Macrophage Recognition of Thiol-Group Oxidized Cells: Recognition of Carbohydrate Chains by Macrophage Surface Nucleolin as Apoptotic Cells
Autor: | Yuichi Miki, Kazuya Hirano, Masatoshi Beppu |
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Rok vydání: | 2012 |
Předmět: |
Apoptosis
Ligands Applied Microbiology and Biotechnology Biochemistry Jurkat cells Analytical Chemistry Jurkat Cells Western blot medicine Humans Macrophage Sulfhydryl Compounds Molecular Biology Polyacrylamide gel electrophoresis Diamide biology medicine.diagnostic_test Macrophages Organic Chemistry RNA-Binding Proteins hemic and immune systems General Medicine Phosphoproteins Molecular biology biology.protein Carbohydrate Metabolism DNA fragmentation Antibody Oxidation-Reduction Nucleolin Protein Binding Biotechnology |
Zdroj: | Bioscience, Biotechnology, and Biochemistry. 76:2068-2074 |
ISSN: | 1347-6947 0916-8451 |
Popis: | The mechanism was investigated for macrophage recognition of cells oxidized by diamide, a thiol group-specific oxidizing reagent. Jurkat cells exposed to various concentrations of diamide were recognized by macrophages, the cells exposed to 25 µM diamide being best recognized. CD43, a major glycoprotein on the Jurkat cell surface, tended to form clusters upon diamide oxidization, and pretreating Jurkat cells with the anti-CD43 antibody inhibited macrophage binding. This indicates that macrophages appeared to recognize CD43. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and a Western blot analysis of CD43 of the diamide-oxidized cells showed no increase in the amount of cross-linked CD43 compared with control cells, indicating that cross-linking of CD43 by a disulphide bond was not involved in the clustering. Both CD43 clustering and binding of the oxidized cells to macrophages was prevented by the caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethyl ketone (Z-VAD-fmk), suggesting that the oxidized and macrophage-bound cells were undergoing apoptosis. A closer examination revealed that the caspase-3 activity, chromatin condensation, and DNA fragmentation in Jurkat cells were all increased by oxidation. The macrophage receptor involved in the binding appeared to be the cell-surface protein, nucleolin; an anti-nucleolin antibody treatment inhibited the binding. These results suggest that thiol group-oxidized cells underwent early apoptosis and were recognized by nucleolin on macrophages as early apoptotic cells. |
Databáze: | OpenAIRE |
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