigen expression between single individuals was noted. Blood group A type 2 and 3 (which is an extension of A type 2) antigens were present both as basic 6 and 9 sugar structures as well as extended saccharide chains migrating in the 8 to 11 sugar interval. In contrast, the type 1 chain based A and Lewis antigens were only present as their basic 5 to 7 sugar chains, and no elongated structures were found. Four cases of A2 kidneys initially transplanted into O recipients and removed after 5, 12, 21 days and 4 years, respectively, were also analyzed. Two of these kidneys, originating from the same donor, showed a difference in A antigen expression. The kidney functioning for four years (lost due to chronic rejection) completely lacked X antigen with five sugar residues (present in all other individuals) and contained a large amount of A antigens.(ABSTRACT TRUNCATED AT 250 WORDS)
ABO Blood-Group System↗
Plasma and red-cell glycolipid patterns of Le(a+b+) and Le(a+b-) Polynesians as further evidence of the weak secretor gene Se(w).
Monoclonal antibodies and thin-layer chromatography were used to study the unusual erythrocyte Lewis phenotypes found in healthy Polynesians. A single monoclonal anti-Leb reagent 073 (clone LM129) was found which could detect Leb antigen on the Polynesian erythrocytes of samples that were unreactive with various polyclonal and monoclonal anti-Leb reagents. Glycolipid fractions prepared from the plasma and erythrocytes of selected Polynesian samples of red-cell Le(a-b-), Le(a+b-) and Le(a+b+) phenotypes were found to have Leb glycolipids. The Leb antigen in some individuals is so weakly expressed that it is undetectable by routine erythrocyte phenotyping. Unusually large glycolipids bearing the Leb epitope were also found in some Polynesian samples, although the contribution of these novel glycolipids to phenotyping is unclear. The inability to detect Leb by routine methods and the presence of novel structures can be partially explained in terms of the presence of a weak secretor gene Se(w).
Chromatography, Thin Layer↗
The blood group B type-4 heptaglycosylceramide is a minor blood group B structure in human B kidneys in contrast to the corresponding A type-4 compound in A kidneys. Structural and in vitro biosynthetic studies.
Blood group A glycolipid antigens have been found based upon at least four different core saccharides (types 1 to 4). The biological significance of this structural polymorphism is not known, although the successful outcome of transplantations of blood group A2 kidneys to blood group O individuals have been partly explained by the low expression of A type-3 and -4 chain glycolipid antigens in A2 kidneys. If graft rejection due to ABO incompatibility is, in any way, correlated to the expression of type-3 and -4 chain blood group glycolipids, it is of interest to identify possible blood group B structures based on these core saccharides. In a non-acid glycosphingolipid fraction isolated from human blood group B kidneys, mass spectrometry, high-temperature gas chromatography-mass spectrometry and probing of thin-layer chromatograms with Gal alpha 1-4Gal-specific Escherichia coli and monoclonal anti-B antibodies provided evidence for minute amounts of a Gal alpha 1-3(Fuc alpha 1-2)Gal beta-HexNAc-Gal alpha 1-4Gal beta-Hex-Ceramide structure consistent with a B type-4 chain heptaglycosylceramide. In contrast, blood group A kidneys have the corresponding A type-4 chain heptaglycosylceramide as the predominant blood group A glycolipid. No, or very low activity of the blood group B gene enzyme on the type-4 chain blood group H hexaglycosylceramide precursor was found by biosynthetic experiments in vitro, which might explain the low expression of type-4 chain blood group B heptaglycosylceramides in human blood group B kidneys.
ABO Blood-Group System↗
Glycosphingolipid expression in spontaneously aborted fetuses and placenta from blood group p women. Evidence for placenta being the primary target for anti-Tja-antibodies.
A 12-week-old fetus and one 17-week-old fetus + placenta were obtained after spontaneous abortions from two women of blood group p. The 17-week-old fetus was dissected into intestine, liver, brain and residual tissue. Nonacid glycosphingolipid fractions were prepared from the tissues. Glycolipid characterization was carried out using thin layer chromatography immunostained with monoclonal antibodies and bacteria and by 1H NMR spectroscopy and mass spectrometry. In the placental fraction substantial amounts of globotetraosylceramide (P-antigen) and globotriaosylceramide (Pk-antigen) were identified. In contrast, the fetuses contained only trace amounts of these structures, as revealed by immunostaining. These results indicate that the primary target for the antibodies of the anti-Tja serum is the placenta tissue, resulting in termination of the pregnancy.
