The co-application of nectin-1 and PRV gD monoclonal antibodies: An effective approach to block pseudorabies virus invasion.

Pseudorabies virus (PRV), a zoonotic alphaherpesvirus threatening swine industries and human health, exploits glycoprotein D (gD)-nectin-1 interactions for host cell entry. Here, we report a dual-target neutralizing strategy combining a novel gD-specific monoclonal neutralization antibody (3F7) with nectin-1-blocking mAb. In vitro neutralization assays demonstrated that 3F7 alone achieved potent cross-species inhibition of PRV infection in porcine kidney cells (PK15), human neuroblastoma cells (SK-N-SH), and human embryonic kidney cells (HEK 293 T) (IC50: 0.56-5.39 μg/mL). Co-administration with nectin-1-targeting mAb synergistically enhanced neutralization efficacy by 0.9-25.8 %, revealing a cooperative mechanism between viral glycoprotein blockade and host receptor interference. This study presents the initial evidence indicating that antibodies targeting both the pathogen and its host entry factor, nectin-1, significantly enhance the anti-PRV activity. These findings highlight a novel and effective strategy for mitigating zoonotic PRV transmission, especially among high-risk populations such as swine workers. Moreover, the study provides a promising avenue for the development of next-generation biologics against emerging zoonotic herpesviruses, offering significant implications for both veterinary and human public health.