Hinz Alicja, Stankiewicz Sylwia, Litewka Jacek Jakub, Ferdek Paweł E, Sochalska Maja, Bzowska Monika
Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.
Doctoral School of Exact and Natural Sciences Jagiellonian University, Kraków, Poland.
Int J Nanomedicine. 2024 Dec 7;19:13165-13181. doi: 10.2147/IJN.S479710. eCollection 2024.
Given the increased use of polyethylene glycol (PEG) in refining the therapeutic activity of medicines, our research focuses on explaining the potential mechanism of immune reactions associated with this polymer. We aim to investigate the interaction of different types of PEG with mouse and human immune cells, thereby contributing to understanding PEG interactions with the immune system and verifying the proinflammatory activity of the tested polymers.
Mouse macrophage and neutrophil cell lines, human peripheral blood mononuclear cells, and polymorphonuclear cells isolated from healthy donors were exposed to various PEGs. ROS, NO, and cytokine production were analyzed using fluorescence intensity, absorbance, or cytometric measurements. Toll-like receptor (TLR) signaling was verified using HEK-blue-reporter cell lines. Finally, neutrophil trap formation was studied using immunofluorescence labeling, and calcium imaging was performed using a Ca-sensitive indicator and fluorescence microscope.
Our findings show that specific PEG and mPEG are not toxic to tested mouse and human cells. However, they exert proinflammatory activity against human immune cells, as evidenced by the increased secretion of proinflammatory cytokines, such as IFN-a2, IFN-γ, TNF-α, MCP-1, IL-8, IL-17A, and IL-23. This phenomenon is independent of PEG signaling via TLR. Additionally, mPEG10 induced the formation of neutrophil extracellular traps and intracellular calcium signaling.
Our finding suggests that some PEG types have proinflammatory activity against human immune cells, manifesting in the upregulated production of cytokines and neutrophils trap releasing.
鉴于聚乙二醇（PEG）在改善药物治疗活性方面的应用日益增加，我们的研究重点是解释与这种聚合物相关的免疫反应的潜在机制。我们旨在研究不同类型的PEG与小鼠和人类免疫细胞的相互作用，从而有助于理解PEG与免疫系统的相互作用，并验证受试聚合物的促炎活性。
将小鼠巨噬细胞和中性粒细胞系、人类外周血单核细胞以及从健康供体分离的多形核细胞暴露于各种PEG中。使用荧光强度、吸光度或细胞计数测量分析活性氧（ROS）、一氧化氮（NO）和细胞因子的产生。使用HEK-蓝色报告细胞系验证Toll样受体（TLR）信号传导。最后，使用免疫荧光标记研究中性粒细胞胞外陷阱的形成，并使用钙敏感指示剂和荧光显微镜进行钙成像。
我们的研究结果表明，特定的PEG和甲氧基聚乙二醇（mPEG）对受试小鼠和人类细胞无毒。然而，它们对人类免疫细胞具有促炎活性，促炎细胞因子如IFN-α2、IFN-γ、TNF-α、MCP-1、IL-8、IL-17A和IL-23的分泌增加证明了这一点。这种现象与通过TLR的PEG信号传导无关。此外，mPEG10诱导了中性粒细胞胞外陷阱的形成和细胞内钙信号传导。
我们的研究结果表明，某些类型的PEG对人类免疫细胞具有促炎活性，表现为细胞因子产生上调和中性粒细胞陷阱释放。
