Cross-reactivity of anti-modified protein antibodies is also present in predisease and individuals without rheumatoid arthritis
Cross-reactivity of anti-modified protein antibodies is also present in predisease and individuals without rheumatoid arthritis
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抗修饰蛋白抗体的交叉反应性也存在于疾病前期和没有类风湿性关节炎的个体中
DOI：
10.1136/annrheumdis-2022-222326
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发表时间：
2022
影响因子：
27.4
通讯作者：
Toes Rene EM
中科院分区：
文献类型：
--
作者：
Reijm Sanne;Brehler Astrid S;Rantapaa-Dahlqvist Solbritt;Kawakami Atsushi;Maeda Takahiro;Kawashiri Shin-ya;Tamai Mami;van der Woude Diane;Toes Rene EM
图1 (A)使用CCP4、CHcitP4和CAcetylP4肽作为抗原时，19例RA发病前患者的ACPA、anti-CarP和AAPA IgG水平随时间变化的任意单位/ ml （aU/ ml）。左图显示发病前数年的数据。右边的热图显示了每个时间点AMPA IgG水平的汇总。(B)日本ACPA-非RA样本（n= 197）、日本ACPA +非RA样本（n= 54）、荷兰健康供者（n= 30）和确诊RA患者（n= 29）的aU/ml中ACPA、anti-CarP和AAPA IgG水平。(C)日本ACPA+非RA样品中ACPA、anti-CarP和AAPA水平aU/mL的相关性。R=相关系数。(D)用CCP4、CHcitP4或CAcetylP4肽分离日本ACPA +非RA样品后，6个样品的ACPA、anti-CarP和AAPA IgG在aU/mL中的水平。抗硝化蛋白抗体；抗乙酰化蛋白抗体；抗carpa，抗氨甲酰化蛋白抗体；RA，类风湿性关节炎。在单克隆和多克隆抗体水平上均显示为乙酰赖氨酸。对RA患者表达acpa的B细胞的B细胞受体分析表明，acpa发生了广泛的体细胞超突变，这可以促进表位向多个瓜氨酸化表位扩散。鉴于ACPA表位扩散与疾病进展的关联，当引入与其他ptm的交叉反应性时，获得更多的见解是相关的。此外，交叉反应性是否也存在于无RA的acpa阳性受试者中，或局限于将要或已经发生RA的受试者中，也将有助于更好地理解抗修饰蛋白抗体（AMPA）反应的演变。因此，我们分析了疾病前样本和无RA的ACPA阳性个体的ACPA反应的交叉反应性。为此，使用在线补充材料中描述的修饰肽测量不同队列中的ACPA、anti-CarPA和AAPA。首先，我们分析了19名瑞典受试者的AMPA-IgG反应，这些受试者后来发展为RA。正如预期的那样，ACPA可以在疾病发病前数年检测到，抗体水平随着时间的推移而上升（图1A）。我们在抗carpa和AAPA中发现了类似的模式。有趣的是，对于大多数可检测到ACPA、抗carpa和/或AAPA的患者，这些抗体可以在同一时间点检测到，表明它们在发病前几年同时出现。接下来，我们分析了来自基于社区的长崎岛研究中acpa阳性和acpa阴性的日本无RA个体样本中的AMPA水平（图1B，在线补充图S1）。有趣的是，观察到不同个体ampa反应性水平之间存在很强的相关性，表明抗体存在交叉反应性（图1C）。为了实验证实交叉反应性，我们从AMPA值较高的ACPA阳性非ra个体中选择6个样本，分离ACPA、抗carpa和AAPA，并测定分离抗体对三种不同PTMs的反应性。分离的acpa对同质酰化和乙酰化抗原具有高度反应性，反之亦然，这表明非RA个体的AMPA对不同的PTMs也具有交叉反应性（图1D）。这些结果在翻译后修饰纤维蛋白原和FCS上得到了证实（在线补充图S2）。有趣的是，与乙酰化抗原相比，用瓜氨酸化抗原或高瓜氨酸化抗原分离AMPA对瓜氨酸化/高瓜氨酸化肽的反应性更高。这表明ACPA…
Figure 1 (A) ACPA, anti-CarP and AAPA IgG levels, using the CCP4, CHcitP4 and CAcetylP4 peptides as antigen, in arbitrary units per ml (aU/mL) over time of 19 patients with RA before disease onset. Left graphs show the data in years before onset. The heatmap on the right shows a summary of the AMPA IgG levels per time point.(B) ACPA, anti-CarP and AAPA IgG levels in aU/ml of Japanese ACPA-non-RA samples (n= 197), Japanese AcpA+ non RA samples (n= 54), Dutch healthy donors (n= 30) and established patients with RA (n= 29).(C) Correlations of ACPA, anti-CarP and AAPA levels in aU/mL in Japanese ACPA+ non RA samples. R= correlation coefficient.(D) ACPA, anti-CarP and AAPA IgG levels in aU/mL of six samples from Japanese AcpA+ non RA samples after antibody isolation using CCP4, CHcitP4 or CAcetylP4 peptides. ACPA, anticitrullinated protein antibody; AAPA, antiacetylated protein antibody; anti-CarPA, anti-carbamylated protein antibodies; RA, rheumatoid arthritis. acetyllysine, as shown at both the monoclonal and polyclonal antibody level. 2 3 B cell receptor analysis of ACPA-expressing B cells from patients with RA has shown that ACPAs have undergone extensive somatic hypermutation and that this can facilitate epitope spreading to multiple citrullinated epitopes. 