Kim Da Been, Lee Sun Min, Geem Kyoung Rok, Kim Jitae, Kim Eui Ho, Lee Dong Wook
Department of Integrative Food, Bioscience and Biotechnology, Chonnam National University, Gwangju 61186, Korea.
Viral Immunology Laboratory, Institut Pasteur Korea, Seongnam 13488, Korea.
Plants (Basel). 2022 Nov 4;11(21):2984. doi: 10.3390/plants11212984.
Influenza viruses are a major public health threat that causes repetitive outbreaks. In recent years, genotype 4 (G4) reassortant Eurasian avian-like (EA) H1N1 (G4 EA H1N1) has garnered attention as a potential novel pandemic strain. The necessity of developing vaccines against G4 EA H1N1 is growing because of the increasing cases of human infection and the low cross-reactivity of the strain with current immunity. In this study, we produced a G4 EA H1N1-derived neuraminidase (G4NA) as a vaccine candidate in . The expressed G4NA was designed to be accumulated in the endoplasmic reticulum (ER). The M-domain of the human receptor-type tyrosine-protein phosphatase C was incorporated into the expression cassette to enhance the translation of G4NA. In addition, the family 3 cellulose-binding module and small ubiquitin-like modifier sequences were used to enable the cost-effective purification and removal of unnecessary domains after purification, respectively. The G4NA produced in plants displayed high solubility and assembled as a tetramer, which is required for the efficacy of an NA-based vaccine. In a mouse immunization model, the G4NA produced in plants could induce significant humoral immune responses. The plant-produced G4NA also stimulated antigen-specific CD4 T cell activation. These G4NA vaccine-induced immune responses were intensified by the administration of the antigen with a vaccine adjuvant. These results suggest that G4NA produced in plants has great potential as a vaccine candidate against G4 EA H1N1.
流感病毒是一种导致反复爆发的重大公共卫生威胁。近年来，基因型4（G4）重配的欧亚禽源样（EA）H1N1（G4 EA H1N1）作为一种潜在的新型大流行毒株受到关注。由于人类感染病例的增加以及该毒株与现有免疫力的交叉反应性较低，开发针对G4 EA H1N1疫苗的必要性日益凸显。在本研究中，我们在植物中生产了一种源自G4 EA H1N1的神经氨酸酶（G4NA）作为候选疫苗。所表达的G4NA被设计在内质网（ER）中积累。将人受体型酪氨酸蛋白磷酸酶C的M结构域整合到表达盒中以增强G4NA的翻译。此外，分别使用3型纤维素结合模块和小泛素样修饰序列，以实现经济高效的纯化以及纯化后去除不必要的结构域。在植物中产生的G4NA具有高溶解度，并组装成四聚体，这是基于NA的疫苗发挥效力所必需的。在小鼠免疫模型中，植物中产生的G4NA可诱导显著的体液免疫反应。植物产生的G4NA还刺激了抗原特异性CD4 T细胞活化。通过与疫苗佐剂一起施用抗原，这些由G4NA疫苗诱导的免疫反应得到增强。这些结果表明，植物中产生的G4NA作为针对G4 EA H1N1的候选疫苗具有巨大潜力。
