Human α-enolase from endothelial cells as a target antigen of anti-endothelial cell antibody in Behçet's disease

Kwang Hoon Lee, Hae Shin Chung, Hyoung Sup Kim, Sang Ho Oh, Moon Kyung Ha, Ja-Hyun Baik, Sungnack Lee, Dongsik Bang

Research output: Contribution to journalArticle

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Abstract

Objective. To identify and recombine a protein of the human dermal microvascular endothelial cell (HDMEC) that specifically reacts with anti - endothelial cell antibody (AECA) in the serum of patients with Behçet's disease (BD), and to evaluate the usefulness of this protein in BD. Methods. The proteomics technique, with 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization - time-of-flight (MALDITOF) mass spectrometry, was used to identify and recombine HDMEC antigen. Western blotting and enzyme-linked immunosorbent assay (ELISA) of recombinant protein isolated by gene cloning were performed on serum from healthy controls, patients with BD, and patients with other rheumatic diseases (rheumatoid arthritis, systemic lupus erythematosus, and Wegener's granulomatosis). Results. Eighteen of 40 BD patients had serum IgM antibody to HDMEC antigen. The purified protein that reacted with AECA in BD patient sera was found to be α-enolase by 2-dimensional gel electrophoresis followed by immunoblotting and MALDI-TOF mass spectrometry. Recombinant α-enolase protein was isolated and refined by gene cloning. On Western blots, AECA-positive IgM from the sera of patients with active BD reacted strongly with recombinant human α-enolase. BD patient sera positive for anti - α-enolase did not react with human γ-enolase. On dot-blotting, reactivity to human α-enolase was detected only in the IgM-positive group. Fifteen of the 18 AECA-positive sera that were positive for the HDMEC antigen showed reactivity to recombinant α-enolase IgM antibody by ELISA. Conclusion. The α-enolase protein is the target protein of serum AECA in BD patients. This is the first report of the presence of IgM antibodies to α-enolase in endothelial cells from the serum of BD patients. Although further studies relating this protein to the pathogenesis of BD will be necessary, α-enolase and its antibody may prove useful in the development of new diagnostic and treatment modalities in BD.

Original languageEnglish
Pages (from-to)2025-2035
Number of pages11
JournalArthritis and Rheumatism
Volume48
Issue number7
DOIs
Publication statusPublished - 2003 Jul 1
Externally publishedYes

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Phosphopyruvate Hydratase
Endothelial Cells
Antigens
Immunoglobulin M
Serum
Skin
Antibodies
Proteins
Recombinant Proteins
Electrophoresis
Organism Cloning
Mass Spectrometry
anti-endothelial cell antibody
Western Blotting
Gels
Enzyme-Linked Immunosorbent Assay
Granulomatosis with Polyangiitis
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Rheumatic Diseases
Immunoblotting

ASJC Scopus subject areas

  • Immunology
  • Rheumatology

Cite this

Human α-enolase from endothelial cells as a target antigen of anti-endothelial cell antibody in Behçet's disease. / Lee, Kwang Hoon; Chung, Hae Shin; Kim, Hyoung Sup; Oh, Sang Ho; Ha, Moon Kyung; Baik, Ja-Hyun; Lee, Sungnack; Bang, Dongsik.

In: Arthritis and Rheumatism, Vol. 48, No. 7, 01.07.2003, p. 2025-2035.

Research output: Contribution to journalArticle

Lee, Kwang Hoon ; Chung, Hae Shin ; Kim, Hyoung Sup ; Oh, Sang Ho ; Ha, Moon Kyung ; Baik, Ja-Hyun ; Lee, Sungnack ; Bang, Dongsik. / Human α-enolase from endothelial cells as a target antigen of anti-endothelial cell antibody in Behçet's disease. In: Arthritis and Rheumatism. 2003 ; Vol. 48, No. 7. pp. 2025-2035.
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abstract = "Objective. To identify and recombine a protein of the human dermal microvascular endothelial cell (HDMEC) that specifically reacts with anti - endothelial cell antibody (AECA) in the serum of patients with Beh{\cc}et's disease (BD), and to evaluate the usefulness of this protein in BD. Methods. The proteomics technique, with 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization - time-of-flight (MALDITOF) mass spectrometry, was used to identify and recombine HDMEC antigen. Western blotting and enzyme-linked immunosorbent assay (ELISA) of recombinant protein isolated by gene cloning were performed on serum from healthy controls, patients with BD, and patients with other rheumatic diseases (rheumatoid arthritis, systemic lupus erythematosus, and Wegener's granulomatosis). Results. Eighteen of 40 BD patients had serum IgM antibody to HDMEC antigen. The purified protein that reacted with AECA in BD patient sera was found to be α-enolase by 2-dimensional gel electrophoresis followed by immunoblotting and MALDI-TOF mass spectrometry. Recombinant α-enolase protein was isolated and refined by gene cloning. On Western blots, AECA-positive IgM from the sera of patients with active BD reacted strongly with recombinant human α-enolase. BD patient sera positive for anti - α-enolase did not react with human γ-enolase. On dot-blotting, reactivity to human α-enolase was detected only in the IgM-positive group. Fifteen of the 18 AECA-positive sera that were positive for the HDMEC antigen showed reactivity to recombinant α-enolase IgM antibody by ELISA. Conclusion. The α-enolase protein is the target protein of serum AECA in BD patients. This is the first report of the presence of IgM antibodies to α-enolase in endothelial cells from the serum of BD patients. Although further studies relating this protein to the pathogenesis of BD will be necessary, α-enolase and its antibody may prove useful in the development of new diagnostic and treatment modalities in BD.",
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T1 - Human α-enolase from endothelial cells as a target antigen of anti-endothelial cell antibody in Behçet's disease

