Amir Kalvanagh Parisa, Ebtekara Masoumeh, Kokhaei Parviz, Soleimanjahi Hoorieh
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Immunology, Semnan University of Medical Sciences, Semnan, Iran.
Iran J Pharm Res. 2019 Winter;18(1):156-167.
During the 15 years since the discovery of type III human interferons [IFN-λ1(IL-29), IFN-λ2(IL-28A), and IFN-λ3(IL-28B)], numerous biological properties such as anticancer, antiviral, and immunomodulatory activities of this new IFN family have been investigated. Several studies have shown that the encapsulation of pcDNA with PLGA nanoparticles (NPs) protects them against DNase enzyme action and increases the efficiency of gene delivery to the cells. The purpose of this study was to encapsulate pcDNA encoding IFN-λ1 (pIFN-λ1) with a simple and cost-effective method using PLGA NPs. The pIFN-λ1-loaded PLGA NPs were produced by a double-emulsion-solvent evaporation method and characterized in terms of size, size distribution, and zeta potential by DLS and morphologically by SEM and TEM. The bioactivity of NPs was also examined by fluorescent microscopy. The results showed that IFN-λ1 expressed by HEK293T cells could protect HepC-2 cells from the cytopathic effects of EMCV. The NPs were spherical in shape with a mean diameter of 380 ± 3 nm, a zeta potential of -3.3 ± 7.6 mV, an encapsulation efficiency of 75 ± 5%, and a loading capacity of 0.83 ± 0.06. The NPs were also bioactive and easily engulfed by RAW264.7 cells. The pIFN-λ1 could be sustainably released from NPs. Due to the facility and affordability of encapsulation of pIFN-λ1 in the PLGA NPs proposed in this study and the advantages of encapsulation by PLGA, it appeared rational to use pIFN-λ1-loaded NPs instead of naked pIFN-λ1 to determine other unexplained activities of this new cytokine or to use it as an alternative or adjunct to current IFN-α therapy.
自发现III型人类干扰素[IFN-λ1(IL-29)、IFN-λ2(IL-28A)和IFN-λ3(IL-28B)]以来的15年里，人们对这个新的干扰素家族的多种生物学特性，如抗癌、抗病毒和免疫调节活性进行了研究。多项研究表明，用聚乳酸-羟基乙酸共聚物纳米颗粒（NPs）包裹质粒DNA（pcDNA）可保护其免受脱氧核糖核酸酶的作用，并提高基因传递到细胞的效率。本研究的目的是使用聚乳酸-羟基乙酸共聚物纳米颗粒，通过一种简单且经济高效的方法包裹编码IFN-λ1的质粒DNA（pIFN-λ1）。负载pIFN-λ1的聚乳酸-羟基乙酸共聚物纳米颗粒通过双乳液溶剂蒸发法制备，并通过动态光散射（DLS）对其大小、大小分布和zeta电位进行表征，通过扫描电子显微镜（SEM）和透射电子显微镜（TEM）对其形态进行表征。还通过荧光显微镜检查了纳米颗粒的生物活性。结果表明，HEK293T细胞表达的IFN-λ1可保护HepC-2细胞免受脑心肌炎病毒（EMCV）的细胞病变效应。纳米颗粒呈球形，平均直径为380±3nm，zeta电位为-3.3±7.6mV，包封率为75±5%，载药量为0.83±0.06。纳米颗粒也具有生物活性，并且容易被RAW264.7细胞吞噬。pIFN-λ1可以从纳米颗粒中持续释放。由于本研究中提出的将pIFN-λ1包裹在聚乳酸-羟基乙酸共聚物纳米颗粒中的简便性和经济性，以及聚乳酸-羟基乙酸共聚物包裹的优势，使用负载pIFN-λ1的纳米颗粒而非裸pIFN-λ1来确定这种新细胞因子的其他未解释的活性，或将其用作当前IFN-α治疗的替代或辅助手段似乎是合理的。
