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Isolating the influences of fluid dynamics on selectin-mediated particle rolling at venular junctional regions.
Jung JJ, Grayson KA, King MR, Lamkin-Kennard KA
(2018) Microvasc Res 118: 144-154
MeSH Terms: Animals, Cell Adhesion, Humans, Hydrodynamics, Inflammation, Leukocyte Rolling, Leukocytes, Lewis X Antigen, Microspheres, Models, Cardiovascular, P-Selectin, Sialyl Lewis X Antigen, Signal Transduction, Venules
Show Abstract · Added April 15, 2019
The objective of this study was to isolate the impact of hydrodynamics on selectin-mediated cell rolling in branched microvessels. Significant advancements have been made in furthering the understanding of complex interactions between biochemical and physical factors in the inflammatory cascade in simplified planar geometries. However, few studies have sought to quantify the effects of branched configurations and to isolate the effects of associated fluid forces. Experimental techniques were developed to perform in vitro adhesion experiments in Y-shaped micro-slides. The micro-slides were coated with P-selectin and microspheres coated with Sialyl-Lewis were observed as they rolled in the chambers at different wall shear stresses. Study results revealed that microsphere rolling velocities and rolling flux were lowest in regions closest to the apex of a junctional region and were dependent on both branch angle and wall shear stress. The regions closest to the junctional region were shown to have low bulk flow velocities and shear stresses using computational fluid dynamics (CFD) modeling. Collectively, the study demonstrates that despite the presence of a uniform coating of P-selectin, hydrodynamic factors associated with the chamber geometry yield non-uniform effects on particle behavior. These findings could explain why cells have been observed to preferentially adhere or transmigrate near junctional regions. Future characterization of inflammatory processes in microvascular network configurations is therefore crucial for furthering our fundamental understanding of inflammation.
Copyright © 2018 Elsevier Inc. All rights reserved.
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14 MeSH Terms
Phenotypic diversity in Lewis expression of Helicobacter pylori isolates from the same host.
Wirth HP, Yang M, Peek RM, Höök-Nikanne J, Fried M, Blaser MJ
(1999) J Lab Clin Med 133: 488-500
MeSH Terms: Animals, Antigens, Bacterial, Bacterial Proteins, Female, Gene Expression Regulation, Bacterial, Genotype, Gerbillinae, Helicobacter Infections, Helicobacter pylori, Humans, Lewis Blood Group Antigens, Lewis X Antigen, Male, Mice, Phenotype, Polymerase Chain Reaction, Polymorphism, Genetic, Stomach
Show Abstract · Added March 5, 2014
Populations of Helicobacter pylori cells show a stable expression of Lewis surface antigens, although phase variation may occur among individual organisms grown in vitro. We searched for variation in Lewis phenotypes among H. pylori cells of minimally in vitro-passaged isolates. Lewis expression in 180 clonal H. pylori populations from the primary culture of 20 gastric biopsy samples from 12 patients, and that in 160 isolates from primary cultures from 16 experimentally infected rodents, were examined by enzyme immunoassays. Substantial differences in Lewis expression were found among the isolates from 9 (75%) of 12 patients. These differences were unrelated to overall genetic diversity as determined by polymerase chain reactions for random amplified polymorphic DNA or cagA status, and they persisted during subsequent in vitro passage. In contrast, Lewis expression was highly uniform in H. pylori isolates from different rodents infected for up to 20 weeks. Variation in H. pylori Lewis expression in genetically closely related organisms in human subjects may provide a pool of bacterial phenotypes for the continuous selection of optimally host-adapted populations suitable for persistence.
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18 MeSH Terms
Acid-induced expression of an LPS-associated gene in Helicobacter pylori.
