Comparative analysis of novel autoantibody isotypes against citrullinated-inter-alpha-trypsin inhibitor heavy chain 3 (ITIH3)542-556 peptide in serum from Taiwanese females with rheumatoid arthritis, primary Sjögren's syndrome and secondary Sjögren's syndrome in rheumatoid arthritis

Chen Chung Liao, Pei Lun Chou, Chao-Wen Cheng, Yu Sheng Chang, Wei Ming Chi, Kai Leun Tsai, Wei Jung Chen, Ting Shuan Kung, Chih Chun Tai, Kuan Wei Lee, You Chia Chen, Ching Yu Lin

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Abstract

The purpose of this study was to discover and validate inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3) as novel biomarkers, and evaluate autoantibody isotypes against an unmodified and citrullinated ITIH3542-556 peptide among Taiwanese female patients with rheumatoid arthritis (RA), primary Sjögren's syndrome (pSS), secondary Sjögren's syndrome in rheumatoid arthritis (RA-sSS), and healthy controls (HCs). We used concanavalin A (Con A) affinity chromatography, 1-D SDS-PAGE, and label-free nano-LC-MS/MS to screen biomarker candidates (serum-derived Con A-captured proteins) and then identify PTMs of validated biomarkers (serum proteins) using pooled serum from 7 RA-sSS female patients and 7 age-matched HCs (the discovery set). Furthermore, the protein level and autoantibody isotype analyses were further validated against individual serum from 18 HCs, 18 RA, 18 pSS, and 18 RA-sSS patients (the validation set). Con A-bound ITIH3 was identified and validated as the only differentially expressed protein, which was elevated. Additionally, 2 novel PTMs in ITIH3 were identified and included citrullination at arginine-546 and arginine-556, and hexosamine at tryptophan-558. Further, concentrations of anti-citrullinatd-ITIH3542-556 peptide autoantibodies significantly increased in patients with RA, pSS, and RA-sSS compared to HCs. Especially, autoantibody IgM against the citrullinated-ITIH3542-556 peptide showed better diagnostic performance in discriminating both RA versus pSS and pSS versus RA-sSS. Significance: By using comparative proteomic analysis of serum samples, the current study discovered and validates differentially expressed Con A-bound ITIH3 as a potential biomarker for secondary Sjögren's syndrome (SS) in rheumatoid arthritis (RA) patients and healthy controls (HCs). Besides, hexosamine and citrullination on ITIH3 were further identified. Through analyzing autoantibody isotypes against the citrullinated ITIH3 peptide, patients with RA, primary SS, and RA-secondary SS, and HCs can be further discriminated. The current strategy can be applied for identifying potential diagnostic and pathologic markers for autoimmune diseases.

Original languageEnglish
JournalJournal of Proteomics
Volume141
DOIs
Publication statusPublished - Jun 1 2016

Fingerprint

Autoantibodies
Rheumatoid Arthritis
Biomarkers
Concanavalin A
Peptides
Serum
Pulse time modulation
Hexosamines
Arginine
Affinity chromatography
Proteins
Tryptophan
Immunoglobulin M
inter-alpha-inhibitor
Labels
Blood Proteins
Affinity Chromatography
Proteomics
Autoimmune Diseases
Polyacrylamide Gel Electrophoresis

Keywords

  • Autoantibody isotypes
  • Citrullination
  • Primary Sjögren's syndrome
  • Rheumatoid arthritis
  • Secondary Sjögren's syndrome in rheumatoid arthritis
  • Serum

