RSV感染对人支气管上皮细胞分泌IL-8和Eotaxin-1的影响

目的探讨呼吸道合胞病毒（RSV）感染对人支气管上皮细胞（NHBE）分泌白介素（IL）-8、嗜酸细胞趋化因子Eotaxin-1的影响。方法用Hep-2细胞扩增病毒,并进行毒力鉴定。建立并鉴定RSV感染NHBE的体外细胞模型。实验分RSV感染组和正常细胞对照组,感染组以不同滴度分为1000、500、100、50 TCID504个组,酶联免疫吸附（ELISA）法检测12、24、48和72h各组IL-8、Eotaxin-1水平。结果实时荧光定量RT-PCR法鉴定细胞模型建立成功。相同感染时间内（12、24、48和72h）,随着感染滴度增加（50、100、500、1000 TCID50）,各感染组IL-8分泌水平均呈上升趋势,与正常细胞对照组比较差异均有统计学意义（P均〈0.05）。相同感染滴度内（50、100、500、1000 TCID50）,随着感染时间增加（24、48和72h）,各感染组IL-8分泌水平均呈上升趋势,与12h组比较均有统计学差异（P均〈0.05）;相同感染时间内（12、24、48和72h）,随着感染滴度增加（50、100、500、1000 TCID50）,各感染组Eotaxin-1分泌水平较正常细胞对照组差异无统计学意义。50、100、500 TCID50相同感染滴度内,随着感染时间增加（24、48和72h）,各感染组Eotaxin-1分泌水平较12h组差异无统计学意义。结论 RSV感染人支气管上皮细胞可显著上调IL-8分泌水平,进一步引起以中性粒细胞浸润为主的气道炎症,而早期的RSV感染对Eotaxin-1影响不大。

The study of interleukin-8 and eotaxin-1 secretion in human bronchial epithelial cell infected with respiratory syncytial virus in vitro

Objective To observe the secretion of IL-8 and Eotaxin-1 in human bronchial epithelial cell infected by RSV in vitro.Methods Hep-2 cell lines were used to amplify RSV and determine its virulence.The RSV infectious titer was determined by TCID50 technique.A NHBE model of RSV infection in vitro was established and identified.The experiment included RSV infection-NHBE group and normal NHBE control group.RSV-infected NHBE group was divided into 5 group based on RSV titers（1000,500,100,50,10 TCID50）in which each group was further divided according to different incubation hours（12h,24h,48h,72h）.The concentrations of IL-8 and Eotaxin-1 in the culture supernatants were detected by enzyme-linked immunosorbent assay（ELISA）.Data were analyzed statistically.Results The mRNA of RSV in NHBE infected-RSV was detected using real time RT-PCR method,which demonstrated that a model of RSV infection in vitro was established successfully.Compared with normal control group,the concentration of IL-8 in RSV-infected NHBE cells supernatants significantly increased.Within same postinoculation time（12h,24h,48h,72h）,IL-8 level increased obviously with raised RSV titers（P0.05）.This trend was noted within same viral titer（10,50,100,500,1000 TCID50）with increased infectious duration,however,compared with 12h group it showed significant difference（P0.05）.No significant differences in Eotaxin-1 levels were seen between these groups.Conclusion IL-8 secretion induced by RSV infection obviously increased with raised RSV titer and infection duration,and further cause infiltration of neutrophils in the airway inflammation.Nevertheless,early RSV infection has little effect on secretion of Eotaxin-1.