干扰素α2b和λ1体外抗呼吸道合胞病毒活性研究
In vitro anti-respiratory syncytial virus activity of interferon-α2b and interferon-λ1
管恩蕊 1张骞 1陈爱珺 1王超 1黄益曼 1麻粉莲 1郑丽舒1
作者信息
- 1. 中国疾病预防控制中心病毒病预防控制所传染病溯源预警和智能决策全国重点实验室国家卫生健康委员会医学病毒与病毒病重点实验室,北京 100052
- 折叠
摘要
目的 分析干扰素(interferon,IFN)α2b和λ1在Hep2细胞和人呼吸道上皮细胞(human airway epithelium,HAE)中抗呼吸道合胞病毒(respiratory syncytial virus,RSV)病毒活性.方法 在Hep2细胞中,RSV(MOI=0.01)感染后,加入IFN-α2b或IFN-λ1,48 h后观察细胞病变(cytopathic effect,CPE)、RT-qPCR方法测定病毒载量、免疫荧光法检测RSV F蛋白及结晶紫法检测细胞存活率.在HAE细胞中,用IFN-α2b和IFN-λ1预处理24 h后,RSV(MOI=0.01)感染HAE,RT-qPCR方法测定第1～7天培养上清中的病毒载量,免疫荧光法检测RSVF蛋白及空斑法测定第7天培养上清中的病毒滴度.结果 在Hep2细胞中,IFN-α2b和IFN-λ1处理组的CPE较病毒对照组均有所缓解,且均是高浓度组干扰素的CPE轻于低浓度组.不同浓度的IFN-α2b和IFN-λ1均可以显著降低RSV病毒载量(P＜0.001),且高浓度组病毒载量明显低于低浓度组(P＜0.001).此外,IFN-α2b和IFN-λ1均能够降低RSV感染后F蛋白表达量,同时提高细胞存活率.在HAE细胞中,IFN-α2b和IFN-λ1均能抑制RSV病毒复制,降低病毒滴度(P＜0.001)和降低RSV F蛋白表达量.结论 IFN-α2b和IFN-X1在传代细胞Hep2和HAE中对RSV均有较好抗病毒活性,为干扰素抗呼吸道病毒研究提供了数据参考.
Abstract
Objective To analyze antiviral activity against respiratory syncytial virus(RSV)of interferon(IFN)-α2b and IFN-λ1 on Hep2 cells and human airway epithelial(HAE)cells.Methods IFN-α2b or IFN-λ1 was incubated with Hep2 cells after RSV infection,and 48 hours later,the cytopathic effect was observed,the viral load was determined using real time/reverse transcription quantitative polymerase chain reaction(RT qPCR),RSV F protein expression was detected using immunofluorescence,and cell survival rate was detected using crystal violet.HAE cells were incubated with IFN-α2b or IFN-X1 for 24 hours,and then HAE were challenged with RSV.The viral load in the culture supernatant was determined on days 1-7 using RT qPCR,RSV F protein was determined with immunofluorescence and the viral titers in the culture supernatant was detected on day 7 by plaque assay.Results In Hep2 cells,the CPE of the treatment groups(IFN-α2b and IFN-λ1)was alleviated compared to the virus control group,and the CPE of the high concentration group was lighter than that of the low concentration group.Different concentrations of IFN-α2b and IFN-λ1 could significantly reduce the viral load of RSV(P＜0.001),and the viral load of the high concentration group was significantly lower than that of the low concentration group(P＜0.001).In addition,IFN-α2b and IFN-λ1 could reduce the RSV F protein expression after RSV infection and improve cell survival rate.In HAE cells,IFN-α2b and IFN-λ1 could inhibit RSV virus replication,reduce virus titers(P＜0.001)and reduce RSV F protein expression.Conclusions IFN-α2b and IFN-λ1 both showed great antiviral activity against RSV in Hep2 and HAE cells,providing data reference for the study of interferon against respiratory viruses.
关键词
呼吸道合胞病毒/IFN-α2b/IFN-λ1/人呼吸道上皮细胞Key words
Respiratory syncytial virus/IFN-α2b/IFN-λ1/Human airway epithelium引用本文复制引用
基金项目
国家重点研发计划(2022YFC2602202)
出版年
2024
NETL
NSTL
万方数据