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

@article{190663dbfe5d400ab32c4bf709aa0663,
title = "Comparative analysis of novel autoantibody isotypes against citrullinated-inter-alpha-trypsin inhibitor heavy chain 3 (ITIH3)542-556 peptide in serum from Taiwanese females with rheumatoid arthritis, primary Sj{\"o}gren's syndrome and secondary Sj{\"o}gren's syndrome in rheumatoid arthritis",
abstract = "The purpose of this study was to discover and validate inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3) as novel biomarkers, and evaluate autoantibody isotypes against an unmodified and citrullinated ITIH3542-556 peptide among Taiwanese female patients with rheumatoid arthritis (RA), primary Sj{\"o}gren's syndrome (pSS), secondary Sj{\"o}gren's syndrome in rheumatoid arthritis (RA-sSS), and healthy controls (HCs). We used concanavalin A (Con A) affinity chromatography, 1-D SDS-PAGE, and label-free nano-LC-MS/MS to screen biomarker candidates (serum-derived Con A-captured proteins) and then identify PTMs of validated biomarkers (serum proteins) using pooled serum from 7 RA-sSS female patients and 7 age-matched HCs (the discovery set). Furthermore, the protein level and autoantibody isotype analyses were further validated against individual serum from 18 HCs, 18 RA, 18 pSS, and 18 RA-sSS patients (the validation set). Con A-bound ITIH3 was identified and validated as the only differentially expressed protein, which was elevated. Additionally, 2 novel PTMs in ITIH3 were identified and included citrullination at arginine-546 and arginine-556, and hexosamine at tryptophan-558. Further, concentrations of anti-citrullinatd-ITIH3542-556 peptide autoantibodies significantly increased in patients with RA, pSS, and RA-sSS compared to HCs. Especially, autoantibody IgM against the citrullinated-ITIH3542-556 peptide showed better diagnostic performance in discriminating both RA versus pSS and pSS versus RA-sSS. Significance: By using comparative proteomic analysis of serum samples, the current study discovered and validates differentially expressed Con A-bound ITIH3 as a potential biomarker for secondary Sj{\"o}gren's syndrome (SS) in rheumatoid arthritis (RA) patients and healthy controls (HCs). Besides, hexosamine and citrullination on ITIH3 were further identified. Through analyzing autoantibody isotypes against the citrullinated ITIH3 peptide, patients with RA, primary SS, and RA-secondary SS, and HCs can be further discriminated. The current strategy can be applied for identifying potential diagnostic and pathologic markers for autoimmune diseases.",
keywords = "Autoantibody isotypes, Citrullination, Primary Sj{\"o}gren's syndrome, Rheumatoid arthritis, Secondary Sj{\"o}gren's syndrome in rheumatoid arthritis, Serum",
author = "Liao, {Chen Chung} and Chou, {Pei Lun} and Chao-Wen Cheng and Chang, {Yu Sheng} and Chi, {Wei Ming} and Tsai, {Kai Leun} and Chen, {Wei Jung} and Kung, {Ting Shuan} and Tai, {Chih Chun} and Lee, {Kuan Wei} and Chen, {You Chia} and Lin, {Ching Yu}",
year = "2016",
month = "6",
day = "1",
doi = "10.1016/j.jprot.2016.03.031",
language = "English",
volume = "141",
journal = "Journal of Proteomics",
issn = "1874-3919",
publisher = "Elsevier",

}

TY - JOUR

T1 - Comparative analysis of novel autoantibody isotypes against citrullinated-inter-alpha-trypsin inhibitor heavy chain 3 (ITIH3)542-556 peptide in serum from Taiwanese females with rheumatoid arthritis, primary Sjögren's syndrome and secondary Sjögren's syndrome in rheumatoid arthritis

