Luo Qingping, Ahmed Waqas, Dai Yichen, Mohsin Ali, Hang Haifeng, Zhuang Yingping, Guo Meijin
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Vaccines (Basel). 2021 Oct 3;9(10):1128. doi: 10.3390/vaccines9101128.
Porcine circovirus Type 2 (PCV2) is a primary etiological pathogen of post-weaning multi-systemic wasting syndrome (PMWS). The capsid protein of PCV2 is the crucial immunogenic protein which can induce antibody generation and immune responses. However, there is still a lack of efficient PCV2 vaccines with high immunogenicity. In the current study, we developed a novel engineered PCV2 capsid (∆1-41aa)-pFc fusion protein (PCFP), which comprised a truncated capsid protein of PCV2 and a porcine IgG Fc fragment, fused to the capsid protein of PCV2 at the C-terminus. We found that this novel fusion protein could auto-assemble into virus-like nanoparticles with an estimated mean diameter of 22.6 nm, characterized by transmission electron microscopy. Immunization of BALB/c mice with this fusion protein significantly increased the production levels of anti-PCV2-capsid protein antibody in serum. Besides, the virus-like nanoparticles, PCFP was demonstrated to induce efficient cellular immune responses in mice, as evident by the high specific T cell reactivity to the PCFP fusion protein and the high production of the immune cytokines IFN-γ and IL-10 in an ex vivo re-stimulation system. Collectively, these findings demonstrate that the PCV2 truncated capsid subunit Fc-fusion protein can induce both cellular and humoral immune responses, and it displays great application potential.
猪圆环病毒2型(PCV2)是断奶后多系统消耗综合征(PMWS)的主要病原。PCV2的衣壳蛋白是关键的免疫原性蛋白,可诱导抗体产生和免疫反应。然而,目前仍缺乏具有高免疫原性的高效PCV2疫苗。在本研究中,我们构建了一种新型的工程化PCV2衣壳(∆1-41aa)-pFc融合蛋白(PCFP),它由PCV2的截短衣壳蛋白和猪IgG Fc片段组成,在PCV2衣壳蛋白的C端融合。我们发现这种新型融合蛋白可自组装成平均直径约为22.6 nm的病毒样纳米颗粒,通过透射电子显微镜进行表征。用这种融合蛋白免疫BALB/c小鼠可显著提高血清中抗PCV2衣壳蛋白抗体的产生水平。此外,病毒样纳米颗粒PCFP在小鼠中可诱导有效的细胞免疫反应,这在体外再刺激系统中对PCFP融合蛋白的高特异性T细胞反应性以及免疫细胞因子IFN-γ和IL-10的高产量中得到体现。总体而言,这些发现表明PCV2截短衣壳亚基Fc融合蛋白可诱导细胞免疫和体液免疫反应,具有很大的应用潜力。