| 1. |
- Barone, Angela, et al.
(författare)
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Characterization of acid and non-acid glycosphingolipids of porcine heart valve cusps as potential immune targets in biological heart valve grafts.
- 2014
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Ingår i: Xenotransplantation. - : Wiley. - 1399-3089 .- 0908-665X. ; 21:6, s. 510-522
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Tidskriftsartikel (refereegranskat)abstract
- Although xenotransplantation of vascularized organs/cells has not yet reached the clinic, glutaraldehyde-treated bioprosthetic heart valves (BHV), derived from porcine or bovine tissues, are today used for clinical replacement of diseased heart valves. However, the durability of these valve cusps is limited partly due to the onset of immune responses to the grafts. The xenoantigen-determinant Galα3Gal- and corresponding anti-Gal antibodies have been postulated to in part contribute to BHV damage. However, the presence of other non-Gal carbohydrate antigen determinants as well as the immune response to these non-Gal antigens and the inflammatory response generated by their interaction with the immune system has not been studied. In this study, we have isolated and structurally characterized both non-acid and acid glycosphingolipids from naïve porcine aortic and pulmonary valve cusps.
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| 2. |
- Barone, Angela, et al.
(författare)
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Evaluation of Sialyl-Lactotetra as a Marker for Epithelial Ovarian Tumors
- 2020
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Ingår i: Frontiers in Oncology. - : Frontiers Media SA. - 2234-943X. ; 10
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Tidskriftsartikel (refereegranskat)abstract
- Ovarian carcinoma is a heterogeneous disease with distinct molecular and histological profiles, ranging from low grade atypia to highly aggressive tumors associated with a poor prognosis. In the present study, glycosphingolipids were isolated from human high-grade serous ovarian carcinoma, whereby the novel stem cell marker Sialyl-lactotetra (S-Lc(4)) was characterized in two out of three cases. The presence and level of S-Lc(4)was further evaluated immunohistochemically in a cohort of patients with ovarian tumors ranging from benign lesions to high grade serous carcinoma (n= 478). Its expression was assessed in association with tumor grade, stage, histology, and survival. The data showed that S-Lc(4)is most common and highly expressed in borderline type tumors and carcinomas with low levels of aggressiveness, such as mucinous, endometrioid, and low grade serous. Accordingly, S-Lc(4)-positivity was associated with better disease-free survival. The expression of S-Lc(4)was seemingly associated with lineage continuity and could be traced from premalignant lesions to carcinoma, suggesting inheritance by a stem cell lineage that gives rise to generally indolent tumors.
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| 3. |
- Barone, Angela, et al.
(författare)
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Glycosphingolipids of porcine, bovine, and equine pericardia as potential immune targets in bioprosthetic heart valve grafts
- 2018
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Ingår i: Xenotransplantation. - : Wiley. - 0908-665X. ; 25:5
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Tidskriftsartikel (refereegranskat)abstract
- BackgroundPericardial tissue from various animal species is utilized for the production of the bioprosthetic heart valves (BHV) used clinically. Experimental data show that the eventual breakdown of BHV is partly due to immunological interactions with carbohydrate tissue antigens. To understand these processes, we have examined the glycolipid-based carbohydrate antigens in naive porcine, bovine, and equine pericardia. ExperimentalTotal non-acid and acid glycosphingolipid fractions were isolated from porcine, bovine, and equine pericardia, and individual glycolipid compounds were characterized by thin-layer chromatography, mass spectrometry, and binding of monoclonal antibodies, lectins and bacteria in chromatogram binding assays. ResultsThe non-acid glycolipid fractions from all species contained glycosphingolipids based on the globo- and neolacto-series, including pentaglycosylceramides with terminal Gal3 determinants. Terminal blood group A and H (O) structures based on type 2 core chains were present in porcine pericardium, while the Forssman pentaosylceramide was found in equine pericardium. All acid glycolipid fractions contained sulfatide and several gangliosides with both N-acetyl- and N-glycolyl-neuraminic acid as terminal saccharide chain determinants. ConclusionSeveral carbohydrate antigens which are potential targets for the human immune system have been identified in the animal pericardial tissues used for the production of BHV. Which of these antigens are left in the tissues after industrial BHV production processes, as well as their potential role in eventual BHV degradation, remains to be elucidated.
