Shukla Sourabh, Dorand R Dixon, Myers Jay T, Woods Sarah E, Gulati Neetu M, Stewart Phoebe L, Commandeur Ulrich, Huang Alex Y, Steinmetz Nicole F
Department of Biomedical Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106, United States.
Department of Case Comprehensive Cancer Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106, United States.
ACS Biomater Sci Eng. 2016 May 9;2(5):829-837. doi: 10.1021/acsbiomaterials.6b00060. Epub 2016 Mar 30.
Multiple administrations of nanoparticle-based formulations are often a clinical requirement for drug delivery and diagnostic imaging applications. Steady pharmacokinetics of nanoparticles is desirable to achieve efficient therapeutic or diagnostic outcomes over such repeat administrations. While clearance through mononuclear phagocytic system is a key determinant of nanoparticle persistence , multiple administrations could potentially result in altered pharmacokinetics by evoking innate or adaptive immune responses. Plant viral nanoparticles (VNPs) represent an emerging class of programmable nanoparticle platform technologies that offer a highly organized proteinaceous architecture and multivalency for delivery of large payloads of drugs and molecular contrast agents. These very structural features also render them susceptible to immune recognition and subsequent accelerated systemic clearance that could potentially affect overall efficiency. While the biodistribution and pharmacokinetics of VNPs have been reported, the biological response following repeat administrations remains an understudied area of investigation. Here, we demonstrate that weekly administration of filamentous plant viruses results in the generation of increasing levels of circulating, carrier-specific IgM and IgG antibodies. Furthermore, PVX specific immunoglobulins from the serum of immunized animals quickly form aggregates when incubated with PVX . Such aggregates of VNP-immune complexes are also observed in the mouse vasculature following repeat injections when imaged in real time using intravital two-photon laser scanning microscopy (2P-LSM). The size of aggregates diminishes at later time points, coinciding with antibody class switching from IgM to IgG. Together, our results highlight the need for careful assessment of (viral) nanoparticle-based platform technologies, especially in studying their performance after repeat administration. We also demonstrate the utility of intravital microscopy to aid in this evaluation.
基于纳米颗粒的制剂多次给药通常是药物递送和诊断成像应用的临床要求。纳米颗粒稳定的药代动力学对于在这种重复给药过程中实现有效的治疗或诊断结果是可取的。虽然通过单核吞噬系统的清除是纳米颗粒持久性的关键决定因素，但多次给药可能会通过引发先天性或适应性免疫反应而导致药代动力学改变。植物病毒纳米颗粒（VNP）代表了一类新兴的可编程纳米颗粒平台技术，其提供高度有序的蛋白质结构和多价性，用于递送大量药物和分子造影剂。这些结构特征也使它们易于受到免疫识别和随后加速的全身清除的影响，这可能会潜在地影响整体效率。虽然已经报道了VNP的生物分布和药代动力学，但重复给药后的生物学反应仍然是一个研究不足的领域。在这里，我们证明每周施用丝状植物病毒会导致循环中载体特异性IgM和IgG抗体水平的增加。此外，来自免疫动物血清的PVX特异性免疫球蛋白在与PVX孵育时会迅速形成聚集体。当使用活体双光子激光扫描显微镜（2P-LSM）实时成像时，在重复注射后的小鼠脉管系统中也观察到这种VNP-免疫复合物聚集体。聚集体的大小在后期时间点减小，这与抗体类别从IgM转换为IgG一致。总之，我们的结果强调了仔细评估基于（病毒）纳米颗粒的平台技术的必要性，特别是在研究它们重复给药后的性能时。我们还证明了活体显微镜在这种评估中的实用性。