Abortion, Spontaneous↗
Characterisation of the anti-A antibody response following an ABO incompatible (A2 to O) kidney transplantation.
Anti-A,B antibodies produced in a blood group OLe(a-b-) recipient receiving a kidney graft from a blood group A2Le(a-b+) donor have been analysed for their ability to bind to different glycosphingolipid antigens. Solid-phase RIA using pure glycosphingolipid antigens and a chromatogram binding assay using total nonacid glycosphingolipid fractions from erythrocytes of different human blood group phenotypes together with pure glycolipid antigens were used as assay systems. Serum antibodies were shown to bind equally well to A (types 1, 2, 3 and 4) and B (types 1 and 2) antigenic structures but no binding to H antigens (types 1, 2 and 4) was detected. After adsorption of serum antibodies on A1 Le(a-b+) erythrocytes there was a residual anti-A antibody activity which could not be adsorbed by synthetic A-trisaccharides coupled to crystalline silica (Synsorb-A). These residual antibodies, which are not present in a pretransplant serum sample, had a specificity for the A antigen with type 1 core saccharide chain and the binding epitope obviously included both the N-acetylgalactosamine and the N-acetylglucosamine. The fucose residue was apparently not obligate for binding. The conformation of the sugar units involved in the binding epitope was determined.
ABO Blood-Group System↗
Serological and immunochemical characterization of anti-PP1Pk (anti-Tja) antibodies in blood group little p individuals. Blood group A type 4 recognition due to internal binding.
Serum samples from 13 blood group little p individuals were tested by radioimmunoassay for their IgG antibody subclass distribution against the P, P1 and Pk antigens. There was no uniform subclass distribution pattern, although all but one had IgG3 antibodies against all the P system antigens tested. Studies were performed adsorbing anti-Tja serum sequentially to columns with synthetic carbohydrate antigenic determinants within the P system coupled to silica beads (SynsorbsR). The effect on agglutinin and indirect antiglobulin titers was determined after adsorption to SynsorbsR with different P-system antigens (P1, Pk, P). Adsorption to all the three SynsorbsR was needed to eliminate or strongly reduce antibody titers. The effect on IgM, IgG, IgA as well as IgG subclass antibody binding to P, P1 and Pk antigens was also determined by radioimmunoassay and chromatogram binding assay. Anti-PP1Pk antibodies from a little p woman with repeated abortions were shown to bind to glycosphingolipid antigens prepared from one of the aborted placentae using a chromatogram binding assay. This binding was eliminated by serum adsorption to SynsorbsR with P1, Pk and P carbohydrates. Anti-PP1Pk antibodies were also shown to bind to extended structures in the globoseries, i.e. globopentaosylceramide, globohexaosylceramide (globo-H) and globoheptaosylceramide (globo-A). This binding is most probably due to antibodies recognizing internal sequences in the carbohydrate chain. Attempts were made to visualize the binding epitope of the antibodies by computer molecular modelling.
Adult↗
Alterations of glycosphingolipid-based blood group antigen expression on erythrocytes and in plasma studied on consecutive samples after a blood group O to A bone marrow transplantation.
A blood group A1Le(a-b+) individual with chronic myeloid leukaemia had received a bone marrow graft from an HLA-identical OLe(a+b-) donor. Twelve months after bone marrow transplantation (BMT), the red blood cells of the patient became agglutinable with anti-A blood group reagents. To elucidate whether the blood group A antigen expression was of plasma or of bone marrow origin, total non-acid glycosphingolipid fractions were prepared from red blood cells and plasma collected 17 months after BMT, and from plasma collected 13, 15 and 19 weeks after BMT. The glycolipid fractions were analysed by thin-layer chromatography and immunostained with monoclonal A-antibodies, and permethylated and permethylated-reduced derivatives of selected plasma samples were analysed by mass spectrometry. The results strongly indicate the presence of host bone marrow-produced blood group A red blood cells. Furthermore, the presence of a blood group H active pentaglycosylceramide type 1 (H-5-1) (Table I), characteristic for an OLe(a-b-) secretor, was seen in plasma 3-4 weeks before clinical chronic graft versus host disease (GVHD). After treatment of chronic GVHD, this expression disappeared. The blood group ALeb (A-7-1) antigen produced by the recipient seems to be present and to increase with time in all plasma samples. This also seems to be the case for the Leb and A-6-1 antigens.
