TRAF3 deficiency in MDCK cells improved sensitivity to the influenza A virus

Yang Le; Jiayou Zhang; Zheng Gong; Zhegang Zhang; Xuanxuan Nian; Xuedan Li; Daiguan Yu; Ning Ma; Rong Zhou; Guomei Zhang; Bo Liu; Lu Yang; Baiqi Fu; Xiuqin Xu; Xiaoming Yang

doi:10.1016/j.heliyon.2023.e19246

TRAF3 deficiency in MDCK cells improved sensitivity to the influenza A virus

Heliyon. 2023 Aug 21;9(9):e19246. doi: 10.1016/j.heliyon.2023.e19246. eCollection 2023 Sep.

Authors

Yang Le^{1

2}, Jiayou Zhang^{1

2}, Zheng Gong^{1

2}, Zhegang Zhang^{1

2}, Xuanxuan Nian^{1

2}, Xuedan Li^{1

2}, Daiguan Yu^{1

2}, Ning Ma^{1

2}, Rong Zhou^{1

2}, Guomei Zhang^{1

2}, Bo Liu^{1

2}, Lu Yang^{1

2}, Baiqi Fu³, Xiuqin Xu³, Xiaoming Yang^{1

2

4}

Affiliations

¹ National Engineering Technology Research Center for Combined Vaccines, 430207, Wuhan, China.
² Wuhan Institute of Biological Products Co.Ltd., 430207, Wuhan, China.
³ Wuhan Institute of Biotechnology, Wuhan, 430075, China.
⁴ China National Biotech Group Company Limited, 100029, Bejing, China.

Abstract

Tumor necrosis factor receptor-associated factor 3 (TRAF3), an adaptor protein, has significant and varying effects on immunity depending on cell types. The role of TRAF3 in Madin-Darby Canine Kidney Epithelial (MDCK) cell resistance to influenza A virus (IVA) remains elusive. In the present study, CRISPR-Cas9 gene editing technology was used to construct the TRAF3 knockout MDCK cells (MDCK-TRAF3^-/-). Hemagglutination assay, plaque assay, transcriptome, and quantitative real-time PCR were performed after IVA infection. The results showed that after IVA infection, HA titers and virus titers were promoted, interferon I-related pathways were significantly blocked, and transcription of several antiviral-related genes was significantly decreased in MDCK-TRAF3^-/- cells. Thus, our study suggests that TRAF3 gene knockout reduced MDCK cell's resistance to IVA, thereby resulting in a promising way for IVA isolation and vaccine manufacturing.

Keywords: CRISPR-Cas9; Influenza; Madin-Darby canine kidney epithelial (MDCK); Tumor necrosis factor receptor-associated factor 3 (TRAF3).