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| 4. |
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| 5. |
- Barone, Angela, et al.
(författare)
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Structural Complexity of Non-acid Glycosphingolipids in Human Embryonic Stem Cells Grown under Feeder-free Conditions.
- 2013
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Ingår i: The Journal of biological chemistry. - 1083-351X. ; 288:14, s. 10035-50
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Tidskriftsartikel (refereegranskat)abstract
- Due to their pluripotency and growth capability, there are great expectations for human embryonic stem cells, both as a resource for functional studies of early human development and as a renewable source of cells for use in regenerative medicine and transplantation. However, to bring human embryonic stem cells into clinical applications, their cell surface antigen expression and its chemical structural complexity have to be defined. In the present study, total non-acid glycosphingolipid fractions were isolated from two human embryonic stem cell lines (SA121 and SA181) originating from leftover in vitro fertilized human embryos, using large amounts of starting material (1 × 10(9) cells/cell line). The total non-acid glycosphingolipid fractions were characterized by antibody and lectin binding, mass spectrometry, and proton NMR. In addition to the globo-series and type 1 core chain glycosphingolipids previously described in human embryonic stem cells, a number of type 2 core chain glycosphingolipids (neo-lactotetraosylceramide, the H type 2 pentaosylceramide, the Le(x) pentaosylceramide, and the Le(y) hexaosylceramide) were identified as well as the blood group A type 1 hexaosylceramide. Finally, the mono-, di-, and triglycosylceramides were characterized as galactosylceramide, glucosylceramide, lactosylceramide, galabiaosylceramide, globotriaosylceramide, and lactotriaosylceramide. Thus, the glycan diversity of human embryonic stem cells, including cell surface immune determinants, is more complex than previously appreciated.
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| 6. |
- Barone, Angela
(författare)
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Studies on glycosphingolipids in regenerative medicine
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Regenerative medicine, including stem cell based therapies and xenotransplantation, is a new and developing field that aims to restore normal function in end stage cell/organ failure. However, in therapeutic settings involving transfer of cells with a different genetic background, these cells will expose the recipient to non-self cell surface antigenic determinants that may evoke an immune response and subsequent graft damage. Thus, the cell surface antigen expression of cells aimed for transplantation has to be defined. The first part of this study deals with glycosphingolipids of human embryonic stem cells (hESC). The glycosphingolipids were isolated from a relatively high number of cells (1x109 cells/cell line) allowing separation of the glycosphingolipids into total non-acid and acid fractions, that could be further separated into sub-fractions. These fractions were structurally characterized by mass spectrometry, proton NMR spectroscopy and binding studies with carbohydrate binding ligands. This allowed identification of several glycosphingolipids not previously described in hESC. In the non-acid glycosphingolipid fractions several novel blood group H, Lex and Ley compounds based on neolacto core chains were characterized, in addition to the already identified lacto- and globo-series glycosphingolipids. The acidic glycosphingolipid fractions contained several novel hESC acid glycosphingolipids, like the gangliosides sialyl-lactotetraosylceramide and sialyl-globotetraosylceramide, and the sulfated glycosphingolipids sulfatide, sulf-lactosylceramide and sulf-globopentaosylceramide. The cellular and subcellular distribution of sialyl-lactotetraosylceramide and sulfated glycosphingolipids in hESC and in human induced pluripotent stem cells (hiPSC) was explored by flow cytometry, immunohistochemistry and electron microscopy. A high cell surface expression of sialyl-lactotetra on hESC and hiPSC was demonstrated, whereas the sulfated glycosphingolipids were restricted to intracellular compartments. During differentiation of hiPSC into hepatocyte-like cells a rapid