ABO Blood-Group System↗
Non-acid glycosphingolipid expression in plasma of an A1 Le(a-b+) secretor human individual: identification of an ALeb heptaglycosylceramide as major blood group component.
Total non-acid glycosphingolipids were isolated from plasma of an A1 Le(a-b+) secretor individual with Refsum's disease (phytanic acid storage disease). The glycolipids were separated into 11 fractions by open column chromatography and by HPLC. The fractions were analyzed by thin-layer chromatography and tested for different blood group A activities as well as blood group Le(a )and Leb activity. The fractions were structurally characterized by proton NMR spectroscopy and FAB mass spectrometry and in selected cases by EI mass spectrometry of the permethylated and permethylated-reduced derivatives. Degradation analysis was performed on partially permethylated or permethylated-reduced alditol acetates. The dominating blood group compound was found to be a blood group A active type 1 chain difucosylheptaglycosylceramide. Other blood group compounds were identified as a blood group A active type 1 chain monofucosylhexaglycosylceramide, a blood group Leb hexaglycosylceramide, a blood group H active type 1 chain pentaglycosylceramide, and a globotetraosylceramide (the P-antigen). The presence of a Le(a) glycosphingolipid and blood group A type 3/4 chain structures were also found by immunostaining. Glucosyl-, lactosyl-, and globotriaosylceramides were the dominating short chain compounds. The amount of phytanic acid incorporated into the monoglycosylceramide fraction was found to be less than 5% of the fatty acids.
Animals↗
Basic biochemistry of cell surface carbohydrates and aspects of the tissue distribution of histo-blood group ABH and related glycosphingolipids.
Cell surface carbohydrates may be protein- or lipid-linked. The structural polymorphism of the oligosaccharide chains is extensive due to variations in monosaccharide composition, carbohydrate sequence, branching, linkage position and linkage anomericity. Blood group ABH and related glycosphingolipids show a remarkable tissue-specific expression with possible implications in areas such as transfusion medicine, transplantation surgery and oncology. This communication gives a condensed description of the present knowledge of the tissue-specific distribution of histo-blood glycolipids in humans.
ABO Blood-Group System↗
Carbohydrate specificity of human immunoglobulin-M antibodies with pig lymphocytotoxic activity.
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ABO Blood-Group System↗
Blood group type glycosphingolipids of human kidneys. Structural characterization of extended globo-series compounds.
Blood group type glycosphingolipids present in kidneys of blood group A and B human individuals have been isolated and structurally characterized by mass spectrometry, proton NMR spectroscopy, degradation studies and by their reactivity with various monoclonal antibodies and Escherichia coli bacteria. The two major complex glycolipids present in the blood group A and B kidneys were globopentaosylceramide (IV3Gal beta-Gb4Cer) and the X pentaglycosylceramide (III3Fuc alpha-nLc4Cer). The major blood group A glycolipid in the blood group A kidneys was based on the type 4 chain (globo-series). There were also small amounts of the type 2 chain and trace amounts of the type 1 and type 3 chain based A glycolipids. In addition, the blood group H type 4 chain structure was present together with Le(a) and Le(b) compounds. In the blood group B kidneys, the major B glycolipids were monofucosylated hexa- and octaglycosylceramides, where the former were based on the type 2 carbohydrate chain. The blood group B type 4 chain heptaglycosylceramide was found to be a minor component making up only about 1% of the total blood group B structures.
ABO Blood-Group System↗
Presence of glycosyltransferase inhibitors in the sera of patients with long-term surviving ABO incompatible (A2 to O) kidney grafts.
In the sera of blood group O recipients with long-term surviving blood group A2 kidney grafts, inhibitors of A-glycosyltransferase (alpha-3-N-acetyl-D-galactosaminyl transferase) have been identified in four out of six patients. The inhibitors seem to be of IgG antibody nature and do not appear until at least 1 year after transplantation. The inhibitors are also active against B-transferase (alpha-3-D-galactosyl transferase) as well as against A-glycosyltransferase of pig origin. The importance of these A-glycosyltransferase inhibitors is not clear. They may have a modifying effect on the antigenic expression of the graft. The immunogenicity of soluble glycosyltransferases is well known and the development of antibodies against them might prevent their enzymatic activity.