McGowan CC, Necheva A, Thompson SA, Cover TL, Blaser MJ
(1998) Mol Microbiol 30: 19-31
MeSH Terms: Amino Acid Sequence, Bacterial Proteins, Electrophoresis, Polyacrylamide Gel, Gene Expression Regulation, Bacterial, Genes, Bacterial, Helicobacter pylori, Hydrogen-Ion Concentration, Immunoblotting, Lewis Blood Group Antigens, Lewis X Antigen, Lipopolysaccharides, Molecular Sequence Data, Nucleic Acid Hybridization, O Antigens, Sequence Analysis, DNA
Show Abstract · Added March 5, 2014
To investigate urease-independent mechanisms by which Helicobacter pylori resists acid stress, subtractive RNA hybridization was used to identify H. pylori genes whose expression is induced after exposure to acid pH. This approach led to the isolation of a gene that encoded a predicted 34.8kDa protein (WbcJ), which was homologous to known bacterial O-antigen biosynthesis proteins involved in the conversion of GDP-mannose to GDP-fucose. An isogenic wbcJ null mutant strain failed to express O-antigen and Lewis X or Lewis Y determinants and was more sensitive to acid stress than was the wild-type strain. Qualitative differences in LPS profiles were observed in H. pylori cells grown at pH 5 compared with pH 7, which suggests that H. pylori may alter its LPS structure in response to acidic pH. This may be an important adaptation facilitating H. pylori colonization of the acidic gastric environment.
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15 MeSH Terms
Ratio of IgG:IgM antibodies to sialyl Lewis(x) and GM3 correlates with tumor growth after immunization with melanoma-cell vaccine with different adjuvants in mice.
Ravindranath MH, Kelley MC, Jones RC, Amiri AA, Bauer PM, Morton DL
(1998) Int J Cancer 75: 117-24
MeSH Terms: Animals, Antibodies, Neoplasm, BCG Vaccine, Cancer Vaccines, Cell Division, Disease Progression, Enzyme-Linked Immunosorbent Assay, G(M3) Ganglioside, Humans, Immunoglobulin G, Immunoglobulin M, Lewis Blood Group Antigens, Lewis X Antigen, Male, Melanoma, Experimental, Mice, Mice, Inbred C57BL
Show Abstract · Added February 16, 2016
Human melanoma cells (from biopsies and culture) express sialyl-Lewis(x) and sialyl Lewis(a), the ligands for ECAM. These ligands may facilitate tumor progression and metastasis in human cancers. To test whether the antibodies to these ligands inhibit tumor progression, IgG and IgM responses to sLe(x) and sLe(a) were induced in C57BL/6j mice (n = 76) by immunization with human melanoma cells, with or without adjuvants (BCG, MPL, KLH). Control mice were treated with saline or BCG. Tumor growth and antigen expression were monitored after challenge with B16 mouse melanoma cells expressing sLe(x), sLe(a) and the ganglioside GM3. Tumor growth was reduced in mice immunized with BCG alone or cells with BCG or MPL, while tumors in mice receiving cells without adjuvants grew larger than in the control. Augmentation of IgM titers to sLe(x) and GM3 after immunization with BCG, or with cells with BCG or MPL correlated with retarded tumor growth, while increased IgG titers to sLe(x) significantly correlated with aggressive tumor growth in mice immunized with cells without adjuvants. SLe(x), sLe(a) and GM3 were expressed in tumors from mice treated with saline or BCG. SLe(x) expression, in particular, was lost in tumors growing in mice immunized with cells with or without adjuvants. Anti-sLe(x) antibodies may promote or prevent tumor growth by antigenic modulation or by cytotoxic killing of tumor cells. Since early anti-sLe(x) IgM correlated with tumor regression, in contrast to anti-sLe(x) IgG, it may serve as a potential early endpoint for the effectiveness of melanoma vaccines expressing the antigens.
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17 MeSH Terms
Helicobacter pylori Lewis expression is related to the host Lewis phenotype.
Wirth HP, Yang M, Peek RM, Tham KT, Blaser MJ
(1997) Gastroenterology 113: 1091-8
MeSH Terms: Adult, Aged, Aged, 80 and over, Biopsy, Erythrocytes, Female, Gastric Mucosa, Gastritis, Genotype, Helicobacter Infections, Helicobacter pylori, Humans, Inflammation, Lewis Blood Group Antigens, Lewis X Antigen, Male, Middle Aged, Phenotype, Pyloric Antrum
Show Abstract · Added March 5, 2014
BACKGROUND & AIMS - Lewis antigens occur in human gastric epithelium and in Helicobacter pylori lipopolysaccharide; their expression is polymorphic in both. Autoimmune mechanisms induced by bacterial Lewis expression have been proposed to cause gastritis. The aim of this study was to examine the relationship between bacterial and host gastric Lewis expression, as determined by the erythrocyte Lewis(a/b) phenotype, and between gastric histopathology and bacterial Lewis expression.
