Enzyme-labeled antigen method: Development and application of the novel approach for identifying plasma cells locally producing disease-specific antibodies in inflammatory lesions

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1 Citation (Scopus)

Abstract

In chronic inflammatory lesions of autoimmune and infectious diseases, plasma cells are frequently observed. Antigens recognized by antibodies produced by the plasma cells mostly remain unclear. A new technique identifying these corresponding antigens may give us a breakthrough for understanding the disease from a pathophysiological viewpoint, simply because the immunocytes are seen within the lesion. We have developed an enzymelabeled antigen method for microscopic identification of the antigen recognized by specific antibodies locally produced in plasma cells in inflammatory lesions. Firstly, target biotinylated antigens were constructed by the wheat germ cell-free protein synthesis system or through chemical biotinylation. Next, proteins reactive to antibodies in tissue extracts were screened and antibody titers were evaluated by the AlphaScreen method. Finally, with the enzymelabeled antigen method using the biotinylated antigens as probes, plasma cells producing specific antibodies were microscopically localized in fixed frozen sections. Our novel approach visualized tissue plasma cells that produced 1) autoantibodies in rheumatoid arthritis, 2) antibodies against major antigens of Porphyromonas gingivalis in periodontitis or radicular cyst, and 3) antibodies against a carbohydrate antigen, Strep A, of Streptococcus pyogenes in recurrent tonsillitis. Evaluation of local specific antibody responses expectedly contributes to clarifying previously unknown processes in inflammatory disorders.

Original languageEnglish
Pages (from-to)7-19
Number of pages13
JournalActa Histochemica et Cytochemica
Volume49
Issue number1
DOIs
Publication statusPublished - 27-02-2016

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Plasma Cells
Plasmas
Antigens
Antibodies
Enzymes
Plasma probes
Radicular Cyst
Biotinylation
Tonsillitis
Porphyromonas gingivalis
Tissue Extracts
Streptococcus pyogenes
Periodontitis
Frozen Sections
Germ Cells
Autoantibodies
Autoimmune Diseases
Triticum
Antibody Formation
Communicable Diseases

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Biochemistry
  • Physiology
  • Histology
  • Cell Biology

Cite this

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abstract = "In chronic inflammatory lesions of autoimmune and infectious diseases, plasma cells are frequently observed. Antigens recognized by antibodies produced by the plasma cells mostly remain unclear. A new technique identifying these corresponding antigens may give us a breakthrough for understanding the disease from a pathophysiological viewpoint, simply because the immunocytes are seen within the lesion. We have developed an enzymelabeled antigen method for microscopic identification of the antigen recognized by specific antibodies locally produced in plasma cells in inflammatory lesions. Firstly, target biotinylated antigens were constructed by the wheat germ cell-free protein synthesis system or through chemical biotinylation. Next, proteins reactive to antibodies in tissue extracts were screened and antibody titers were evaluated by the AlphaScreen method. Finally, with the enzymelabeled antigen method using the biotinylated antigens as probes, plasma cells producing specific antibodies were microscopically localized in fixed frozen sections. Our novel approach visualized tissue plasma cells that produced 1) autoantibodies in rheumatoid arthritis, 2) antibodies against major antigens of Porphyromonas gingivalis in periodontitis or radicular cyst, and 3) antibodies against a carbohydrate antigen, Strep A, of Streptococcus pyogenes in recurrent tonsillitis. Evaluation of local specific antibody responses expectedly contributes to clarifying previously unknown processes in inflammatory disorders.",
author = "Yasuyoshi Mizutani and Kazuya Shiogama and Takanori Onouchi and Kouhei Sakurai and Inada, {Ken Ichi} and Yutaka Tsutsumi",
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AU - Mizutani, Yasuyoshi

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AU - Onouchi, Takanori

AU - Sakurai, Kouhei

AU - Inada, Ken Ichi

AU - Tsutsumi, Yutaka

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