TY - JOUR
T1 - Application of synthetic peptides for detection of anti-citrullinated peptide antibodies
AU - Trier, Nicole Hartwig
AU - Holm, Bettina Eide
AU - Slot, Ole
AU - Locht, Henning
AU - Lindegaard, Hanne
AU - Svendsen, Anders
AU - Nielsen, Christoffer Tandrup
AU - Jacobsen, Søren
AU - Theander, Elke
AU - Houen, Gunnar
N1 - Copyright © 2016 Elsevier Inc. All rights reserved.
PY - 2016/2/1
Y1 - 2016/2/1
N2 - Anti-citrullinated protein antibodies (ACPAs) are a hallmark of rheumatoid arthritis (RA) and represent an important tool for the serological diagnosis of RA. In this study, we describe ACPA reactivity to overlapping citrullinated Epstein-Barr virus nuclear antigen-1 (EBNA-1)-derived peptides and analyze their potential as substrates for ACPA detection by streptavidin capture enzyme-linked immunosorbent assay. Using systematically overlapping peptides, containing a 10 amino acid overlap, labelled with biotin C-terminally or N-terminally, sera from 160 individuals (RA sera (n=60), healthy controls (n=40), systemic lupus erythematosus (n=20), Sjögren's syndrome (n=40)) were screened for antibody reactivity. Antibodies to a panel of five citrullinated EBNA-1 peptides were found in 67% of RA sera, exclusively of the IgG isotype, while 53% of the patient sera reacted with a single peptide, ARGGSRERARGRGRG-Cit-GEKR, accounting for more than half of the ACPA reactivity alone. Moreover, these antibodies were detected in 10% of CCP2-negative RA sera. In addition, 47% of the RA sera reacted with two or three citrullinated EBNA-1 peptides from the selected peptide panel. Furthermore, a negative correlation between the biotin attachment site and the location of citrulline in the peptides was found, i.e. the closer the citrulline was located to biotin, the lower the antibody reactivity. Our data suggest that citrullinated EBNA-1 peptides may be considered a substrate for the detection of ACPAs and that the presence of Epstein-Barr virus may play a role in the induction of these autoantibodies.
AB - Anti-citrullinated protein antibodies (ACPAs) are a hallmark of rheumatoid arthritis (RA) and represent an important tool for the serological diagnosis of RA. In this study, we describe ACPA reactivity to overlapping citrullinated Epstein-Barr virus nuclear antigen-1 (EBNA-1)-derived peptides and analyze their potential as substrates for ACPA detection by streptavidin capture enzyme-linked immunosorbent assay. Using systematically overlapping peptides, containing a 10 amino acid overlap, labelled with biotin C-terminally or N-terminally, sera from 160 individuals (RA sera (n=60), healthy controls (n=40), systemic lupus erythematosus (n=20), Sjögren's syndrome (n=40)) were screened for antibody reactivity. Antibodies to a panel of five citrullinated EBNA-1 peptides were found in 67% of RA sera, exclusively of the IgG isotype, while 53% of the patient sera reacted with a single peptide, ARGGSRERARGRGRG-Cit-GEKR, accounting for more than half of the ACPA reactivity alone. Moreover, these antibodies were detected in 10% of CCP2-negative RA sera. In addition, 47% of the RA sera reacted with two or three citrullinated EBNA-1 peptides from the selected peptide panel. Furthermore, a negative correlation between the biotin attachment site and the location of citrulline in the peptides was found, i.e. the closer the citrulline was located to biotin, the lower the antibody reactivity. Our data suggest that citrullinated EBNA-1 peptides may be considered a substrate for the detection of ACPAs and that the presence of Epstein-Barr virus may play a role in the induction of these autoantibodies.
KW - Amino Acid Sequence
KW - Antibody Specificity
KW - Arthritis, Rheumatoid
KW - Autoantibodies
KW - Case-Control Studies
KW - Citrulline
KW - Herpesvirus 4, Human
KW - Humans
KW - Lupus Erythematosus, Systemic
KW - Molecular Sequence Data
KW - Peptide Fragments
KW - Peptides
KW - Viral Proteins
KW - Journal Article
U2 - 10.1016/j.peptides.2016.01.005
DO - 10.1016/j.peptides.2016.01.005
M3 - Journal article
C2 - 26796582
SN - 0196-9781
VL - 76
SP - 87
EP - 95
JO - Peptides
JF - Peptides
ER -