Energy dispersive spectroscopy‐scanning transmission electron microscope observations of free radical production in human polymorphonuclear leukocytes phagocytosing non‐opsonized <i>Tannerella forsythia</i>
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- Keiichi Moriguchi
- Department of Oral Anatomy, School of Dentistry Aichi‐Gakuin University Nagoya Aichi 464‐8650 Japan
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- Yoshiaki Hasegawa
- Department of Microbiology, School of Dentistry Aichi‐Gakuin University Nagoya Aichi 464‐8650 Japan
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- Naoya Higuchi
- Department of Endodontics, School of Dentistry Aichi‐Gakuin University Nagoya Aichi 464‐8650 Japan
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- Yukitaka Murakami
- Department of Oral Microbiology, Division of Oral Infections and Health Science Asahi University School of Dentistry 1851 Hozumi Mizuho Gifu 501‐0296 Japan
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- Fuminobu Yoshimura
- Department of Microbiology, School of Dentistry Aichi‐Gakuin University Nagoya Aichi 464‐8650 Japan
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- Kazuhiko Nakata
- Department of Endodontics, School of Dentistry Aichi‐Gakuin University Nagoya Aichi 464‐8650 Japan
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- Masaki Honda
- Department of Oral Anatomy, School of Dentistry Aichi‐Gakuin University Nagoya Aichi 464‐8650 Japan
書誌事項
- 公開日
- 2017-04-24
- 資源種別
- journal article
- 権利情報
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- http://onlinelibrary.wiley.com/termsAndConditions#vor
- DOI
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- 10.1002/jemt.22819
- 公開者
- Wiley
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説明
<jats:title>Abstract</jats:title><jats:p>We investigated the association between human polymorphonuclear leukocytes (PMNs) and non‐opsonized <jats:italic>Tannerella forsythia</jats:italic> ATCC 43037 displaying a serum‐resistant surface layer (S‐layer). When PMNs were mixed with <jats:italic>T. forsythia</jats:italic> in suspension, the cells phagocytosed <jats:italic>T. forsythia</jats:italic> cells. Nitro blue tetrazolium (NBT) reduction, indicative of <jats:inline-graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="graphic/jemt22819-math-0001.png" xlink:title="urn:x-wiley:1059910X:media:jemt22819:jemt22819-math-0001"/> production, was observed by light microscopy; cerium (Ce) perhydroxide deposition, indicative of H<jats:sub>2</jats:sub>O<jats:sub>2</jats:sub> production, was observed by electron microscopy. We examined the relationship between high‐molecular‐weight proteins of the S‐layer and Ce reaction (for <jats:italic>T. forsythia</jats:italic> phagocytosis) using electron microscopic immunolabeling. Immunogold particles were localized within the PMNs and on cell surfaces, labelling at the same Ce‐reacted sites where the S‐layer was present. We then used energy dispersive spectroscopy (EDS)‐scanning transmission electron microscope (STEM) to perform Ce and nitrogen (N) (for S‐layer immunocytochemistry) elemental analysis on the phagocytosed cells. That is, the elemental mapping and analysis of N by EDS appeared to reflect the presence of the same moieties detected by the 3,3′‐diaminobenzidine‐tetrahydrochloride (DAB) reaction with horseradish peroxidase (HRP)‐conjugated secondary antibodies, instead of immunogold labeling. We focused on the use of EDS‐STEM to visualize the presence of N resulting from the DAB reaction. In a parallel set of experiments, we used EDS‐STEM to perform Ce and gold (Au; from immunogold labeling of the S‐layer) elemental analysis on the same phagocytosing cells.</jats:p>
収録刊行物
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- Microscopy Research and Technique
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Microscopy Research and Technique 80 (6), 555-562, 2017-04-24
Wiley