AU - Lee, Kwang Hoon

AU - Chung, Hae Shin

AU - Kim, Hyoung Sup

AU - Oh, Sang Ho

AU - Ha, Moon Kyung

AU - Baik, Ja-Hyun

AU - Lee, Sungnack

AU - Bang, Dongsik

PY - 2003/7/1

Y1 - 2003/7/1

N2 - Objective. To identify and recombine a protein of the human dermal microvascular endothelial cell (HDMEC) that specifically reacts with anti - endothelial cell antibody (AECA) in the serum of patients with Behçet's disease (BD), and to evaluate the usefulness of this protein in BD. Methods. The proteomics technique, with 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization - time-of-flight (MALDITOF) mass spectrometry, was used to identify and recombine HDMEC antigen. Western blotting and enzyme-linked immunosorbent assay (ELISA) of recombinant protein isolated by gene cloning were performed on serum from healthy controls, patients with BD, and patients with other rheumatic diseases (rheumatoid arthritis, systemic lupus erythematosus, and Wegener's granulomatosis). Results. Eighteen of 40 BD patients had serum IgM antibody to HDMEC antigen. The purified protein that reacted with AECA in BD patient sera was found to be α-enolase by 2-dimensional gel electrophoresis followed by immunoblotting and MALDI-TOF mass spectrometry. Recombinant α-enolase protein was isolated and refined by gene cloning. On Western blots, AECA-positive IgM from the sera of patients with active BD reacted strongly with recombinant human α-enolase. BD patient sera positive for anti - α-enolase did not react with human γ-enolase. On dot-blotting, reactivity to human α-enolase was detected only in the IgM-positive group. Fifteen of the 18 AECA-positive sera that were positive for the HDMEC antigen showed reactivity to recombinant α-enolase IgM antibody by ELISA. Conclusion. The α-enolase protein is the target protein of serum AECA in BD patients. This is the first report of the presence of IgM antibodies to α-enolase in endothelial cells from the serum of BD patients. Although further studies relating this protein to the pathogenesis of BD will be necessary, α-enolase and its antibody may prove useful in the development of new diagnostic and treatment modalities in BD.

AB - Objective. To identify and recombine a protein of the human dermal microvascular endothelial cell (HDMEC) that specifically reacts with anti - endothelial cell antibody (AECA) in the serum of patients with Behçet's disease (BD), and to evaluate the usefulness of this protein in BD. Methods. The proteomics technique, with 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization - time-of-flight (MALDITOF) mass spectrometry, was used to identify and recombine HDMEC antigen. Western blotting and enzyme-linked immunosorbent assay (ELISA) of recombinant protein isolated by gene cloning were performed on serum from healthy controls, patients with BD, and patients with other rheumatic diseases (rheumatoid arthritis, systemic lupus erythematosus, and Wegener's granulomatosis). Results. Eighteen of 40 BD patients had serum IgM antibody to HDMEC antigen. The purified protein that reacted with AECA in BD patient sera was found to be α-enolase by 2-dimensional gel electrophoresis followed by immunoblotting and MALDI-TOF mass spectrometry. Recombinant α-enolase protein was isolated and refined by gene cloning. On Western blots, AECA-positive IgM from the sera of patients with active BD reacted strongly with recombinant human α-enolase. BD patient sera positive for anti - α-enolase did not react with human γ-enolase. On dot-blotting, reactivity to human α-enolase was detected only in the IgM-positive group. Fifteen of the 18 AECA-positive sera that were positive for the HDMEC antigen showed reactivity to recombinant α-enolase IgM antibody by ELISA. Conclusion. The α-enolase protein is the target protein of serum AECA in BD patients. This is the first report of the presence of IgM antibodies to α-enolase in endothelial cells from the serum of BD patients. Although further studies relating this protein to the pathogenesis of BD will be necessary, α-enolase and its antibody may prove useful in the development of new diagnostic and treatment modalities in BD.

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