High-Titer Rheumatoid Arthritis Antibodies Preferentially Bind Fibrinogen Citrullinated by Peptidylarginine Deiminase 4.
High-Titer Rheumatoid Arthritis Antibodies Preferentially Bind Fibrinogen Citrullinated by Peptidylarginine Deiminase 4.
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DOI：
10.1002/art.40035
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发表时间：
2017-05
期刊：
影响因子：
--
通讯作者：
Orange DE
中科院分区：
文献类型：
--
作者：
Blachère NE;Parveen S;Frank MO;Dill BD;Molina H;Orange DE
大多数类风湿性关节炎（RA）患者携带瓜氨酸化自身抗原抗体，如瓜氨酸化纤维蛋白原。在RA滑膜中检测到两种不同底物特异性催化瓜氨酸化的肽基精氨酸脱亚胺酶（PAD2和PAD4）同工型。本研究旨在确定RA患者的抗体是否优先结合PAD2或PAD4瓜氨酸纤维蛋白原。采用ELISA和Western blot检测RA患者和正常供体血浆与不同稀释度的PAD2或PAD4瓜氨酸纤维蛋白原、天然纤维蛋白原或瓜氨酸纤维蛋白原肽的结合情况。通过Western blot检测显示RA患者反应性的质量对应的条带被切除并通过质谱分析。在低滴度下，RA患者对PAD2和PAD4瓜氨酸纤维蛋白原的抗体反应性无显著差异。当血浆进一步稀释至1:250和1:1000时，通过ELISA和Western blot检测，RA患者血浆中结合的PAD4瓜氨酸化纤维蛋白原明显多于PAD2瓜氨酸化纤维蛋白原。抗体滴度的增加与PAD2或PAD4瓜氨酸纤维蛋白原的亲和力增加有关。这两种酶都能高瓜氨酸化纤维蛋白原，但PAD4能间歇性地高瓜氨酸化精氨酸，产生瓜氨酸化和非修饰精氨酸的混合物。多肽ELISA和预吸附实验证实，间歇性瓜氨酸化区域占RA抗体结合瓜氨酸化纤维蛋白原β链的大部分。在高滴度下，RA患者抗体优先结合PAD4修饰的纤维蛋白原，因为间歇性瓜氨酸化提供了更多样化的瓜氨酸化表位。
Most patients with rheumatoid arthritis (RA) harbor antibodies to citrullinated autoantigens, such as citrullinated fibrinogen. Two isoforms of peptidyl arginine deiminase (PAD), PAD2 and PAD4, which catalyze citrullination with different substrate specificities, can be detected in RA synovium. This study was undertaken to determine whether RA patient antibodies preferentially bind PAD2 or PAD4 citrullinated fibrinogen. RA patient and normal donor plasma was tested for binding to PAD2 or PAD4 citrullinated fibrinogen, native fibrinogen, or citrullinated fibrinogen peptides in various dilutions by ELISA and Western blot. Bands corresponding to masses demonstrating RA patient reactivity by Western blot were excised and analyzed by mass spectrometry. At low titer, there was no significant difference between RA patient antibody reactivity to PAD2 and PAD4 citrullinated fibrinogen. When plasma was further diluted to 1:250 and 1:1000, RA patient plasma bound PAD4 citrullinated fibrinogen significantly more than PAD2 citrullinated fibrinogen, as measured by ELISA and Western blot. Increased antibody titer was associated with increased avidity for both PAD2 or PAD4 citrullinated fibrinogen. Both enzymes hypercitrullinated fibrinogen, but PAD4 citrullinates arginines more intermittently, generating a mix of citrullinated and nonmodifed arginines. Peptide ELISA and preadsorption assays confirmed that the region of intermittent citrullination accounts for the majority of RA antibody binding to the beta chain of citrullinated fibrinogen. At high titer, RA patient antibodies preferentially bind fibrinogen modified by PAD4 because intermittent citrullination offers a more diverse assortment of citrullinated epitopes.
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DOI：
10.1084/jem.20100239
发表时间：
2010-08-30
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Li P;Li M;Lindberg MR;Kennett MJ;Xiong N;Wang Y
通讯作者：
Wang Y
影响因子：
--
作者：
Snir, Omri;Widhe, Mona;Malmstrom, Vivianne
通讯作者：
Malmstrom, Vivianne
影响因子：
4.9
作者：
Blachère NE;Parveen S;Fak J;Frank MO;Orange DE
通讯作者：
Orange DE
影响因子：
4.9
作者：
Mjaavatten MD;van der Heijde D;Uhlig T;Haugen AJ;Nygaard H;Sidenvall G;Helgetveit K;Kvien TK
通讯作者：
Kvien TK
DOI：
10.1002/art.39313
发表时间：
2015-12
期刊：
Arthritis & rheumatology (Hoboken, N.J.)
影响因子：
--
作者：
Spengler J;Lugonja B;Ytterberg AJ;Zubarev RA;Creese AJ;Pearson MJ;Grant MM;Milward M;Lundberg K;Buckley CD;Filer A;Raza K;Cooper PR;Chapple IL;Scheel-Toellner D
通讯作者：
Scheel-Toellner D