# 4D-DIA Proteomic Analysis of IPEC-J2 Cells Infected with Porcine Group A Rotavirus G9P[23] Strain

## TL;DR

This study uses advanced proteomic analysis to explore how porcine rotavirus affects intestinal cells, revealing key proteins and pathways involved in infection and immune response.

## Contribution

The study introduces 4D-DIA proteomic analysis to investigate host-pathogen interactions in porcine rotavirus infection, identifying novel differentially abundant proteins and their biological roles.

## Key findings

- 4D-DIA identified 8725 cellular proteins, with 279 more abundant and 356 under-detected proteins in infected cells.
- Western blot confirmed proteomic data for S100A8, DAPK2, and FTL proteins.
- Bioinformatics revealed DAPs involved in immune response, signal transduction, and metabolic pathways.

## Abstract

Porcine rotavirus (PoRV) is a significant pathogen causing gastroenteritis in pigs and has recently emerged as an epidemic in China. In this study, 4D-DIA proteomic analysis was performed on PoRV-infected IPEC-J2 cells, identifying 8725 cellular proteins, including 279 more abundant and 356 under-detected proteins. Western blot validation confirmed consistent expression patterns of S100A8, DAPK2, and FTL with the proteomic data. Bioinformatics analysis revealed that these DAPs were involved in critical biological processes such as immune response, signal transduction, and metabolic pathways. RT-qPCR analysis demonstrated pronounced inflammatory responses during infection. These findings provide critical insights into pathogenic mechanisms and host defense strategies underlying PoRV infection.

Porcine rotavirus (PoRV) is one of the most devastating enteric pathogens causing gastroenteritis in pigs, particularly the sudden occurrence in recent years in China. To elucidate host–pathogen interactions and molecular mechanisms underlying PoRV pathogenesis, four-dimensional (4D) data-independent acquisition (DIA) proteomic (4D-DIA) analysis was performed to comprehensively quantify the differentially abundant proteins (DAPs) in PoRV-infected IPEC-J2 cells. A total of 8725 cellular proteins were identified with 279 more abundant and 356 down abundant proteins. A Western blot showed that the abundance of SA100A8, DAPK2, and FTL were in accordance with the acquired proteomic data using 4D-DIA analysis. Bioinformatics analyses of GO and KEGG demonstrated that various DAPs are involved in crucial biological processes and signaling pathways, such as immune response, signal transduction, metabolic pathways, autophagy, endoplasmic reticulum (ER) stress, and mitochondrial dysfunction. Notably, inflammatory features of host response upon PoRV infection were highlighted, with RT-qPCR confirming the significant upregulation of IL-1α, IL-6, IL-8, TNF-α, STAT1, and IRF9 transcript levels during infection. Altogether, our preliminary findings advance our understanding of PoRV pathogenesis and may shed light on identifying potential targets for the prevention and control of PoRV-associated gastroenteritis.