中山大学学报（医学科学版）2026，Vol.47Issue(2)：259-268,10.DOI:10.11714/jsysu.med.YX20250165
基于NOD样受体热蛋白结构域相关蛋白3炎症小体信号筛选猴痘病毒调控炎症的关键蛋白
Screening of the Monkeypox Virus Key Proteins Regulating Inflammation Based on NOD-like Receptor Family Pyrin Domain Containing 3 Inflammasome Signaling
摘要
Abstract
[Objective]To screen and identify the key proteins of Monkeypox Virus(MPXV)that regulate the NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome.[Methods]Using the genomic sequence of the domestic epidemic strain of MPXV as a template to construct an expression plasmid library containing 82 viral genes.To screen for viral genes that significantly inhibit the production of interleukin-1β(IL-1β),the expression plasmids encoding NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),cysteine protease-1(Caspase-1),and pro-IL-1β-Luc reporter gene were co-transfected into HEK293T cells,along with the viral gene plasmids.The signal of the NLRP3 inflammasome was monitored by measuring luciferase activity in the cell supernatant.The inhibition effects were further confirmed by Western Blot(WB),and the interactions between the viral proteins and the components of the NLRP3 inflammasome were determined by Co-immunoprecipitation(Co-IP).Additionally,site-directed mutagenesis was performed to identify the specific interaction motifs.[Results]An NLRP3 inflammasome signal screening system were successfully established and optimized.Screening the MPXV plasmids using this system revealed that B10R most significantly inhibits IL-1β maturation in a dose-dependent manner(P＜0.000 1).Further characterization demonstrated that B10R interacts directly with pro-IL-1β.Notably,the C-terminal motif mutant of B10R(B10Rmut-C,T96A,Y97A,I98A)lost its ability to interact with pro-IL-1β.[Conclusions]The MPXV B10R protein interacts with pro-IL-1β,thereby inhibiting the IL-1β maturation.The C-terminal motif(T96,Y97,I98)of the B10R protein is crucial for function.This study provides important clues and a foundation for further elucidating the molecular mechanisms by which MPXV regulates the NLRP3 inflammasome.关键词
NOD样受体热蛋白结构域相关蛋白3/炎症小体/猴痘病毒/白细胞介素1β/B10RKey words
NOD-like receptor family pyrin domain containing 3/inflammasome/monkeypox virus/interleukin-1β/B10R分类
医药卫生引用本文复制引用
罗铭兴,裴晨辰,龚李云,郑新雷,钱军,刘全,郑君..基于NOD样受体热蛋白结构域相关蛋白3炎症小体信号筛选猴痘病毒调控炎症的关键蛋白[J].中山大学学报（医学科学版）,2026,47(2):259-268,10.基金项目
国家自然科学基金(32570624) （32570624）
广东省科学院发展专项(2022GDASZH-2022010106,2024GDASZH-2024010101) （2022GDASZH-2022010106,2024GDASZH-2024010101）
