Broad neutralization of CSFV with novel monoclonal antibodies in vivo

Classical swine fever is a highly contagious disease in China. Although vaccination against Classical swine fever virus (CSFV) has been widely carried out in China, CSFV cases still emerge in an endless stream. Therefore, it is necessary to take new antiviral measures to eliminate CSFV. Glycoprotein E2 of CSFV is the major vaccine candidate that confers protective immunity. Thus, in this study, a batch of neutralizing monoclonal antibodies (mAbs) against E2, as alternative antiviral strategies, were produced. Among them, mAbs 6D10, 8D8 and 3C12 presented neutralizing reactivity against CSFV in a dose-dependent manner. Based on truncated overlapping fragments of E2 and mutants, three linear neutralizing epitopes were identified highly conserved in various CSFV strains. Epitopes ⁸YRYAIS¹³ and ²⁵⁴HECLIG²⁵⁹ were reported for the first time. All the three epitopes are involved in virus internalization and attachment as shown in pre- or post-attachment neutralization. Recombinant polypeptides carrying epitopes successfully inhibit virus infection in PK-15 cells, indicating epitopes were located in receptor-binding domain (RBD). Further, both prophylactic and therapeutic functions of neutralizing antibody were evaluated in rabbits upon CSFV challenge, confirming the efficacy in vivo. These findings provide alternative antiviral strategies against CSFV and deepen the understanding in E2 function during virus entry.