Streptococcal-vimentin cross-reactive antibodies induce microvascular cardiac endothelial proinflammatory phenotype in rheumatic heart disease
Streptococcal-vimentin cross-reactive antibodies induce microvascular cardiac endothelial proinflammatory phenotype in rheumatic heart disease
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DOI：
10.1111/cei.12135
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发表时间：
2013-09-01
影响因子：
4.6
通讯作者：
Alessandri, C.
中科院分区：
文献类型：
--
作者：
Delunardo, F.;Scalzi, V.;Alessandri, C.
风湿性心脏病（RHD）的特征是存在抗A组链球菌抗体和抗内皮细胞抗体（AECA）。链球菌抗原和自身蛋白之间的分子模拟是风湿热发病机制的标志。我们的目的是确定，在风湿性心脏病患者，特异性的自身抗体与链球菌蛋白交叉反应的内皮自身抗原，并评估其在诱导内皮损伤的作用。我们使用了免疫蛋白质组学方法与内皮细胞表面膜蛋白，以确定自身抗原AECA识别140 RHD患者。分析了纯化抗体与链球菌蛋白的交叉反应性。通过比较链球菌抗原与人抗原的氨基酸序列，发现了血清交叉反应抗体识别的同源肽。为了研究白细胞介素（IL）-1R相关激酶（IRAK 1）和核因子-B（NF-B）的活化，我们对来自未刺激或刺激的人微血管心脏内皮细胞（HMVEC-C）的全提取物蛋白进行了Western印迹分析。粘附分子的表达和释放的促炎细胞因子和生长因子进行了研究的多重珠基于免疫试剂盒。我们在49%的RHD AECA阳性患者的血清中观察到抗波形蛋白抗体。纯化的抗波形蛋白抗体与热休克蛋白（HSP）70和链球菌蛋白酶链球菌蛋白的交叉反应性。将链球菌HSP 70和链蛋白酶的氨基酸序列与人波形蛋白进行比较，发现了两个可被血清交叉反应抗体识别的同源肽。这些抗体能够刺激HMVEC-C诱导IRAK和NF-B活化、粘附分子表达以及促炎细胞因子和生长因子的释放。总之，链球菌-波形蛋白交叉反应抗体能够通过放大风湿性心脏病的炎症反应来激活微血管心脏内皮。
Rheumatic heart disease (RHD) is characterized by the presence of anti-streptococcal group A antibodies and anti-endothelial cell antibodies (AECA). Molecular mimicry between streptococcal antigens and self proteins is a hallmark of the pathogenesis of rheumatic fever. We aimed to identify, in RHD patients, autoantibodies specific to endothelial autoantigens cross-reactive with streptococcal proteins and to evaluate their role in inducing endothelial damage. We used an immunoproteomic approach with endothelial cell-surface membrane proteins in order to identify autoantigens recognized by AECA of 140 RHD patients. Cross-reactivity of purified antibodies with streptococcal proteins was analysed. Homologous peptides recognized by serum cross-reactive antibodies were found through comparing the amino acid sequence of streptococcal antigens with human antigens. To investigate interleukin (IL)-1R-associated kinase (IRAK1) and nuclear factor-B (NF-B) activation, we performed a Western blot analysis of whole extracts proteins from unstimulated or stimulated human microvascular cardiac endothelial cells (HMVEC-C). Adhesion molecule expression and release of proinflammatory cytokines and growth factors were studied by multiplex bead based immunoassay kits. We observed anti-vimentin antibodies in sera from 49% RHD AECA-positive patients. Cross-reactivity of purified anti-vimentin antibodies with heat shock protein (HSP)70 and streptopain streptococcal proteins was shown. Comparing the amino acid sequence of streptococcal HSP70 and streptopain with human vimentin, we found two homologous peptides recognized by serum cross-reactive antibodies. These antibodies were able to stimulate HMVEC-C inducing IRAK and NF-B activation, adhesion molecule expression and release of proinflammatory cytokines and growth factors. In conclusion, streptococcal-vimentin cross-reactive antibodies were able to activate microvascular cardiac endothelium by amplifying the inflammatory response in RHD.