Department of Chemical and Biomolecular Engineering, Yonsei University, Yonsei-ro 50, Seoul, 120-749, Republic of Korea.
Department of Pharmacy, Korea University College of Pharmacy, Sejong-ro 2511, Sejong, 30019, Republic of Korea.
J Nanobiotechnology. 2020 Mar 24;18(1):54. doi: 10.1186/s12951-020-00611-8.
Influenza viruses (IVs) have become increasingly resistant to antiviral drugs that target neuraminidase and matrix protein 2 due to gene mutations that alter their drug-binding target protein regions. Consequently, almost all recent IV pandemics have exhibited resistance to commercial antiviral vaccines. To overcome this challenge, an antiviral target is needed that is effective regardless of genetic mutations.
In particular, hemagglutinin (HA), a highly conserved surface protein across many IV strains, could be an effective antiviral target as it mediates binding of IVs with host cell receptors, which is crucial for membrane fusion. HA has 6 disulfide bonds that can easily bind with the surfaces of gold nanoparticles. Herein, we fabricated porous gold nanoparticles (PoGNPs) via a surfactant-free emulsion method that exhibited strong affinity for disulfide bonds due to gold-thiol interactions, and provided extensive surface area for these interactions. A remarkable decrease in viral infectivity was demonstrated by increased cell viability results after exposing MDCK cells to various IV strains (H1N1, H3N2, and H9N2) treated with PoGNP. Most of all, the viability of MDCK cells infected with all IV strains increased to 96.8% after PoGNP treatment of the viruses compared to 33.9% cell viability with non-treated viruses. Intracellular viral RNA quantification by real-time RT-PCR also confirmed that PoGNP successfully inhibited viral membrane fusion by blocking the viral entry process through conformational deformation of HA.
We believe that the technique described herein can be further developed for PoGNP-utilized antiviral protection as well as metal nanoparticle-based therapy to treat viral infection. Additionally, facile detection of IAV can be achieved by developing PoGNP as a multiplatform for detection of the virus.
由于流感病毒 (IVs) 的基因发生突变，改变了其药物结合靶蛋白区域，使其对针对神经氨酸酶和基质蛋白 2 的抗病毒药物的耐药性越来越强。因此，几乎所有最近的 IV 大流行都对商业抗病毒疫苗表现出了耐药性。为了克服这一挑战，需要寻找一种无论基因突变如何都有效的抗病毒靶点。
特别是血凝素 (HA)，一种在许多 IV 株中高度保守的表面蛋白，可能是一种有效的抗病毒靶点，因为它介导 IV 与宿主细胞受体的结合，这对于膜融合至关重要。HA 有 6 个二硫键，可以很容易地与金纳米粒子的表面结合。在此，我们通过无表面活性剂乳液法制造了多孔金纳米粒子 (PoGNPs)，由于金 - 硫醇相互作用，这些纳米粒子对二硫键具有很强的亲和力，并为这些相互作用提供了广泛的表面积。暴露于用 PoGNP 处理的各种 IV 株（H1N1、H3N2 和 H9N2）的 MDCK 细胞后，细胞活力增加的结果表明病毒感染性显著降低。最重要的是，与未处理的病毒相比，在用 PoGNP 处理病毒后，所有 IV 株感染的 MDCK 细胞的活力增加到 96.8%，而细胞活力为 33.9%。通过实时 RT-PCR 对细胞内病毒 RNA 进行定量也证实了 PoGNP 通过阻止病毒进入过程中的 HA 构象变形成功抑制了病毒膜融合。
我们相信，本文所述的技术可以进一步发展为利用 PoGNP 进行抗病毒保护以及基于金属纳米粒子的治疗来治疗病毒感染。此外，通过将 PoGNP 开发为病毒检测的多平台，可以实现对 IAV 的简便检测。
