中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2012年
2期
130-132
,共3页
呼吸道合胞病毒,人%受体,病毒%干扰素Ⅰ型
呼吸道閤胞病毒,人%受體,病毒%榦擾素Ⅰ型
호흡도합포병독,인%수체,병독%간우소Ⅰ형
Respiratory syncytial virus,human%Receptors,virus%Interferon type Ⅰ
目的 探讨呼吸道合胞病毒( RSV)感染人肺上皮A549细胞后,Toll样受体3(TLR3)的水平变化及其产生的Ⅰ型干扰素的抗病毒作用.方法 RSV感染体外培养的人肺上皮A549细胞,并给予TLR3特异性抗体处理,分别感染4、8、12、16、24h后收集各组细胞.未感染病毒的细胞作为对照组.RT-PCR法检测TLR3、IFN-α、IFN-β,RSV F蛋白的mRNA表达水平变化.结果 RSV感染A549细胞后,TLR3、IFN-α、IFN-β,RSV F蛋白的mRNA表达量均升高且有时间依赖性,TLR3 mRNA在24h表达量是基础表达量的5倍,IFN-α、IFN-β mRNA在24 h表达量是基础表达量的4倍多,RSVF蛋白的mRNA表达量近1.7倍.TLR3抗体预先处理以抑制TLR3受体,再行RSV感染,IFN-α和IFN-β mRNA表达量虽然升高,但较感染组均有所下降,mRNA表达在12 h后显著降低,且IFN-ββ的mRNA表达量下调更明显.但RSV F基因的mRNA表达在12 h、24 h升高有显著性差异.结论 RSV感染A549细胞后可上调TLR3表达,其活化细胞介导产生的Ⅰ型干扰素起抗病毒作用,在一定程度上可抑制病毒的增殖水平.
目的 探討呼吸道閤胞病毒( RSV)感染人肺上皮A549細胞後,Toll樣受體3(TLR3)的水平變化及其產生的Ⅰ型榦擾素的抗病毒作用.方法 RSV感染體外培養的人肺上皮A549細胞,併給予TLR3特異性抗體處理,分彆感染4、8、12、16、24h後收集各組細胞.未感染病毒的細胞作為對照組.RT-PCR法檢測TLR3、IFN-α、IFN-β,RSV F蛋白的mRNA錶達水平變化.結果 RSV感染A549細胞後,TLR3、IFN-α、IFN-β,RSV F蛋白的mRNA錶達量均升高且有時間依賴性,TLR3 mRNA在24h錶達量是基礎錶達量的5倍,IFN-α、IFN-β mRNA在24 h錶達量是基礎錶達量的4倍多,RSVF蛋白的mRNA錶達量近1.7倍.TLR3抗體預先處理以抑製TLR3受體,再行RSV感染,IFN-α和IFN-β mRNA錶達量雖然升高,但較感染組均有所下降,mRNA錶達在12 h後顯著降低,且IFN-ββ的mRNA錶達量下調更明顯.但RSV F基因的mRNA錶達在12 h、24 h升高有顯著性差異.結論 RSV感染A549細胞後可上調TLR3錶達,其活化細胞介導產生的Ⅰ型榦擾素起抗病毒作用,在一定程度上可抑製病毒的增殖水平.
목적 탐토호흡도합포병독( RSV)감염인폐상피A549세포후,Toll양수체3(TLR3)적수평변화급기산생적Ⅰ형간우소적항병독작용.방법 RSV감염체외배양적인폐상피A549세포,병급여TLR3특이성항체처리,분별감염4、8、12、16、24h후수집각조세포.미감염병독적세포작위대조조.RT-PCR법검측TLR3、IFN-α、IFN-β,RSV F단백적mRNA표체수평변화.결과 RSV감염A549세포후,TLR3、IFN-α、IFN-β,RSV F단백적mRNA표체량균승고차유시간의뢰성,TLR3 mRNA재24h표체량시기출표체량적5배,IFN-α、IFN-β mRNA재24 h표체량시기출표체량적4배다,RSVF단백적mRNA표체량근1.7배.TLR3항체예선처리이억제TLR3수체,재행RSV감염,IFN-α화IFN-β mRNA표체량수연승고,단교감염조균유소하강,mRNA표체재12 h후현저강저,차IFN-ββ적mRNA표체량하조경명현.단RSV F기인적mRNA표체재12 h、24 h승고유현저성차이.결론 RSV감염A549세포후가상조TLR3표체,기활화세포개도산생적Ⅰ형간우소기항병독작용,재일정정도상가억제병독적증식수평.
Objective In order to understand the production mechanism of interferon and provide a scientific basis for preventionand clinical therapy,the expression changes of Toll-like receptor (TLR3) mRNA and the role of TLR3 in human lung epithelial cells ( A549 cells) infected with respiratory syncytial virus (RSV) were investigated in this study.Methods RSV infected A549 cells were treated with or without specific antibodies of TLR3 and collected at the selected timepoints after RSV infection (4,8,12,16 and 24h).The expressions of TLR3,IFN-α,IFN-β and RSV F mRNA were evaluated by RT-PCR.Result It was found that RSV infection could markedly up-regulate the mRNA expression of TLR3,IFN-α,IFN-β and RSV F protein in a time-dependent manner as the 24h mRNA expressions of them were 4 times,3 times,3 times and 0.7 times mor than the basic expression,respectively.Treatment of TLR3 specific antibodies,whereas,significantly down-regulated the activation of TLR3.The mRNA expression of IFN-α and IFN-β also decreased accordingly and that of IFN-β reduced more obviously than IFN-α,but that of RSV F protein rose significantly.Conclusion Above data indicate that RSV infection could induce an apparent increase of antiviral genes of IFN-α and IFN-β by activating TLR3 in human lung epithelial cells and the activated cells mediated Type Ⅰ interferon is antiviral,which suggesting that TLR3 might play an important role in antiviral activity of RSV-infected human lung epithelial cells.
