牛细小病毒 VP1 通过 RIG-I 样受体通路抑制 IFN-β 产生而逃避免疫。

Immune escape of bovine parvovirus by VP1 inhibiting IFN-β production through the RIG-I-like receptor pathway.

机构信息

Hospital, Northwest Minzu University, Lanzhou, 730030, China.

Life Science and Engineering College, Northwest Minzu University, Lanzhou, 730030, China.

出版信息

Int Microbiol. 2023 Nov;26(4):757-764. doi: 10.1007/s10123-023-00330-8. Epub 2023 Jan 27.

DOI:10.1007/s10123-023-00330-8

PMID:36703013

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9879738/

Abstract

OBJECTIVE

The present study aimed to explore if bovine parvovirus (BPV) impacts beta interferon (IFN-β) production and to reveal further molecular mechanism of BPV immune escape.

METHOD

The pCMV-Myc-BPV-VP1 recombinant plasmid was verified with both double-enzyme digestion and sequence. HEK 293 T cells were transfected with this recombinant protein and then infected with the vesicular stomatitis virus (VSV). Expression levels of IFN-β mRNA were detected using qPCR.

RESULTS

The expression level of BPV VP1 mRNA in the pCMV-Myc-BPV-VP1 group was significantly higher than those of the untreated group (UT) and pCMV-Myc vector group. BPV virus copies in bovine turbinate (BT) cells of the BPV-VP1 group were raised (P < 0.05) with an increment of 5.8 × 10. Expression levels of IFN-β mRNA of the BPV VP1 group in HEK 293 T cells were decreased (P < 0.01). Following treatment of TBK1 and IRF3(5D), IFN-β expression levels in HEK 293 T cells were depressed. Additionally, expression levels of TBK1, IRF3(5D), MDA5, and MAVS were less than those of the flag empty vector, respectively.

CONCLUSION

pCMV-Myc-BPV-VP1 could heighten transcription levels of VP1 protein in BT cells, promote BPV proliferation, and ascend the production of IFN-β. Overexpression of pCMV-Myc-BPV-VP decreased IFN-β mRNA expression in HEK 293 T cells and inhibited IFN-β production induced by TBK1 and IRF3(5D). Furthermore, BPV VP1 obviously declined expression levels of TBK1, IRF3(5D), MDA5, and MAVS in the RIG-I-like receptor (RLR) pathway. Our findings revealed a novel mechanism evolved by BPV VP1 to inhibit type I IFN production and provided a solid scientific basis into the immunosuppression of BPV.

摘要

目的

本研究旨在探讨牛细小病毒（BPV）是否影响β干扰素（IFN-β）的产生，并揭示 BPV 免疫逃逸的进一步分子机制。

方法

用双酶切和测序验证 pCMV-Myc-BPV-VP1 重组质粒。用该重组蛋白转染 HEK 293T 细胞，然后用水疱性口炎病毒（VSV）感染。用 qPCR 检测 IFN-β mRNA 的表达水平。

结果

pCMV-Myc-BPV-VP1 组 BPV VP1 mRNA 的表达水平明显高于未处理组（UT）和 pCMV-Myc 载体组。BPV-VP1 组牛鼻甲（BT）细胞中的 BPV 病毒拷贝数增加（P<0.05），增加了 5.8×10。HEK 293T 细胞中 BPV VP1 组 IFN-β mRNA 的表达水平降低（P<0.01）。用 TBK1 和 IRF3（5D）处理后，HEK 293T 细胞中 IFN-β的表达水平降低。此外，TBK1、IRF3（5D）、MDA5 和 MAVS 的表达水平均低于空载 Flag 载体，分别。

结论

pCMV-Myc-BPV-VP1 可提高 BT 细胞中 VP1 蛋白的转录水平，促进 BPV 增殖，增加 IFN-β的产生。pCMV-Myc-BPV-VP 的过表达降低了 HEK 293T 细胞中 IFN-βmRNA 的表达，并抑制了 TBK1 和 IRF3（5D）诱导的 IFN-β产生。此外，BPV VP1 明显降低了 RIG-I 样受体（RLR）通路中 TBK1、IRF3（5D）、MDA5 和 MAVS 的表达水平。我们的研究结果揭示了 BPV VP1 抑制 I 型 IFN 产生的新机制，并为 BPV 的免疫抑制提供了坚实的科学依据。