down-regulation of the sialyl-lactotetra epitope was found. Taken together these data demonstrate that the sialyl-lactotetra carbohydrate sequence is a novel marker for undifferentiated human pluripotent stem cells. Diseased human heart valves are substituted with either mechanical valves or biological heart valves (BHV) produced from porcine and bovine valves or pericardial tissues. The BHV function deteriorates with time partly due to an immunological process. The second part of the study aimed at defining carbohydrate antigens of porcine heart valves with a potential of being immune targets for this process. Here, a number of acidic glycosphingolipids (sulfatide and the gangliosides GM3, GM2, GM1, fucosyl-GM1, GD3 and GD1a) and non-acid glycosphingolipids (globotetraosylceramide, H type 2 pentaosylceramide, fucosyl-gangliotetraosylceramide and Galα3neolacto-tetraosylceramide) were characterized. Interestingly, no gangliosides with the non-Gal xenoantigen NeuGc were found. However, the Galα3 epitope is the major xenoantigen, and thus a possible target for antibody mediated immune reactions to xenogeneic bioprosthetic heart valves in humans.
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| 7. |
- Benktander, John, et al.
(författare)
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Helicobacter pylori SabA binding gangliosides of human stomach
- 2018
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Ingår i: Virulence. - : Informa UK Limited. - 2150-5594 .- 2150-5608. ; 9:1, s. 738-751
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Tidskriftsartikel (refereegranskat)abstract
- Adhesion of Helicobacter pylori to the gastric mucosa is a prerequisite for the pathogenesis of H. pylori related diseases. In this study, we investigated the ganglioside composition of human stomach as the target for attachment mediated by H. pylori SabA (sialic acid binding adhesin). Acid glycosphingolipids were isolated from human stomach and separated into subfractions, which were characterized by mass spectrometry and by binding of antibodies, bacteria, and Solanum tuberosum lectin. H. pylori SabA binding gangliosides were characterized as Neu5Ac alpha 3-neolactohexaosylceramide and Neu5Ac alpha 3-neolactooctaosylceramide, while the other acid human stomach glycosphingolipids characterized (sulfatide and the gangliosides GM3, GD3, GM1, Neu5Ac alpha 3-neolactotetraosylceramide, GD1a and GD1b) were not recognized by the bacteria. Defining H. pylori binding glycosphingolipids of the human gastric mucosa will be useful to specifically target this microbe-host interaction for therapeutic intervention.
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| 8. |
- Breimer, Michael, 1951, et al.
(författare)
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Glycosphingolipids of human embryonic stem cells.
- 2017
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Ingår i: Glycoconjugate journal. - : Springer Science and Business Media LLC. - 1573-4986 .- 0282-0080. ; 34:6, s. 713-723
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Forskningsöversikt (refereegranskat)abstract
- The application of human stem cell technology offers theoretically a great potential to treat various human diseases. However, to achieve this goal a large number of scientific issues remain to be solved. Cell surface carbohydrate antigens are involved in a number of biomedical phenomena that are important in clinical applications of stem cells, such as cell differentiation and immune reactivity. Due to their cell surface localization, carbohydrate epitopes are ideally suited for characterization of human pluripotent stem cells. Amongst the most commonly used markers to identify human pluripotent stem cells are the globo-series glycosphingolipids SSEA-3 and SSEA-4. However, our knowledge regarding human pluripotent stem cell glycosphingolipid expression was until recently mainly based on immunological assays of intact cells due to the very limited amounts of cell material available. In recent years the knowledge regarding glycosphingolipids in human embryonic stem cells has been extended by biochemical studies, which is the focus of this review. In addition, the distribution of the human pluripotent stem cell glycosphingolipids in human tissues, and glycosphingolipid changes during human stem cell differentiation, are discussed.
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| 9. |
- Esberg, Anders, et al.
