蓝舌病病毒感染对绵羊微血管内皮细胞中Ⅰ型干扰素应答的影响

doi:10.11843/j.issn.0366-6964.2024.12.026

Abstract

Abstract:

Bluetongue virus (BTV) is an arbovirus that seriously harms ruminants such as Sheep, while sheep lung microvascular endothelial cells (SLMECs) constitute a semi-selective barrier in order to study the molecular mechanism of interferon antiviral immune response in BTV infection and SLMECs. In this study, BTV plaque assay was used to induce SLMECs at 18 h, 24 h and 36 h with multiple infection (MOI) 1. The mRNA expression characteristics of interferon-related pathway genes were analyzed by real-time fluorescence quantitative PCR, and the protein expressions of MDA5, TRAF3, RIG-I and TBK1 were analyzed by Western blot. The results showed that the changes of interferon signaling pathway gene expression were most obvious, and the mRNA expressions of RIG-I, MDA5, IKKε, IFN-β, TBK1 and TRAF6 genes were up-regulated, and the differences were extremely significant (P < 0.01) at 36 h of BTV induction, while the mRNA expressions of IFN-α, VISA and USP18 genes were significantly downregulated (P < 0.01). At the protein level, the protein expressions of MDA5, TBK1, RIG-I and TRAF3 were mainly upregulated. This study found that BTV infection can induce an IFN immune response of SLMECs, which lays a foundation for further analysis of the mechanism of IFN-Ⅰ signaling pathway activation in the innate immune response against BTV infection.

Key words: bluetongue virus, type Ⅰ interferon, sheep lung microvascular endothelial cells

CLC Number:

S852.659.4

LUO Shimei, CHEN Yunyi, LI Qisha, ZHOU Yanmei, HU Xuerou, LI Mingyue, WEI Ziyu, LI Fang, MA Xianping, YI Huashan. Effect of Bluetongue Virus Infection on Type Ⅰ Interferon Responses in Sheep Lung Microvascular Endothelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5631-5640.

Figures/Tables 6

Table 1

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目的基因 Target genes	引物序列(5′→3′) Primer sequences(5′→3′)	用途 Usage
β-Actin	F: CCACTGTCGAGTCGCGTCC	qRT-PCR
	R: ATTCCCACCATCACACCCTGG
RIG-Ⅰ	F: CTGACTGCCTCGGTTGGTGTTG	qRT-PCR
	R: CTTGCTCCAGTTCCTCCAGATTGTG
MDA5	F: AGCTTCACCTGGTGTTGGAG	qRT-PCR
	R: TGGCAAACTTCTTGCATGGC
VISA	F: AGCCATCCCAAGGAATCCAG	qRT-PCR
	R: GAGACAGATGGAGACACAGGC
TBK1	F: TGGAAGCGGCAGAGTTAGGTGAA	qRT-PCR
	R: TTCGGATGAGTGCCTTCTTGATGTG
IKKε	F: TGGGAATTCATGCAGAGCACAGCCAATT	qRT-PCR
	R: TGTGGTACCTCAGACATCAGGAGGTGCT
TRAF3	F: CTGCAGGTTTCCTTGTTGCAG	qRT-PCR
	R: TGCTTGGCTGTCTATCACTCG
TRAF6	F: GAGATCCATGACCAGAACTGTCCTT	qRT-PCR
	R: GGCGTGCCAAGTGATTCCTCT
USP18	F: ATCACGAATGAGCAAGGCGT	qRT-PCR
	R: CAACCAGGCCATGAGGGTAG
IFN-α	F: CTCATACACCAGGTCACGCT	qRT-PCR
	R: AGTGTAAAGGTGCACATGACG
IFN-β	F: TAGGCGACACTGTTCGTGTT	qRT-PCR
	R：CCATTCAATTGCCACAGGAGC
VP7	F: ATGGACACTATCGCTGCAAG	RT-PCR
	R：CTACACATAGGCGGCGCGT

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Effect of Bluetongue Virus Infection on Type Ⅰ Interferon Responses in Sheep Lung Microvascular Endothelial Cells

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