Specific In Situ Visualization of Plasma Cells Producing Antibodies against Porphyromonas gingivalis in Gingival Radicular Cyst

Author:

Tsuge Shinya12345,Mizutani Yasuyoshi12345,Matsuoka Kazuhiro12345,Sawasaki Tatsuya12345,Endo Yaeta12345,Naruishi Koji12345,Maeda Hiroshi12345,Takashiba Shogo12345,Shiogama Kazuya12345,Inada Ken-ichi12345,Tsutsumi Yutaka12345

Affiliation:

1. Department of Pathology (STsuge,YM,KS,KI,YT), Fujita Health University School of Medicine, Toyoake, Aichi, Japan

2. Department of Oral and Maxillofacial Surgery (STsuge), Fujita Health University School of Medicine, Toyoake, Aichi, Japan

3. Cell-Free Science and Technology Research Center, Ehime University, Matsuyama, Japan (KM,TS,YE)

4. Proteo-Medicine Research Center, Ehime University, Toon, Japan (TS,YE)

5. Department of Pathophysiology–Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan (KN,HM,STakashiba)

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.

Publisher

SAGE Publications

Subject

Histology,Anatomy

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