Involvement of complement activation in the pulmonary vasoactivity of polystyrene nanoparticles in pigs: unique surface properties underlying alternative pathway activation and instant opsonization.
Involvement of complement activation in the pulmonary vasoactivity of polystyrene nanoparticles in pigs: unique surface properties underlying alternative pathway activation and instant opsonization.
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DOI：
10.2147/ijn.s161369
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发表时间：
2018
影响因子：
8
通讯作者：
Szebeni J
中科院分区：
文献类型：
--
作者：
Mészáros T;Kozma GT;Shimizu T;Miyahara K;Turjeman K;Ishida T;Barenholz Y;Urbanics R;Szebeni J
已经提出，许多对纳米药物的超敏反应代表补体（C）激活相关的假变态反应（CARPA），并且猪提供了研究该现象的敏感动物模型。然而，最近的一项研究表明，肺动脉高压，猪CARPA的关键症状，在聚苯乙烯纳米颗粒（PS-NP）诱导反应的情况下，不是由C介导的。表征PS-NP并重新检查CARPA对其猪肺反应性的贡献。分别在人和猪血清中测量200、500和750 nm（直径）PS-NPs及其调理作用的C激活，并与相同NPs在猪体内的血流动力学效应相关。PS-NP的物理化学表征包括大小、微电子电位、低温透射电子显微镜和疏水性分析。通过ELISA测量人血清中的C活化，并通过蛋白质印迹和流式细胞术测量猪血清中PS-NPs的调理作用。在猪CARPA模型中定量PS-NP的肺血管活性。PS-NP是具有强负表面电荷的单分散、高度疏水的球体。在人血清中，它们引起C3 a、Bb和sC 5 b-9的大小依赖性显著升高，但不引起C4d。暴露于猪血清在几分钟内导致C5 b-9和调理素iC 3b沉积在NP上，并且调理素iC 3b片段（C3 dg，C3 d）也出现在血清中。PS-NP在与iC 3b碎片化和调理作用进行相同的时间尺度（分钟）上引起猪的重大血流动力学变化，主要是肺动脉高压。人血清中不同PS-NPs激活的C与猪肺动脉高压之间存在显著相关性。PS-NP具有极端的表面性质，与临床使用的纳米药物无关。它们可以通过旁路途径激活C，从而引起猪血清中NP的瞬时调理作用。因此，PS-NP诱导猪超敏反应的机制可能涉及C激活，而不仅仅是C非依赖性反应。这些数据与纳米颗粒诱导的超敏反应的“双重打击”概念一致，涉及CARPA和C-非依赖性假性过敏。
It has been proposed that many hypersensitivity reactions to nanopharmaceuticals represent complement (C)-activation-related pseudoallergy (CARPA), and that pigs provide a sensitive animal model to study the phenomenon. However, a recent study suggested that pulmonary hypertension, the pivotal symptom of porcine CARPA, is not mediated by C in cases of polystyrene nanoparticle (PS-NP)-induced reactions. To characterize PS-NPs and reexamine the contribution of CARPA to their pulmonary reactivity in pigs. C activation by 200, 500, and 750 nm (diameter) PS-NPs and their opsonization were measured in human and pig sera, respectively, and correlated with hemodynamic effects of the same NPs in pigs in vivo. Physicochemical characterization of PS-NPs included size, ζ-potential, cryo-transmission electron microscopy, and hydrophobicity analyses. C activation in human serum was measured by ELISA and opsonization of PS-NPs in pig serum by Western blot and flow cytometry. Pulmonary vasoactivity of PS-NPs was quantified in the porcine CARPA model. PS-NPs are monodisperse, highly hydrophobic spheres with strong negative surface charge. In human serum, they caused size-dependent, significant rises in C3a, Bb, and sC5b-9, but not C4d. Exposure to pig serum led within minutes to deposition of C5b-9 and opsonic iC3b on the NPs, and opsonic iC3b fragments (C3dg, C3d) also appeared in serum. PS-NPs caused major hemodynamic changes in pigs, primarily pulmonary hypertension, on the same time scale (minutes) as iC3b fragmentation and opsonization proceeded. There was significant correlation between C activation by different PS-NPs in human serum and pulmonary hypertension in pigs. PS-NPs have extreme surface properties with no relevance to clinically used nanomedicines. They can activate C via the alternative pathway, entailing instantaneous opsonization of NPs in pig serum. Therefore, rather than being solely C-independent reactivity, the mechanism of PS-NP-induced hypersensitivity in pigs may involve C activation. These data are consistent with the “double-hit” concept of nanoparticle-induced hypersensitivity reactions involving both CARPA and C-independent pseudoallergy.