In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis

The application of the enzyme-labeled antigen method

Yasuyoshi Mizutani, S. Tsuge, H. Takeda, Y. Hasegawa, Kazuya Shiogama, Takanori Onouchi, K. Inada, T. Sawasaki, Y. Tsutsumi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Porphyromonas gingivalis is a keystone periodontal pathogen. Histologocally, the gingival tissue in periodontitis shows dense infiltration of plasma cells. However, antigens recognized by antibodies secreted from the immunocytes remain unknown. The enzyme-labeled antigen method was applied to detecting plasma cells producing P. gingivalis-specific antibodies in biopsied gingival tissue of periodontitis. N-terminally biotinylated P. gingivalis 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 wheatgerm extract. With these five labeled proteins as probes, 20 lesions of periodontitis were evaluated. With the AlphaScreen method, antibodies against any one of the five P. gingivalis antigens were detected in 11 (55%) serum samples and 17 (85%) tissue extracts. Using the enzyme-labeled antigen method on paraformaldehyde-fixed frozen sections of gingival tissue, plasma cells were labeled with any one of the five antigens in 17 (94%) of 18 specimens, in which evaluable plasma cells were detected. The positivity rates in periodontitis were significantly higher than those found previously in radicular cysts (20% in sera and 33% in tissue extracts with the AlphaScreen method, and 25% with the enzyme-labeled antigen method). Our findings directly indicate that antibodies reactive to P. gingivalis are locally produced in the gingival lesions, and that inflammatory reactions against P. gingivalis are involved in periodontitis.

Original languageEnglish
Pages (from-to)156-173
Number of pages18
JournalMolecular Oral Microbiology
Volume29
Issue number4
DOIs
Publication statusPublished - 01-01-2014

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Porphyromonas gingivalis
Periodontitis
Plasma Cells
Antigens
Antibodies
Enzymes
Tissue Extracts
arginylproline
lysylproline
Radicular Cyst
Frozen Sections
Serum
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Dentistry(all)
  • Microbiology (medical)

Cite this

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title = "In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis: The application of the enzyme-labeled antigen method",
abstract = "Porphyromonas gingivalis is a keystone periodontal pathogen. Histologocally, the gingival tissue in periodontitis shows dense infiltration of plasma cells. However, antigens recognized by antibodies secreted from the immunocytes remain unknown. The enzyme-labeled antigen method was applied to detecting plasma cells producing P. gingivalis-specific antibodies in biopsied gingival tissue of periodontitis. N-terminally biotinylated P. gingivalis 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 wheatgerm extract. With these five labeled proteins as probes, 20 lesions of periodontitis were evaluated. With the AlphaScreen method, antibodies against any one of the five P. gingivalis antigens were detected in 11 (55{\%}) serum samples and 17 (85{\%}) tissue extracts. Using the enzyme-labeled antigen method on paraformaldehyde-fixed frozen sections of gingival tissue, plasma cells were labeled with any one of the five antigens in 17 (94{\%}) of 18 specimens, in which evaluable plasma cells were detected. The positivity rates in periodontitis were significantly higher than those found previously in radicular cysts (20{\%} in sera and 33{\%} in tissue extracts with the AlphaScreen method, and 25{\%} with the enzyme-labeled antigen method). Our findings directly indicate that antibodies reactive to P. gingivalis are locally produced in the gingival lesions, and that inflammatory reactions against P. gingivalis are involved in periodontitis.",
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In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis : The application of the enzyme-labeled antigen method. / Mizutani, Yasuyoshi; Tsuge, S.; Takeda, H.; Hasegawa, Y.; Shiogama, Kazuya; Onouchi, Takanori; Inada, K.; Sawasaki, T.; Tsutsumi, Y.

In: Molecular Oral Microbiology, Vol. 29, No. 4, 01.01.2014, p. 156-173.

Research output: Contribution to journalArticle

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AU - Mizutani, Yasuyoshi

AU - Tsuge, S.

AU - Takeda, H.

AU - Hasegawa, Y.

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AU - Tsutsumi, Y.

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