GenÇ Rükan, YakuboĞullari Nilgün, Nalbantsoy Ayşe, ÇÖven Fethiye, Bedİr Erdal
Department of Chemical Engineering, Faculty of Engineering, Mersin Turkey.
Department of Bioengineering, Faculty of Engineering, İzmir Institute of Technology, İzmir Turkey.
Turk J Biol. 2020 Oct 13;44(5):304-314. doi: 10.3906/biy-2003-49. eCollection 2020.
Adjuvants are substances that increase the immune response to a given antigen. In the development of novel vaccine adjuvants/systems, saponins are one of the most attractive molecules due to their altered immunomodulatory activities. In this study, we tried to develop PEG (polyethylene glycol)/cholesterol-based lipid nanoparticles (LNPs) to deliver the Astragaloside VII (AST-VII) and potentiate adjuvant properties of AST-VII for the influenza vaccine. In the formation of PEG/cholesterol/AST-VII-based LNPs (PEG300: Chol-AST-VII LNPs), 3 different primary solvents (acetone, ethanol, and chloroform) were evaluated, employing their effects on hydrodynamic particle size, distribution, surface chemistry, and colloidal stability. Prepared nanoparticles were simply admixtured with inactivated influenza antigen (H3N2) and applied to PMA (phorbol 12-myristate 13-acetate)-ionomycin treated human whole blood to evaluate their cytokine release profile. PEG300: Chol-AST-VII LNPs (80.2 ± 7.7 nm) were obtained using chloroform as a desolvation agent. Co-treatment of PMA-ionomycin with AST-VII and PEG300: Chol-AST-VII LNPs significantly increased the levels of IL-2 and IFN-g, compared to PMA-ionomycin alone. In the presence of H3N2, AST-VII was able to augment IL-17A, while PEG300: Chol-AST-VII LNPs stimulated the production of IFN-g. Hemolysis was only observed in PEG300: Chol-AST-VII LNPs (250 μg/mL) treatment. AST-VII and AST-VII-integrated LNPs could be used as efficacious adjuvants for an inactivated H3N2 vaccine in vitro, and cytokine response through Th1/Th17 route was reported.
佐剂是能够增强对特定抗原免疫反应的物质。在新型疫苗佐剂/系统的研发中，皂苷因其免疫调节活性的改变而成为最具吸引力的分子之一。在本研究中，我们试图开发基于聚乙二醇（PEG）/胆固醇的脂质纳米颗粒（LNP）来递送黄芪甲苷VII（AST-VII），并增强AST-VII对流感疫苗的佐剂特性。在制备基于PEG/胆固醇/AST-VII的LNP（PEG300: Chol-AST-VII LNP）时，评估了3种不同的主要溶剂（丙酮、乙醇和氯仿）对流体动力学粒径、分布、表面化学性质和胶体稳定性的影响。将制备的纳米颗粒简单地与灭活流感抗原（H3N2）混合，并应用于经佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）-离子霉素处理的人全血，以评估其细胞因子释放谱。使用氯仿作为去溶剂化剂获得了PEG300: Chol-AST-VII LNP（80.2±7.7 nm）。与单独使用PMA-离子霉素相比，PMA-离子霉素与AST-VII和PEG300: Chol-AST-VII LNP共同处理显著提高了白细胞介素-2（IL-2）和γ干扰素（IFN-γ）的水平。在存在H3N2的情况下，AST-VII能够增强白细胞介素-17A（IL-17A）的产生，而PEG300: Chol-AST-VII LNP刺激IFN-γ的产生。仅在PEG300: Chol-AST-VII LNP（250μg/mL）处理中观察到溶血现象。AST-VII和整合了AST-VII的LNP可作为体外灭活H3N2疫苗的有效佐剂，并报道了通过Th1/Th17途径的细胞因子反应。
