In Vitro and In Vivo Differences in Murine Third Complement Component (C3) Opsonization and Macrophage/Leukocyte Responses to Antibody-Functionalized Iron Oxide Nanoworms.
In Vitro and In Vivo Differences in Murine Third Complement Component (C3) Opsonization and Macrophage/Leukocyte Responses to Antibody-Functionalized Iron Oxide Nanoworms.
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DOI：
10.3389/fimmu.2017.00151
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发表时间：
2017
影响因子：
7.3
通讯作者：
Simberg D
中科院分区：
文献类型：
--
作者：
Wang G;Griffin JI;Inturi S;Brenneman B;Banda NK;Holers VM;Moghimi SM;Simberg D
平衡纳米药物的表面功能化和低免疫识别是一个重大挑战。补体蛋白第三组分（C3）的调理作用在纳米药物的免疫细胞识别中发挥着重要作用。我们使用葡聚糖包被的超顺磁性氧化铁纳米颗粒（SPIO NWs）研究表面功能化对小鼠血清中C3调理作用和随后的体外巨噬细胞/白细胞识别以及静脉注射到小鼠中的影响。先前，我们发现在小鼠血清中，SPIO NW通过补体凝集素途径被C3调理。交联葡聚糖壳与环氧氯丙烷显着降低C3调理和小鼠腹腔巨噬细胞的摄取。用聚乙二醇（PEG）或PEG-抗体（Ab）（~160个IgG分子/颗粒）进一步官能化的交联的双链体（NW）在体外未显示C3调理作用和腹膜巨噬细胞摄取的增加。在尾静脉注射到小鼠中后，普通交联NW和PEG化交联NW显示出非常低的C3调理作用和小鼠白细胞摄取。然而，Ab修饰的交联纳米线在小鼠中显示出显著的C3调理作用和高水平的补体依赖性白细胞摄取。将缀合的Ab的数量减少至46个IgG分子/颗粒显著降低了C3调理作用和白细胞摄取。使用新鲜小鼠来匹卢定血浆而不是血清显示出与体内C3调理作用更好的相关性。这种差异的原因可能与小鼠血清中补体经典途径的已知不稳定性有关。我们的数据表明，需要在纳米颗粒表面功能化与抗体的微调，以避免过度的补体激活和补体介导的免疫摄取在小鼠中，并提出问题，在体外免疫测定的纳米药物拟模仿体内条件。
Balancing surface functionalization and low immune recognition of nanomedicines is a major challenge. Opsonization with the third component of the complement protein (C3) plays a major role in immune cell recognition of nanomedicines. We used dextran-coated superparamagnetic iron oxide nanoworms (SPIO NWs) to study the effect of surface functionalization on C3 opsonization in mouse serum and subsequent macrophage/leukocyte recognition in vitro as well as on intravenous injection into mice. Previously, we found that in mouse serum, SPIO NWs became opsonized with C3 via complement lectin pathway. Crosslinking the dextran shell with epichlorohydrin significantly decreased C3 opsonization and uptake by mouse peritoneal macrophages. Crosslinked nanoworms (NWs) further functionalized with polyethylene glycol (PEG) or with PEG-antibody (Ab) (~160 IgG molecules/particle) did not show an increase in C3 opsonization and peritoneal macrophage uptake in vitro. Following tail vein injection into mice, plain crosslinked NWs and PEGylated crosslinked NWs showed very low C3 opsonization and mouse leukocyte uptake. However, Ab-decorated crosslinked NWs showed significant C3 opsonization and high level of complement-dependent uptake by leukocytes in mice. Decreasing the number of conjugated Abs to 46 IgG molecules/particle significantly reduced C3 opsonization and leukocyte uptake. Using fresh mouse lepirudin plasma rather than serum showed better correlation with C3 opsonization in vivo. The reason for this difference could be related to the known instability of complement classical pathway in mouse sera. Our data illustrate that fine-tuning in nanoparticle surface functionalization with Abs is required to avoid excessive complement activation and complement-mediated immune uptake in mice, and raise issues with in vitro immunological assays of nanomedicines intended to mimic in vivo conditions.