Cytokine-inducing macromolecular glycolipids from Enterococcus hirae: improved method for separation and analysis of its effects on cellular activation
| Autor: | Kiyoshi Takeda, Kazue Aoyama, T Tamura, Shoichi Kusumoto, Osamu Takeuchi, Toshiyuki Morichika, Masahito Hashimoto, Yasuo Suda, Shozo Kotani, Yoshimasa Imamura, Shizuo Akira, Kazumi Arimoto |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Lipopolysaccharides
medicine.medical_treatment Biophysics Lipopolysaccharide Receptors Receptors Cell Surface Biology Biochemistry Mice Glycolipid Calcitriol Enterococcus hirae medicine Tumor Cells Cultured Animals Drosophila Proteins Humans Receptor Molecular Biology Cells Cultured Mice Knockout Membrane Glycoproteins Interleukin-6 Tumor Necrosis Factor-alpha Macrophages Toll-Like Receptors Interleukin-18 Antibodies Monoclonal Cell Differentiation Cell Biology biology.organism_classification Toll-Like Receptor 2 Up-Regulation carbohydrates (lipids) Toll-Like Receptor 4 TLR2 Cytokine Monocytic leukemia Cytokines lipids (amino acids peptides and proteins) Tumor necrosis factor alpha Lipoteichoic acid Glycolipids Enterococcus |
| Zdroj: | Biochemical and biophysical research communications. 273(1) |
| ISSN: | 0006-291X |
| Popis: | Previously, we showed that several minor macromolecular glycolipids accounting for less than 5% of the lipoteichoic acid (LTA) fraction from Enterococcus hirae ATCC 9790 possess cytokine-inducing activity, whereas the purified LTA does not. In other words, the immunobiological activity of the LTA fraction reported in the 1980s was not attributable to LTA itself, but to other glycolipids coexisting in the fraction. In the present study, we improved the procedure of separation of the active glycolipids and evaluated their effects on cellular activation. The immunobiologically active glycolipids were separated from the crude glycolipid fraction obtained by hot phenol-water extraction of the cells. The total yield of active glycolipids was about fivefold higher than that separated by the previous method. Interleukin-6-inducing activities of the active glycolipids from 1,25-dihydroxy vitamin D 3 -differentiated human monocytic leukemia cells, THP-1, were inhibited by anti-CD14 mAbs in a dose-dependent manner. Macrophages from Toll-like receptor (TLR)-2-deficient or -4-deficient mice completely lacked the ability to produce tumor necrosis factor-α on stimulation with active glycolipids. These observations indicated that the cellular activation by the active glycolipids from E. hirae is mediated by CD14 and by both TLR2 and TLR4. |
| Databáze: | OpenAIRE |
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