Ex vivo assays to detect complement activation in complementopathies.
Ex vivo assays to detect complement activation in complementopathies.
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DOI：
10.1016/j.clim.2020.108616
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发表时间：
2020-12
期刊：
影响因子：
--
通讯作者：
Brodsky RA
中科院分区：
文献类型：
--
作者：
Yuan X;Yu J;Gerber G;Chaturvedi S;Cole M;Chen H;Metjian A;Sperati CJ;Braunstein EM;Brodsky RA
在补体驱动的血栓性微血管病中，补体激活调节失败导致终末器官损伤。改进的Ham （mHam）测试测量补体介导的体外有核细胞杀伤，但缺乏验证性分析和可靠的阳性对照。我们证明了C5b-9在TF1 PIGAnull细胞表面的积累与mHam中的细胞杀伤有关。我们还发现，唾液酸酶处理细胞或在人血清中添加志贺毒素1比眼镜蛇毒液因子或脂多糖更可靠地作为mHam的阳性对照。在添加补体途径特异性抑制剂（抗c5抗体或因子D抑制剂ACH-145951）的情况下，同时进行mHam和测量C5b-9在GVB++或GVB0 MgEGTA缓冲液中的积累，可用于定位补体调节缺陷。随着更多靶向补体抑制剂的出现，这些检测可能有助于选择补体介导疾病患者的个性化治疗方法。
In complement-driven thrombotic microangiopathies, failure to regulate complement activation leads to end-organ damage. The modified Ham (mHam) test measures complement-mediated killing of a nucleated cell in vitro but lacks a confirmatory assay and reliable positive controls. We demonstrate that C5b-9 accumulation on the surface of TF1 PIGAnull cells correlates with cell killing in the mHam. We also show that Sialidase treatment of cells or addition of Shiga toxin 1 to human serum serve as a more reliable positive control for the mHam than cobra venom factor or lipopolysaccharide. Simultaneously performing the mHam and measuring C5b-9 accumulation either in GVB++ or GVB0 MgEGTA buffer with the addition of complement pathway specific inhibitors (anti-C5 antibody or a factor D inhibitor, ACH-145951) can be used to localize defects in complement regulation. As more targeted complement inhibitors become available, these assays may aid in the selection of personalized treatments for patients with complement-mediated diseases.
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DOI：
10.4049/jimmunol.1501919
发表时间：
2016-01-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Harder MJ;Anliker M;Höchsmann B;Simmet T;Huber-Lang M;Schrezenmeier H;Ricklin D;Lambris JD;Barlow PN;Schmidt CQ
通讯作者：
Schmidt CQ
影响因子：
20.3
作者：
Chaturvedi, Shruti;Braunstein, Evan M.;Brodsky, Robert A.
通讯作者：
Brodsky, Robert A.
影响因子：
4.4
作者：
Orth, Dorothea;Khan, Abdul Basit;Wuerzner, Reinhard
通讯作者：
Wuerzner, Reinhard
影响因子：
7.5
作者：
Rotz, Seth J.;Luebbering, Nathan;Davies, Stella M.
通讯作者：
Davies, Stella M.
影响因子：
2.6
作者：
Savage, William J.;Barber, James P.;Brodsky, Robert A.
通讯作者：
Brodsky, Robert A.