A humanized monoclonal antibody that inhibits platelet-surface ERp72 reveals a role for ERp72 in thrombosis.
A humanized monoclonal antibody that inhibits platelet-surface ERp72 reveals a role for ERp72 in thrombosis.
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DOI：
10.1111/jth.13878
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发表时间：
2018-03
期刊：
影响因子：
--
通讯作者：
Gibbins JM
中科院分区：
文献类型：
--
作者：
Holbrook LM;Sandhar GK;Sasikumar P;Schenk MP;Stainer AR;Sahli KA;Flora GD;Bicknell AB;Gibbins JM
ERp72是一种巯基异构酶。血小板活化过程中，血小板表面ERp72水平升高。我们制备了一种人源化单克隆抗体，可阻断ERp72酶活性（anti - ERp72）。Anti - ERp72抑制血小板功能反应和血栓形成。在内质网内，巯基异构酶调节进入分泌通路的新折叠蛋白质中二硫键的形成和重排，以确保蛋白质正确折叠。除了它们在细胞内的重要性，巯基异构酶最近被发现存在于许多细胞类型的表面，在那里它们对细胞功能很重要。已知在静息血小板表面存在几种硫醇异构酶，包括PDI、ERp5和ERp57，其水平在血小板激活后升高。抑制这些酶的催化活性导致血小板功能降低和血栓形成。我们之前确定ERp72存在于静息血小板表面，并且在血小板激活时水平增加；然而，其在细胞表面的功能作用尚不清楚。我们旨在探讨ERp72在血小板功能中的作用及其在血栓形成中的作用。利用HuCAL技术，生成了完全人源化的Fc - null抗ERp72抗体。筛选了11种抑制ERp72活性的抗体，并选择了最有效的抑制抗体（抗ERp72）用于血小板功能分析的进一步测试。Anti - ERp72抑制血小板聚集、颗粒分泌、钙动员和整合素活化，揭示了细胞外ERp72在血小板活化调控中的重要作用。与此一致的是，抗ERp72输注到小鼠体内可以防止血栓形成。
ERp72 is a thiol isomerase enzyme. ERp72 levels increase at the platelet surface during platelet activation. We generated a humanized monoclonal antibody which blocks ERp72 enzyme activity (anti‐ERp72). Anti‐ERp72 inhibits platelet functional responses and thrombosis. Within the endoplasmic reticulum, thiol isomerase enzymes modulate the formation and rearrangement of disulfide bonds in newly folded proteins entering the secretory pathway to ensure correct protein folding. In addition to their intracellular importance, thiol isomerases have been recently identified to be present on the surface of a number of cell types where they are important for cell function. Several thiol isomerases are known to be present on the resting platelet surface, including PDI, ERp5 and ERp57, and levels are increased following platelet activation. Inhibition of the catalytic activity of these enzymes results in diminished platelet function and thrombosis. We previously determined that ERp72 is present at the resting platelet surface and levels increase upon platelet activation; however, its functional role on the cell surface was unclear. We aimed to investigate the role of ERp72 in platelet function and its role in thrombosis. Using HuCAL technology, fully humanized Fc‐null anti‐ERp72 antibodies were generated. Eleven antibodies were screened for their ability to inhibit ERp72 activity and the most potent inhibitory antibody (anti‐ERp72) selected for further testing in platelet functional assays. Anti‐ERp72 inhibited platelet aggregation, granule secretion, calcium mobilisation and integrin activation, revealing an important role for extracellular ERp72 in the regulation of platelet activation. Consistent with this, infusion of anti‐ERp72 into mice protected against thrombosis.
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DOI：
10.1083/jcb.200602046
发表时间：
2006-06-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Forster ML;Sivick K;Park YN;Arvan P;Lencer WI;Tsai B
通讯作者：
Tsai B
DOI：
10.1161/atvbaha.115.306130
发表时间：
2015-11-01
影响因子：
8.7
作者：
Bye, Alexander P.;Unsworth, Amanda J.;Gibbins, Jonathan M.
通讯作者：
Gibbins, Jonathan M.
影响因子：
4.1
作者：
Chen, Wei;Shang, Wei Hao;Kamata, Tohru
通讯作者：
Kamata, Tohru
影响因子：
5.4
作者：
Swiatkowska, Maria;Szymanski, Jacek;Cierniewski, Czeslaw S.
通讯作者：
Cierniewski, Czeslaw S.
影响因子：
20.3
作者：
Kim, Kyungho;Hahm, Eunsil;Cho, Jaehyung
通讯作者：
Cho, Jaehyung