virus-like particles of sars-like coronavirus formed by membrane proteins from different origins demonstrate stimulating activity in human dendritic cells病毒样颗粒的类似非典冠状病毒膜蛋白形成的不同起源的演示人体树突状细胞刺激活动.pdfVIP

Virus-Like Particles of SARS-Like Coronavirus Formed by Membrane Proteins from Different Origins Demonstrate Stimulating Activity in Human Dendritic Cells 1,3,4 1 1 1,2 1 1,2 1,2 1 Bingke Bai , Qinxue Hu , Hui Hu , Peng Zhou , Zhengli Shi , Jin Meng , Baojing Lu , Yi Huang , 3 1 Panyong Mao , Hanzhong Wang * 1 State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People’s Republic of China, 2 Graduate School of the Chinese Academy of Sciences, Beijing, People’s Republic of China, 3 Department of Virology, Institute of Infectious Disease, Beijing, People’s Republic of China, 4 Chinese People’s Liberation Army Postgraduate Medical School, Beijing, People’s Republic of China Abstract The pathogenesis of SARS coronavirus (CoV) remains poorly understood. In the current study, two recombinant baculovirus were generated to express the spike (S) protein of SARS-like coronavirus (SL-CoV) isolated from bats (vAcBS) and the envelope (E) and membrane (M) proteins of SARS-CoV, respectively. Co-infection of insect cells with these two recombinant baculoviruses led to self-assembly of virus-like particles (BVLPs) as demonstrated by electron microscopy. Incorporation of S protein of vAcBS (BS) into VLPs was confirmed by western blot and immunogold labeling. Such BVLPs up-regulated the level of CD40, CD80, CD86, CD83, and enhanced the secretion of IL-6, IL-10 and TNF-a in immature dendritic cells (DCs). Immune responses were compared in immature DCs inoculated with BVLPs or with VLPs formed by S, E and M proteins of human SARS-CoV. BVLPs showed a stronger ability to stimulate DCs in terms of cytokine induction as eviden