AU - Liao, Chen Chung

AU - Chou, Pei Lun

AU - Cheng, Chao-Wen

AU - Chang, Yu Sheng

AU - Chi, Wei Ming

AU - Tsai, Kai Leun

AU - Chen, Wei Jung

AU - Kung, Ting Shuan

AU - Tai, Chih Chun

AU - Lee, Kuan Wei

AU - Chen, You Chia

AU - Lin, Ching Yu

PY - 2016/6/1

Y1 - 2016/6/1

N2 - The purpose of this study was to discover and validate inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3) as novel biomarkers, and evaluate autoantibody isotypes against an unmodified and citrullinated ITIH3542-556 peptide among Taiwanese female patients with rheumatoid arthritis (RA), primary Sjögren's syndrome (pSS), secondary Sjögren's syndrome in rheumatoid arthritis (RA-sSS), and healthy controls (HCs). We used concanavalin A (Con A) affinity chromatography, 1-D SDS-PAGE, and label-free nano-LC-MS/MS to screen biomarker candidates (serum-derived Con A-captured proteins) and then identify PTMs of validated biomarkers (serum proteins) using pooled serum from 7 RA-sSS female patients and 7 age-matched HCs (the discovery set). Furthermore, the protein level and autoantibody isotype analyses were further validated against individual serum from 18 HCs, 18 RA, 18 pSS, and 18 RA-sSS patients (the validation set). Con A-bound ITIH3 was identified and validated as the only differentially expressed protein, which was elevated. Additionally, 2 novel PTMs in ITIH3 were identified and included citrullination at arginine-546 and arginine-556, and hexosamine at tryptophan-558. Further, concentrations of anti-citrullinatd-ITIH3542-556 peptide autoantibodies significantly increased in patients with RA, pSS, and RA-sSS compared to HCs. Especially, autoantibody IgM against the citrullinated-ITIH3542-556 peptide showed better diagnostic performance in discriminating both RA versus pSS and pSS versus RA-sSS. Significance: By using comparative proteomic analysis of serum samples, the current study discovered and validates differentially expressed Con A-bound ITIH3 as a potential biomarker for secondary Sjögren's syndrome (SS) in rheumatoid arthritis (RA) patients and healthy controls (HCs). Besides, hexosamine and citrullination on ITIH3 were further identified. Through analyzing autoantibody isotypes against the citrullinated ITIH3 peptide, patients with RA, primary SS, and RA-secondary SS, and HCs can be further discriminated. The current strategy can be applied for identifying potential diagnostic and pathologic markers for autoimmune diseases.

AB - The purpose of this study was to discover and validate inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3) as novel biomarkers, and evaluate autoantibody isotypes against an unmodified and citrullinated ITIH3542-556 peptide among Taiwanese female patients with rheumatoid arthritis (RA), primary Sjögren's syndrome (pSS), secondary Sjögren's syndrome in rheumatoid arthritis (RA-sSS), and healthy controls (HCs). We used concanavalin A (Con A) affinity chromatography, 1-D SDS-PAGE, and label-free nano-LC-MS/MS to screen biomarker candidates (serum-derived Con A-captured proteins) and then identify PTMs of validated biomarkers (serum proteins) using pooled serum from 7 RA-sSS female patients and 7 age-matched HCs (the discovery set). Furthermore, the protein level and autoantibody isotype analyses were further validated against individual serum from 18 HCs, 18 RA, 18 pSS, and 18 RA-sSS patients (the validation set). Con A-bound ITIH3 was identified and validated as the only differentially expressed protein, which was elevated. Additionally, 2 novel PTMs in ITIH3 were identified and included citrullination at arginine-546 and arginine-556, and hexosamine at tryptophan-558. Further, concentrations of anti-citrullinatd-ITIH3542-556 peptide autoantibodies significantly increased in patients with RA, pSS, and RA-sSS compared to HCs. Especially, autoantibody IgM against the citrullinated-ITIH3542-556 peptide showed better diagnostic performance in discriminating both RA versus pSS and pSS versus RA-sSS. Significance: By using comparative proteomic analysis of serum samples, the current study discovered and validates differentially expressed Con A-bound ITIH3 as a potential biomarker for secondary Sjögren's syndrome (SS) in rheumatoid arthritis (RA) patients and healthy controls (HCs). Besides, hexosamine and citrullination on ITIH3 were further identified. Through analyzing autoantibody isotypes against the citrullinated ITIH3 peptide, patients with RA, primary SS, and RA-secondary SS, and HCs can be further discriminated. The current strategy can be applied for identifying potential diagnostic and pathologic markers for autoimmune diseases.

KW - Autoantibody isotypes

KW - Citrullination

KW - Primary Sjögren's syndrome

KW - Rheumatoid arthritis

KW - Secondary Sjögren's syndrome in rheumatoid arthritis

KW - Serum

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U2 - 10.1016/j.jprot.2016.03.031

DO - 10.1016/j.jprot.2016.03.031

M3 - Article

C2 - 27072115

AN - SCOPUS:84964318556

VL - 141

JO - Journal of Proteomics

JF - Journal of Proteomics

SN - 1874-3919

ER -