ABO Blood-Group System↗
Blood group A glycolipid antigen expression in kidney, ureter, kidney artery, and kidney vein from a blood group A1Le(a-b+) human individual. Evidence for a novel blood group A heptaglycosylceramide based on a type 3 carbohydrate chain.
Kidney, ureter, kidney artery, and kidney vein tissue were obtained from a single human transplant specimen. The donors erythrocyte blood group phenotype was A1Le(a-b+). Total non-acid glycolipid fractions were isolated and individual glycolipid components were identified by immunostaining thin layer plates with a panel of monoclonal antibodies and by mass spectrometry of the permethylated and permethylated-reduced total glycolipid fractions. The dominating glycolipids in all tissues were mono- to tetraglycosylceramides. In the kidney, ureter, and artery tissue less than 1% of the glycolipids were of blood group type, having more than 4 sugar residues. In contrast, 14% of the vein glycolipids were of blood group type, and the dominating components were type 1 chain blood group H pentaglycosylceramides and A hexaglycosylceramides. Trace amounts of structurally different blood group A glycolipids (type 1 to 4 core saccharide chains) with up to 10 sugar residues were found in the kidney, ureter, and vein tissues, including evidence for a novel blood group A heptaglycosylceramide based on the type 3 chain in the vein. The only detected A glycolipid antigen in the artery tissue was the blood group A difucosyl type 1 chain heptaglycosylceramide (ALeb) structure. Blood group Lewis and related antigens (Lea, Leb, and ALeb) were expressed in the kidney, ureter, and artery, but were completely lacking in the vein, indicating that the Le gene-coded alpha 1-4-fucosyltransferase was not expressed in this tissue. The X and Y antigens (type 2 chain isomers of the Lea and Leb antigens) were detected only in the kidney tissue.
ABO Blood-Group System↗
Glycolipid- and glycoprotein-based blood group A antigen expression in human thrombocytes. A1/A2 difference.
Total non-acid glycolipid fractions and total sodium dodecylsulphate (SDS) solubilized protein fractions were isolated from human thrombocytes obtained from single human donors having different blood group A1/A2 phenotypes. The blood group A glycolipid antigens were characterized by immunostaining of thin layer plates with different monoclonal anti-A antibodies. The glycoproteins carrying blood group A epitopes were identified by SDS-PAGE and Western blot analysis using a monoclonal anti-A antibody. Blood group A glycolipid antigens were found in both A1 and A2 thrombocytes but the A2 individuals expressed at least ten times less A glycolipids compared to the A1 individuals. Expression of A type 3/4 chain and small amounts of A type 1 chain glycolipids were seen in thrombocytes of both A1 and A2 individuals, while the type 2 chain A glycolipids appeared to be missing from the A2 thrombocytes. Blood group A reactive glycoproteins were only found in thrombocytes of A1 individuals and could not be detected in A2 individuals or a blood group O individual. The major blood group A glycoprotein were found as a double band migrating in the 130 kDa region.
ABO Blood-Group System↗
Blood group A glycolipid antigen biosynthesis: discrimination between biosynthesized and enzyme preparation derived blood group A antigen by mass spectrometry.
A monofucosyl type 1 chain blood group A hexaglycosylceramide was biosynthesized in solution using the type 1 chain blood group H pentaglycosylceramide as precursor, a crude microsomal fraction prepared from the mucosa scraping of a blood group A pig small intestine as enzyme source, and uridine diphosphate-N-acetyl-(1-14C)galactosamine as sugar donor. The radioactive product was enriched using reversed-phase column chromatography and silica gel HPLC. The peak, as detected by a beta-flow scintillation counter, was collected, permethylated, and analyzed by mass spectrometry. Carbohydrate sequence ions were found, indicating the presence of both the biosynthesized and a native, non-14C-containing blood group A hexaglycosylceramide. The blood group A pig small intestinal mucosa used as the enzyme source contain blood group A hexaglycosylceramide as the predominant glycolipid. Therefore, it is concluded that the nonradioactive blood group A hexaglycosylceramide found after the biosynthesis is derived from the enzyme preparation.
ABO Blood-Group System↗
Mass spectrometry of mixtures of intact glycosphingolipids.
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Carbohydrate Sequence↗