4 Given the association of ACPA epitope spreading with progression to disease, it is relevant to obtain more insights when cross-reactivity to other PTMs is introduced. Furthermore, insights in whether cross-reactivity is also present in ACPA-positive subjects without RA or confined to subjects that will—or have developed RA will also help to better understand the evolution of anti-modified protein antibody (AMPA) responses. Therefore, we analysed cross-reactivity of the ACPA response in pre-disease samples and ACPA-positive individuals without RA. To this end, ACPA, anti-CarPA and AAPA in different cohorts were measured using modified peptides as described in online supplemental materials. First, we analysed the AMPA-IgG response in samples from 19 different Swedish subjects who later developed RA. As expected, ACPA could be detected years before disease onset with a rise in antibody level over time (figure 1A). We detected a similar pattern for anti-CarPA and AAPA. Interestingly, for most patients with detectable ACPA, anti-CarPA and/or AAPA, these antibodies could be detected at the same timepoint, indicating their simultaneous appearance years before disease onset. Next, we analysed AMPA levels in samples from ACPA-positive and ACPA-negative Japanese individuals without RA, derived from the community-based Nagasaki Island study (figure 1B, online supplemental figure S1). 5 Intriguingly, a strong correlation between levels of the different individual AMPA-reactivities was observed, pointing to cross-reactivity of the antibodies (figure 1C). To experimentally confirm cross-reactivity, we selected six samples from ACPA-positive non-RA individuals with high AMPA values, isolated ACPA, anti-CarPA and AAPA and determined the reactivity of the isolated antibodies to the three different PTMs. Isolated ACPAs were highly reactive to the homocitrullinated and acetylated antigen and vice versa, showing that AMPA in individuals without RA are also crossreactive towards different PTMs (figure 1D). These results were confirmed on post-translationally modified fibrinogen and FCS (online supplemental figure S2). Interestingly, the reactivity to citrullinated/homocitrullinated peptides was higher when AMPA were isolated with a citrullinated or homocitrullinated antigen than with an acetylated antigen. This suggests cross-reactivity between ACPA …
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DOI：
--
发表时间：
2010
期刊：
--
影响因子：
--
作者：
D. Aletaha;T. Neogi;A. Silman;Julia Funovits;D. Felson;C. Bingham;N. Birnbaum;G. Burmester;V. Byker
通讯作者：
D. Aletaha;T. Neogi;A. Silman;Julia Funovits;D. Felson;C. Bingham;N. Birnbaum;G. Burmester;V. Byker
影响因子：
27.4
作者：
Nordberg, Lena Bugge;Lillegraven, Siri;Haavardsholm, Espen Andre
通讯作者：
Haavardsholm, Espen Andre
影响因子：
27.4
作者：
Kissel, T.;Reijm, S.;Toes, R. E. M.
通讯作者：
Toes, R. E. M.
尽管突变负荷有限，但IgM抗修饰蛋白抗体的交叉反应性。
影响因子：
4.9
作者：
Reijm S;Kissel T;Stoeken-Rijsbergen G;Slot LM;Wortel CM;van Dooren HJ;Levarht NEW;Kampstra ASB;Derksen VFAM;Heer PO;Bang H;Drijfhout JW;Trouw LA;Huizinga TWJ;Rispens T;Scherer HU;Toes REM
通讯作者：
Toes REM
影响因子：
27.4
作者：
Raza, Karim;Stack, Rebecca;Buckley, Christopher D.
通讯作者：
Buckley, Christopher D.