Targeting of Plasminogen Activator Inhibitor 1 Improves Fibrinolytic Therapy for Tetracycline-Induced Pleural Injury in Rabbits
Targeting of Plasminogen Activator Inhibitor 1 Improves Fibrinolytic Therapy for Tetracycline-Induced Pleural Injury in Rabbits
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DOI：
10.1165/rcmb.2014-0168oc
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发表时间：
2015-04-01
影响因子：
6.4
通讯作者：
Komissarov, Andrey A.
中科院分区：
文献类型：
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作者：
Florova, Galina;Azghani, Ali;Komissarov, Andrey A.
内源性活性纤溶酶原激活物抑制剂1 （PAI-1）在体内被单克隆抗体（mab）靶向，将其与蛋白酶的反应定向到底物分支。单克隆抗体被用作四环素致胸膜损伤兔胸膜内纤溶治疗（IPFT）的辅助药物。在24小时评估scuPA IPFT（0.25或0.0625 mg/kg）与0.5 mg/kg小鼠IgG或mab （MA-33H1F7和MA-8H9D4）的结果。在IPFT后0、10、20、40分钟和24小时收集胸膜液（PF），分析纤溶酶原活化（PA）、uPA、纤溶活性、总纤溶酶/纤溶酶原、α -巨球蛋白（α M）、单克隆抗体/IgG抗原、游离活性uPA和α M/uPA复合物的水平。抗pai -1单克隆抗体（Anti-PAI-1 mab），而非小鼠IgG，在scuPA最小有效剂量降低8倍（从0.5降至0.0625 mg/kg）的情况下，改善了IPFT的结果（P < 0.05）。PFs在24小时检测到单克隆抗体和IgG。与相同剂量的scuPA单独或与IgG相比，scuPA和抗pai -1单抗治疗可产生更高的PF - uPA酰胺解活性和PA活性，aM/uPA复合物的形成更快，uPA失活更慢。然而，PAI-1靶向并没有显著影响胸膜内纤溶活性或总纤溶蛋白/纤溶酶原和aM抗原水平。以PAI-1为靶点不会引起出血，并且使无效剂量的scuPA能够改善四环素引起的胸膜损伤的结局。pai -1中和单抗通过增加胸膜内PA活性的持久性来改善IPFT。这些结果提示了一种新的、耐受性良好的IPFT策略，可用于临床开发。
Endogenous active plasminogen activator inhibitor 1 (PAI-1) was targeted in vivo with monoclonal antibodies (mAbs) that redirect its reaction with proteinases to the substrate branch. mAbs were used as an adjunct to prourokinase (single-chain [sc] urokinase [uPA]) intrapleural fibrinolytic therapy (IPFT) of tetracycline-induced pleural injury in rabbits. Outcomes of scuPA IPFT (0.25 or 0.0625 mg/kg) with 0.5 mg/kg of mouse IgG or mAbs (MA-33H1F7 and MA-8H9D4) were assessed at 24 hours. Pleural fluid (PF) was collected at 0, 10, 20, and 40 minutes and 24 hours after IPFT and analyzed for plasminogen activating (PA), uPA, fibrinolytic activities, levels of total plasmin/plasminogen, alpha-macroglobulin (alpha M), mAbs/IgG antigens, free active uPA, and alpha M/uPA complexes. Anti-PAI-1 mAbs, but not mouse IgG, delivered with an eightfold reduction in the minimal effective dose of scuPA (from 0.5 to 0.0625 mg/kg), improved the outcome of IPFT (P < 0.05). mAbs and IgG were detectable in PFs at 24 hours. Compared with identical doses of scuPA alone or with IgG, treatment with scuPA and anti-PAI-1 mAbs generated higher PF uPA amidolytic and PA activities, faster formation of aM/uPA complexes, and slower uPA inactivation. However, PAI-1 targeting did not significantly affect intrapleural fibrinolytic activity or levels of total plasmin/plasminogen and aM antigens. Targeting PAI-1 did not induce bleeding, and rendered otherwise ineffective doses of scuPA able to improve outcomes in tetracycline-induced pleural injury. PAI-1-neutralizing mAbs improved IPFT by increasing the durability of intrapleural PA activity. These results suggest a novel, well-tolerated IPFT strategy that is tractable for clinical development.