Human fibroblasts ubiquitously expressglutamic acid decarboxylase 65 (GAD 65): Possible effects of connective tissue inflammation on gad antibody titer glutamic acid decarboxylase 65 (GAD 65): Possible effects of connective tissue inflammation on GAD antibody titer

Takayuki Kono, Fusanori Nishimura, Hikaru Sugimoto, Kenichi Sikata, Hirofumi Makino, Yoji Murayama

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8 Citations (Scopus)

Abstract

Background: Type 1 diabetes is caused by a destruction of pancreatic β cells due to autoimmunity. Autoantibody against glutamic acid decarboxylase (GAD) 65 expressed in pancreatic β cells is widely used as a predictive marker for pancreatic destruction. In this study, we hypothesized that if certain cells in periodontal tissues could express GAD, then it may influence GAD antibody titer. Methods: We used: 1) reverse transcription-polymerase chain reaction (PCR) analysis to detect GAD 65 mRNA in various cells; 2) nucleotide sequencing analysis to confirm that amplified PCR product is the gene encoding GAD; and 3) Western blotting to determine the expression of GAD 65 protein in human gingival fibroblasts. Immunohistochemical staining of GAD 65 protein in normal and inflamed gingiva was performed to examine the potential influence of periodontal inflammation on GAD 65 expression. GAD antibody titer in sera of periodontal patients as well as healthy subjects was measured to determine if periodontal patients could develop autoantibody against GAD 65. Results: Cultured human gingival, periodontal, and dermal fibroblasts and mesangial cells expressed GAD mRNA. Nucleotide sequencing analyses confirmed the amplified PCR product as GAD 65. Western immunoblotting analyses and immunohistochemical staining revealed that the GAD 65 protein was expressed in vitro and in vivo. The expression of GAD 65 in inflamed tissue was higher than that in normal tissues. Two of 62 periodontal patients without diabetes showed an increased antibody titer against GAD 65, while none of the systemically healthy subjects showed an increased antibody titer against this antigen. Conclusions: We concluded that periodontal inflammation may result in higher levels of GAD and influence GAD antibody titer, and, hence, affect diabetic diagnosis based upon GAD antibody production.

Original languageEnglish
Pages (from-to)598-604
Number of pages7
JournalJournal of Periodontology
Volume72
Issue number5
DOIs
Publication statusPublished - May 2001

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Glutamate Decarboxylase
Carboxy-Lyases
Connective Tissue
Fibroblasts
Inflammation
Acids
Antibodies
Polymerase Chain Reaction
Autoantibodies
Healthy Volunteers
Nucleotides
Western Blotting
Staining and Labeling
Messenger RNA
Proteins
Mesangial Cells
Gingiva

Keywords

  • Diabetes mellitus/diagnosis
  • Glutamic acid decarboxylase 65
  • Inflammatory response
  • Markers, biological
  • Pancreatic diseases/diagnosis
  • Periodontal diseases

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

@article{25fcf8379b0b43069bfe0be2ed7d7552,
title = "Human fibroblasts ubiquitously expressglutamic acid decarboxylase 65 (GAD 65): Possible effects of connective tissue inflammation on gad antibody titer glutamic acid decarboxylase 65 (GAD 65): Possible effects of connective tissue inflammation on GAD antibody titer",
abstract = "Background: Type 1 diabetes is caused by a destruction of pancreatic β cells due to autoimmunity. Autoantibody against glutamic acid decarboxylase (GAD) 65 expressed in pancreatic β cells is widely used as a predictive marker for pancreatic destruction. In this study, we hypothesized that if certain cells in periodontal tissues could express GAD, then it may influence GAD antibody titer. Methods: We used: 1) reverse transcription-polymerase chain reaction (PCR) analysis to detect GAD 65 mRNA in various cells; 2) nucleotide sequencing analysis to confirm that amplified PCR product is the gene encoding GAD; and 3) Western blotting to determine the expression of GAD 65 protein in human gingival fibroblasts. Immunohistochemical staining of GAD 65 protein in normal and inflamed gingiva was performed to examine the potential influence of periodontal inflammation on GAD 65 expression. GAD antibody titer in sera of periodontal patients as well as healthy subjects was measured to determine if periodontal patients could develop autoantibody against GAD 65. Results: Cultured human gingival, periodontal, and dermal fibroblasts and mesangial cells expressed GAD mRNA. Nucleotide sequencing analyses confirmed the amplified PCR product as GAD 65. Western immunoblotting analyses and immunohistochemical staining revealed that the GAD 65 protein was expressed in vitro and in vivo. The expression of GAD 65 in inflamed tissue was higher than that in normal tissues. Two of 62 periodontal patients without diabetes showed an increased antibody titer against GAD 65, while none of the systemically healthy subjects showed an increased antibody titer against this antigen. Conclusions: We concluded that periodontal inflammation may result in higher levels of GAD and influence GAD antibody titer, and, hence, affect diabetic diagnosis based upon GAD antibody production.",
keywords = "Diabetes mellitus/diagnosis, Glutamic acid decarboxylase 65, Inflammatory response, Markers, biological, Pancreatic diseases/diagnosis, Periodontal diseases",
author = "Takayuki Kono and Fusanori Nishimura and Hikaru Sugimoto and Kenichi Sikata and Hirofumi Makino and Yoji Murayama",
year = "2001",
month = "5",
doi = "10.1902/jop.2001.72.5.598",
language = "English",
volume = "72",
pages = "598--604",
journal = "Journal of Periodontology",
issn = "0022-3492",
publisher = "American Academy of Periodontology",
number = "5",