(författare)
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Corynebacterium matruchotii Demography and Adhesion Determinants in the Oral Cavity of Healthy Individuals
- 2020
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Ingår i: Microorganisms. - : MDPI. - 2076-2607. ; 8:11
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Tidskriftsartikel (refereegranskat)abstract
- Corynebacterium matruchotii may be key in tooth biofilm formation, but information about demographics, bacterial partners, and binding ligands is limited. The aims of this study were to explore C. matruchotii's demography by age and colonization site (plaque and saliva), in vitro bacterial-bacterial interactions in coaggregation and coadhesion assays, and glycolipids as potential binding ligands in thin-layer chromatogram binding assays. C. matruchotii prevalence increased from 3 months to 18 years old, with 90% and 100% prevalence in saliva and tooth biofilm, respectively. C. matruchotii aggregated in saliva in a dose-dependent manner but lacked the ability to bind to saliva-coated hydroxyapatite. In vivo, C. matruchotii abundance paralleled that of Actinomyces naeslundii, Capnocytophaga sp. HMT 326, Fusobacterium nucleatum subsp. polymorphum, and Tannerella sp. HMT 286. In vitro, C. matruchotii bound both planktonic and surface-bound A. naeslundii, Actinomyces odontolyticus, and F. nucleatum. In addition, C. matruchotii exhibited the ability to bind glycolipids isolated from human erythrocytes (blood group O), human granulocytes, rabbit intestine, human meconium, and rat intestine. Binding assays identified candidate carbohydrate ligands as isoglobotriaosylceramide, Gal alpha 3-isoglobotriaosylceramide, lactotriaosylceramide, lactotetraosylceramide, neolactotetraosylceramide, and neolactohexaosylceramide. Thus, C. matruchotii likely uses specific plaque bacteria to adhere to the biofilm and may interact with human tissues through carbohydrate interactions.
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| 10. |
- Jin, Chunsheng, et al.
(författare)
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Helicobacter pylori-binding nonacid glycosphingolipids in the human stomach
- 2018
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Ingår i: Journal of Biological Chemistry. - : American Society for Biochemistry and Molecular Biology. - 0021-9258 .- 1083-351X. ; 293:44, s. 17248-17266
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Tidskriftsartikel (refereegranskat)abstract
- Helicobacter pylori has a number of well-characterized carbohydrate-binding adhesins (BabA, SabA, and LabA) that promote adhesion to the gastric mucosa. In contrast, information on the glycoconjugates present in the human stomach remains unavailable. Here, we used MS and binding of carbohydrate-recognizing ligands to characterize the glycosphingolipids of three human stomachs from individuals with different blood group phenotypes (O(Rh-)P, A(Rh+)P, and A(Rh+)p), focusing on compounds recognized by H. pylori. We observed a high degree of structural complexity, and the composition of glycosphingolipids differed among individuals with different blood groups. The type 2 chain was the dominating core chain of the complex glycosphingolipids in the human stomach, in contrast to the complex glycosphingolipids in the human small intestine, which have mainly a type 1 core. H. pylori did not bind to the O(Rh-)P stomach glycosphingolipids, whose major complex glycosphingolipids were neolactotetraosylceramide, the Le(x), Le(a), and H type 2 pentaosylceramides, and the Le(y) hexaosylceramide. Several H. pylori-binding compounds were present among the A(Rh+)P and A(Rh+)p stomach glycosphingolipids. Ligands for BabA-mediated binding of H. pylori were the Le(b) hexaosylceramide, the H type 1 pentaosylceramide, and the A type 1/ALe(b) heptaosylceramide. Additional H. pylori-binding glycosphingolipids recognized by BabA-deficient strains were lactosylceramide, lactotetraosylceramide, the x(2) pentaosylceramide, and neolactohexaosylceramide. Our characterization of human gastric receptors required for H. pylori adhesion provides a basis for the development of specific compounds that inhibit the binding of this bacterium to the human gastric mucosa.
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