M1 polarization of chicken macrophage HD11 can be activated by duck Tembusu virus via MyD88-NF-κB-mediated signaling pathway

Ningwei Geng; Ji Fu; Zehao Lv; Jing Li; Yuxin Kong; Lei Qu; Zhiyun Guo; Jun Zhao; Liya Zhu; Feng Wang; Cui Zhao; Sidang Liu; Zhiyong Hu; Ning Li

doi:10.1016/j.vetmic.2023.109867

M1 polarization of chicken macrophage HD11 can be activated by duck Tembusu virus via MyD88-NF-κB-mediated signaling pathway

Vet Microbiol. 2023 Oct:285:109867. doi: 10.1016/j.vetmic.2023.109867. Epub 2023 Aug 25.

Authors

Ningwei Geng¹, Ji Fu¹, Zehao Lv¹, Jing Li¹, Yuxin Kong¹, Lei Qu¹, Zhiyun Guo¹, Jun Zhao¹, Liya Zhu², Feng Wang³, Cui Zhao³, Sidang Liu⁴, Zhiyong Hu⁵, Ning Li⁶

Affiliations

¹ College of Animal Science and Technology, Shandong Agricultural University, 61 Daizong Road, Taian City, 271018 Shandong Province, China; Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention, Shandong Agricultural University, 61 Daizong Street, Taian City, 271018 Shandong Province, China; Sino-German Cooperative Research Centre for Zoonosis of Animal Origin Shandong Province, Shandong Agricultural University, 61 Daizong Street, Taian City, 271018 Shandong Province, China.
² Animal Husbandry and Veterinary Service Centre of Linshu, Linyi, 276700 Shandong Province, China.
³ Taian City Research Center of Animal Disease Control and Prevention, 8 Hushan East Road, Taian City, 271000 Shandong Province, China.
⁴ College of Animal Science and Technology, Shandong Agricultural University, 61 Daizong Road, Taian City, 271018 Shandong Province, China; Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention, Shandong Agricultural University, 61 Daizong Street, Taian City, 271018 Shandong Province, China.
⁵ College of Animal Science and Technology, Shandong Agricultural University, 61 Daizong Road, Taian City, 271018 Shandong Province, China; Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention, Shandong Agricultural University, 61 Daizong Street, Taian City, 271018 Shandong Province, China. Electronic address: hzy20040111@126.com.
⁶ College of Animal Science and Technology, Shandong Agricultural University, 61 Daizong Road, Taian City, 271018 Shandong Province, China; Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention, Shandong Agricultural University, 61 Daizong Street, Taian City, 271018 Shandong Province, China; Sino-German Cooperative Research Centre for Zoonosis of Animal Origin Shandong Province, Shandong Agricultural University, 61 Daizong Street, Taian City, 271018 Shandong Province, China. Electronic address: SDTaianlining@126.com.

PMID: 37639898
DOI: 10.1016/j.vetmic.2023.109867

Abstract

Duck Tembusu virus (DTMUV) has caused significant economic losses to the global duck industry since its outbreak in 2010. The macrophages act as the key immune cell, and its polarization in different functional states is very important for host's immune responses and microbial infections. Avian macrophages are the main target cells of DTMUV, its polarization induced by DTMUV and the underlying mechanisms were explored in this study. Through quantitative real-time PCR, nitrite assay, and flow cytometry analysis, we found that DTMUV caused severe inflammatory responses in chicken macrophage line HD11 by reprogramming the expression of M1- and M2-associated genes, leading to the polarization of HD11 macrophage to M1-type. In term of mechanism, transcriptomics was performed to analyze the M1-type polarization triggered by DTMUV, it was found that most differential genes were implicated in biological processes, and DTMUV infection significantly activated innate immune signaling pathways, including cytokine-cytokine receptor interaction, MAPK signaling pathway. Moreover, transcription factors NF-κB and AP1 also be activated after viral infection. However, further validation analysis by inhibitors and siRNAs of NF-κB and AP1 showed that NF-κB molecule was essential for DTMUV-induced M1 polarization in HD11 cell, but not AP1. Additionally, the inhibiting assays targeting MyD88 and TRIF molecules were conducted to determine their effect on NF-κB and M1-associated genes upregulated by DTMUV. The results showed that although the inhibition of both MyD88 and TRIF significantly downregulated the mRNA level of NF-κB, but the expression of M1-associated genes such as CD86 was lower in MyD88 inhibition group than in the other group, indicating that the role of MyD88 in mediating M1 polarization induced by DTMUV was more important. Overall, these results demonstrated that DTMUV infection induces M1-type polarization in chicken macrophage HD11 through MyD88-NF-κB signaling pathways. This finding will lay the foundation for further study the pathogenesis of DTMUV, and provide new insights into the prevention and control of this disease.

Keywords: Avian macrophage; Duck Tembusu virus; Inflammatory cytokines; M1-polarization; MyD88-NF-κB signaling.