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TY - JOUR

T1 - Human fibroblasts ubiquitously expressglutamic acid decarboxylase 65 (GAD 65)

T2 - Possible effects of connective tissue inflammation on gad antibody titer glutamic acid decarboxylase 65 (GAD 65): Possible effects of connective tissue inflammation on GAD antibody titer

AU - Kono, Takayuki

AU - Nishimura, Fusanori

AU - Sugimoto, Hikaru

AU - Sikata, Kenichi

AU - Makino, Hirofumi

AU - Murayama, Yoji

PY - 2001/5

Y1 - 2001/5

N2 - Background: Type 1 diabetes is caused by a destruction of pancreatic β cells due to autoimmunity. Autoantibody against glutamic acid decarboxylase (GAD) 65 expressed in pancreatic β cells is widely used as a predictive marker for pancreatic destruction. In this study, we hypothesized that if certain cells in periodontal tissues could express GAD, then it may influence GAD antibody titer. Methods: We used: 1) reverse transcription-polymerase chain reaction (PCR) analysis to detect GAD 65 mRNA in various cells; 2) nucleotide sequencing analysis to confirm that amplified PCR product is the gene encoding GAD; and 3) Western blotting to determine the expression of GAD 65 protein in human gingival fibroblasts. Immunohistochemical staining of GAD 65 protein in normal and inflamed gingiva was performed to examine the potential influence of periodontal inflammation on GAD 65 expression. GAD antibody titer in sera of periodontal patients as well as healthy subjects was measured to determine if periodontal patients could develop autoantibody against GAD 65. Results: Cultured human gingival, periodontal, and dermal fibroblasts and mesangial cells expressed GAD mRNA. Nucleotide sequencing analyses confirmed the amplified PCR product as GAD 65. Western immunoblotting analyses and immunohistochemical staining revealed that the GAD 65 protein was expressed in vitro and in vivo. The expression of GAD 65 in inflamed tissue was higher than that in normal tissues. Two of 62 periodontal patients without diabetes showed an increased antibody titer against GAD 65, while none of the systemically healthy subjects showed an increased antibody titer against this antigen. Conclusions: We concluded that periodontal inflammation may result in higher levels of GAD and influence GAD antibody titer, and, hence, affect diabetic diagnosis based upon GAD antibody production.

AB - Background: Type 1 diabetes is caused by a destruction of pancreatic β cells due to autoimmunity. Autoantibody against glutamic acid decarboxylase (GAD) 65 expressed in pancreatic β cells is widely used as a predictive marker for pancreatic destruction. In this study, we hypothesized that if certain cells in periodontal tissues could express GAD, then it may influence GAD antibody titer. Methods: We used: 1) reverse transcription-polymerase chain reaction (PCR) analysis to detect GAD 65 mRNA in various cells; 2) nucleotide sequencing analysis to confirm that amplified PCR product is the gene encoding GAD; and 3) Western blotting to determine the expression of GAD 65 protein in human gingival fibroblasts. Immunohistochemical staining of GAD 65 protein in normal and inflamed gingiva was performed to examine the potential influence of periodontal inflammation on GAD 65 expression. GAD antibody titer in sera of periodontal patients as well as healthy subjects was measured to determine if periodontal patients could develop autoantibody against GAD 65. Results: Cultured human gingival, periodontal, and dermal fibroblasts and mesangial cells expressed GAD mRNA. Nucleotide sequencing analyses confirmed the amplified PCR product as GAD 65. Western immunoblotting analyses and immunohistochemical staining revealed that the GAD 65 protein was expressed in vitro and in vivo. The expression of GAD 65 in inflamed tissue was higher than that in normal tissues. Two of 62 periodontal patients without diabetes showed an increased antibody titer against GAD 65, while none of the systemically healthy subjects showed an increased antibody titer against this antigen. Conclusions: We concluded that periodontal inflammation may result in higher levels of GAD and influence GAD antibody titer, and, hence, affect diabetic diagnosis based upon GAD antibody production.

KW - Diabetes mellitus/diagnosis

KW - Glutamic acid decarboxylase 65

KW - Inflammatory response

KW - Markers, biological

KW - Pancreatic diseases/diagnosis

KW - Periodontal diseases

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