Tao Wenqian, Ziemer Katherine S, Gill Harvinder S
Department of Chemical Engineering, Texas Tech University, Lubbock, TX 79409, USA.
Nanomedicine (Lond). 2014 Feb;9(2):237-51. doi: 10.2217/nnm.13.58. Epub 2013 Jul 5.
This study aimed to develop a novel influenza A vaccine by conjugating the highly conserved extracellular region of the matrix 2 protein (M2e) of influenza A virus to gold nanoparticles (AuNPs) and to test the vaccine in a mouse influenza challenge model.
MATERIALS & METHODS: Citrate-reduced AuNPs (diameter: 12 nm) were synthesized, and characterized by transmission electron microscopy and dynamic light scattering. M2e was conjugated to AuNPs through thiol-gold interactions to form M2e-AuNP conjugates. Particle stability was confirmed by UV-visible spectra, and M2e conjugation was further characterized by x-ray photoelectron spectroscopy. Mice were immunized with M2e-AuNPs with or without CpG (cytosine-guanine rich oligonucleotide) as an adjuvant with appropriate control groups. Sera was collected and M2e-specific immunoglobulin (IgG) was measured, and immunized mice were challenged with PR8-H1N1 influenza virus.
M2e-capped AuNPs could be lyophilized and stably resuspended in water. Intranasal vaccination of mice with M2e-AuNP conjugates induced M2e-specific IgG serum antibodies, which significantly increased upon addition of soluble CpG as adjuvant. Upon challenge with lethal PR8, mice vaccinated with M2e-AuNP conjugates were only partially protected, while mice that received soluble CpG as adjuvant in addition to M2e-AuNP were fully protected.
Overall, this study demonstrates the potential of using the M2e-AuNP conjugates with CpG as an adjuvant as a platform for developing an influenza A vaccine.
本研究旨在通过将甲型流感病毒基质2蛋白(M2e)的高度保守细胞外区域与金纳米颗粒(AuNPs)偶联来开发一种新型甲型流感疫苗,并在小鼠流感攻毒模型中对该疫苗进行测试。
合成了柠檬酸盐还原的AuNPs(直径:12 nm),并通过透射电子显微镜和动态光散射进行表征。M2e通过硫醇-金相互作用与AuNPs偶联,形成M2e-AuNP偶联物。通过紫外可见光谱确认颗粒稳定性,并用X射线光电子能谱进一步表征M2e偶联情况。用含或不含CpG(富含胞嘧啶-鸟嘌呤的寡核苷酸)作为佐剂的M2e-AuNPs免疫小鼠,并设置适当的对照组。收集血清并测量M2e特异性免疫球蛋白(IgG),用PR8-H1N1流感病毒对免疫小鼠进行攻毒。
M2e包被的AuNPs可以冻干并稳定地重新悬浮于水中。用M2e-AuNP偶联物经鼻内接种小鼠可诱导产生M2e特异性IgG血清抗体,添加可溶性CpG作为佐剂后该抗体显著增加。用致死剂量的PR8攻毒时,接种M2e-AuNP偶联物的小鼠仅得到部分保护,而除M2e-AuNP外还接受可溶性CpG作为佐剂的小鼠则得到完全保护。
总体而言,本研究证明了使用M2e-AuNP偶联物与CpG作为佐剂作为开发甲型流感疫苗平台的潜力。