Specific in situ visualization of plasma cells producing antibodies against porphyromonas gingivalis in gingival radicular cyst

Application of the enzyme-labeled antigen method

Shinya Tsuge, Yasuyoshi Mizutani, Kazuhiro Matsuoka, Tatsuya Sawasaki, Yaeta Endo, Koji Naruishi, Hiroshi Maeda, Shogo Takashiba, Kazuya Shiogama, Ken ichi Inada, Yutaka Tsutsumi

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The enzyme-labeled antigen method was applied to visualize plasma cells producing antibodies to Porphyromonas gingivalis, flora of the human oral cavity. Antibodies to P. gingivalis have reportedly been detected in sera of patients with periodontitis. Biotinylated bacterial antigens, Ag53, and four gingipain domains (Arg-pro, Arg-hgp, Lys-pro, and Lys-hgp) were prepared by the cell-free protein synthesis system using the wheat germ extract. In paraformaldehyde-fixed frozen sections of rat lymph nodes experimentally immunized with Ag53-positive and Ag53-negative P. gingivalis, plasma cells were labeled with biotinylated Arg-hgp and Lys-hgp. Antibodies to Ag53 were detected only in the nodes immunized with Ag53-positive bacteria. In two of eight lesions of gingival radicular cyst with inflammatory infiltration, CD138-positive plasma cells in frozen sections were signalized for Arg-hgp and Lys-hgp. An absorption study using unlabeled antigens confirmed the specificity of staining. The AlphaScreen method identified the same-type antibodies in tissue extracts but not in sera. Antibodies to Ag53, Arg-pro, and Lys-pro were undetectable. In two cases, serum antibodies to Arg-hgp and Lys-hgp were AlphaScreen positive, whereas plasma cells were scarcely observed within the lesions. These findings indicate the validity of the enzyme-labeled antigen method. This is the very first application of this novel histochemical technique to human clinical samples.

Original languageEnglish
Pages (from-to)673-689
Number of pages17
JournalJournal of Histochemistry and Cytochemistry
Volume59
Issue number7
DOIs
Publication statusPublished - 2011

Fingerprint

Radicular Cyst
Porphyromonas gingivalis
Plasma Cells
Antigens
arginylproline
lysylproline
Antibodies
Enzymes
Frozen Sections
Serum
Bacterial Antigens
Tissue Extracts
Periodontitis
Triticum
Mouth
Lymph Nodes
Staining and Labeling
Bacteria

Keywords

  • Ag53
  • Alphascreen method
  • Cell-free protein synthesis
  • Enzyme-labeled antigen method
  • Gingipains
  • Porphyromonas gingivalis
  • Radicular cyst

ASJC Scopus subject areas

  • Anatomy
  • Histology
  • Medicine(all)

Cite this

Specific in situ visualization of plasma cells producing antibodies against porphyromonas gingivalis in gingival radicular cyst : Application of the enzyme-labeled antigen method. / Tsuge, Shinya; Mizutani, Yasuyoshi; Matsuoka, Kazuhiro; Sawasaki, Tatsuya; Endo, Yaeta; Naruishi, Koji; Maeda, Hiroshi; Takashiba, Shogo; Shiogama, Kazuya; Inada, Ken ichi; Tsutsumi, Yutaka.

In: Journal of Histochemistry and Cytochemistry, Vol. 59, No. 7, 2011, p. 673-689.

Research output: Contribution to journalArticle

Tsuge, Shinya ; Mizutani, Yasuyoshi ; Matsuoka, Kazuhiro ; Sawasaki, Tatsuya ; Endo, Yaeta ; Naruishi, Koji ; Maeda, Hiroshi ; Takashiba, Shogo ; Shiogama, Kazuya ; Inada, Ken ichi ; Tsutsumi, Yutaka. / Specific in situ visualization of plasma cells producing antibodies against porphyromonas gingivalis in gingival radicular cyst : Application of the enzyme-labeled antigen method. In: Journal of Histochemistry and Cytochemistry. 2011 ; Vol. 59, No. 7. pp. 673-689.
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abstract = "The enzyme-labeled antigen method was applied to visualize plasma cells producing antibodies to Porphyromonas gingivalis, flora of the human oral cavity. Antibodies to P. gingivalis have reportedly been detected in sera of patients with periodontitis. Biotinylated bacterial antigens, Ag53, and four gingipain domains (Arg-pro, Arg-hgp, Lys-pro, and Lys-hgp) were prepared by the cell-free protein synthesis system using the wheat germ extract. In paraformaldehyde-fixed frozen sections of rat lymph nodes experimentally immunized with Ag53-positive and Ag53-negative P. gingivalis, plasma cells were labeled with biotinylated Arg-hgp and Lys-hgp. Antibodies to Ag53 were detected only in the nodes immunized with Ag53-positive bacteria. In two of eight lesions of gingival radicular cyst with inflammatory infiltration, CD138-positive plasma cells in frozen sections were signalized for Arg-hgp and Lys-hgp. An absorption study using unlabeled antigens confirmed the specificity of staining. The AlphaScreen method identified the same-type antibodies in tissue extracts but not in sera. Antibodies to Ag53, Arg-pro, and Lys-pro were undetectable. In two cases, serum antibodies to Arg-hgp and Lys-hgp were AlphaScreen positive, whereas plasma cells were scarcely observed within the lesions. These findings indicate the validity of the enzyme-labeled antigen method. This is the very first application of this novel histochemical technique to human clinical samples.",
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