Identification of host factors affecting Chicken Infectious Anemia Virus infection and pathogenicity through RNA-seq and LC-MS

Chicken Infectious Anemia Virus (CIAV) infection causes severe anemia, hematopoietic disorders, and immunosuppression in poultry. However, the mechanisms underlying its pathogenicity remain poorly understood. In this study, RNA sequencing (RNA-seq) was employed to investigate the transcriptional response of MSB1 cells to CIAV infection at 12, 24, and 48 hours post-infection. The results revealed differential gene expression associated with immune response, inflammatory response, apoptosis, and metabolic pathways. Several immune-related factors, including IL18, TRAF1, MYD88, IRF2, and CD28, were found to be downregulated. VP1, the capsid protein of CIAV, plays a significant role in viral pathogenicity. To explore how VP1 affects CIAV replication and pathogenicity, co-immunoprecipitation (Co-IP) was used to identify host proteins interacting with VP1. Liquid chromatography-mass spectrometry (LC-MS) analysis revealed that VP1 interacts with a range of host proteins involved in metabolic pathways, the PI3K-AKT signaling pathway, and ribosomal functions. Based on RNA-seq and LC-MS findings, ANXA6 and HSP90aa1 were identified as key interacting partners, which co-localize in the cytoplasm of infected cells. Both proteins are involved in immune responses and metabolic regulation and were found to be downregulated in CIAV-infected cells. Notably, overexpression of ANXA6 or HSP90aa1 inhibited CIAV infection during the early stages of the viral lifecycle. These findings enhance our understanding of the host factors influencing CIAV infection and provide new insights into potential therapeutic targets for managing CIAV-induced diseases.