自然杀伤细胞对人甲型流感病毒抑制作用研究

Study on the Inhibitory Effect of Natural Killer Cells on Human Influenza A Virus

目的:探究自然杀伤细胞(NK细胞)对人甲型流感(FluA)病毒的抑制作用,以提高人体抗病毒感染能力及探索对流感病毒新的治疗手段。方法:进行FluA病毒毒株毒力测定,稀释FluA病毒液,将正常细胞和病毒原液进行对照。收集NK细胞并鉴定,稀释成2.5×10^(4)/mL、5×10^(4)/mL、1×10^(5)/mL、2×10^(5)/mL、4×10^(5)/mL浓度的细胞悬液,使用流式细胞仪进行分析。细胞病变效应(CPE)观测法观察试验细胞病变状态,噻唑蓝(MTT)法计算细胞保护率,分析CPE观测法及MTT法检测NK细胞对FluA病毒抑制作用。结果:在倒置显微镜下显示病毒攻击细胞后,细胞可出现固缩、相互融合形成葡萄串状或星状,随着NK细胞浓度增加,细胞病变反而下降。细胞保护率随着NK细胞效靶比的升高而升高。结论:CPE观测法及MTT法初步发现NK细胞对FluA病毒有一定的抑制作用。在此基础上,可探索针对流感病毒的新治疗手段。

Objective:To explore the inhibitory effect of natural killer cells(NK cells)on human influenza A(FluA)virus,in order to improve the antiviral infection ability of human body and explore new therapeutic methods for influenza virus.Methods:The virulence of FluA virus strain was determined,the FluA virus venom was diluted,and the normal cells and the virus stock were compared.NK cells were collected and identified,diluted into 2.5×10^(4)/mL,5×10^(4)/mL,1×10^(5)/mL,2×10^(5)/mL,4×10^(5)/mL cell suspensions,and analyzed by flow cytometry.Cytopathic effect(CPE)assay was used to observe the pathological status of the test,and the cell protection rate was calculated by MTT assay.The inhibition effect of NK cells on FluA virus was analyzed by CPE assay and MTT assay.Results:The inverted microscope showed that after the virus attacked the cells,the cells could shrink and fuse with each other to form grape strings or stars.With the increase of NK cell concentration,the cytopathy decreased.The cell protection rate increased with the increase of NK cell effect-target ratio.Conclusion:CPE and MTT methods showed that NK cells had certain inhibitory effect on FluA virus.On this basis,new treatments for influenza virus can be explored.