METHODS - H. pylori Lewis expression was determined by enzyme immunoassays, erythrocyte Lewis phenotype was assessed by agglutination tests, and gastric histopathology was scored blindly.
RESULTS - The host Lewis phenotype was (a+b-) in 15, (a-b+) in 34, and (a-b-) in 17 patients, therefore expressing Lewis x, y, or neither as their major gastric epithelial Lewis type 2 antigen. H. pylori from patients with Lewis(a+b-) expressed Lewis x more than y (1147 +/- 143 vs. 467 +/- 128 optical density units [ODU]; P = 0.006), isolates from patients with Lewis(a-b+) expressed Lewis x less than y (359 +/- 81 vs. 838 +/- 96 ODU; P = 0.0001), and isolates from Lewis(a-b-) patients expressed Lewis x and y approximately equally. Gastritis was unrelated to H. pylori Lewis expression.
CONCLUSIONS - In mimicking host gastric epithelium, H. pylori cells not only express Lewis x and y, but the relative proportion of expression corresponds to the host Lewis phenotype, suggesting selection for host-adapted organisms.
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19 MeSH Terms
Endothelial-selectin ligands sialyl Lewis(x) and sialyl Lewis(a) are differentiation antigens immunogenic in human melanoma.
Ravindranath MH, Amiri AA, Bauer PM, Kelley MC, Essner R, Morton DL
(1997) Cancer 79: 1686-97
MeSH Terms: Animals, Antibodies, Neoplasm, Antigens, Differentiation, Antigens, Neoplasm, Biopsy, CA-19-9 Antigen, Cancer Vaccines, Cell Transformation, Neoplastic, Endothelium, Vascular, Gangliosides, Humans, Ligands, Melanocytes, Melanoma, Mice, Oligosaccharides, Selectins, Sialyl Lewis X Antigen, Tumor Cells, Cultured
Show Abstract · Added February 16, 2016
BACKGROUND - Sialyl Lewis(x) (sLe(x)) and sialyl Lewis(a) (sLe(a)), the endothelial-selectin ligands involved in extravasation of neutrophils and carcinomas, have been identified in human melanoma. This study explored the following issue: If these ligands are immunogenic tumor-differentiation antigens, they would be potential targets for immunotherapy because of their putative roles in extravasation and metastasis.
METHODS - Using a cell-suspension enzyme-linked immunosorbent assay (ELISA), the expression of sLe(x) and sLe(a) on the surface of normal melanocytes, melanoma cells from biopsies, and cell lines (M10-v, M24, and M101) constituting melanoma cell vaccine (MCV) were quantitated. Melanoma patients were immunized with the MCV expressing these antigens. Sera of normal individuals, sera of patients, and sera that adsorbed to sLe(x) and sLe(a) were titrated for anti-sLe antibodies by ELISA to verify the immunogenicity of the ligands.
RESULTS - The normal melanocytes did not express sLe(x) and poorly expressed sLe(a). Melanoma cells from tumor biopsies and MCV lines expressed both sLe(x) and sLe(a). Sialyl Le(x) was associated with glycoprotein(s) in M10-v, and sLe(a) occurred as a glycolipid moiety in M24. MCV recipients developed high titers for immunoglobulin (Ig)M but not IgG to both ligands. IgM titers to these ligands were low in normal subjects. In some of the preimmune sera of patients, the titers were threefold above normal. Six of 13 MCV recipients developed at least a twofold increase in anti-sLe titers above preimmune level after the second or third immunization. Adsorption studies suggested that both ligands were immunogenic.
CONCLUSIONS - The melanoma-associated sLe(x) and sLe(a) are immunogenic neoplasm-differentiation antigens and are therefore potential targets for passive and active specific immunotherapy in the treatment of melanoma.
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19 MeSH Terms