A multi-glycoprotein synergized recombinant virus system resolved the research dilemma of Epstein-Barr virus entry and facilitated the development of entry-specific drugs or vaccines.

Epstein-Barr virus (EBV) causes multiple severe diseases, and its entry process is a promising prophylactic and therapeutic target. However, EBV entry is intricate with several unresolved mechanisms, and current research methods for it is limited, insensitive, and inefficient. Therefore, novels methods are urgently needed. Herein, we innovatively reported a recombinant virus system rEBV-VSV realizing complicated multi-glycoprotein synergized EBV entry with sensitive detection, which was especially suitable for accurate study of entry mechanisms and effective evaluation of entry-inhibitive therapies. First, rEBV-VSV efficiently entered host cells via multi-synergetic EBV glycoproteins on its membrane and simulated entry features and cellular tropisms of native EBV, also being sensitively detected via bioluminescence. Second, rEBV-VSV was an effective measuring system to evaluate entry-inhibiting potency of EBV-specific antibodies and population's sera in a rapid and high-throughput manner. Third, via flexible mutations and recombination on rEBV-VSV, it was used to accurately study functions of glycoproteins and key residues during EBV entry to better understand entry mechanisms. Especially, via rEBV-VSV, glycoprotein H (gH) mutation L65A+L69A was found to have completely opposite effects on EBV entry into B and epithelia cells, which was manipulated by gp42, and gp42 also powerfully impacted the efficacy of gH-specific antibody E1D1. Conclusively, rEBV-VSV was a satisfactory system to study EBV entry features, mechanisms, and entry-targeted therapies, which could resolve the research dilemma of EBV entry and facilitate the development of EBV-specific drugs and vaccines.