CAJ | 학술논문

目的:探讨胶质细胞感染流感病毒后的天然免疫反应,检测流感病毒H1N1和H5N1体外感染小鼠小胶质细胞和星形胶质细胞,是否会诱导胶质细胞趋化因子转录水平的变化及其规律.方法:从新生小鼠大脑皮质分离培养神经胶质细胞,并进一步纯化小胶质细胞和星形胶质细胞,经纯度鉴定后,用感染复数为2的流感病毒H1N1和H5N1进行体外感染,8小时后用免疫荧光检测流感病毒核蛋白(NP)的表达,以确认感染细胞比例.在感染早期(6小时)和感染中期(24小时)分别提取细胞RNA,检测趋化因子转录水平的变化.结果:分离得到小鼠的小胶质细胞和星形胶质细胞,病毒感染后超过95%的细胞可以被感染,感染后的小胶质细胞与星形胶质细胞的CCI-3、CCL-5、CXCL-2、CXCL-9和CXCL-10的转录水平发生不同程度的上调,其中CXCL-10的上调幅度最为明显,禽流感病毒H5N1感染能诱导更强烈的上调反应.结论:流感病毒H1N1和H5N1感染小鼠小胶质细胞与星形胶质细胞,可诱导趋化因子转录水平上调.
목적:탐토효질세포감염류감병독후적천연면역반응,검측류감병독H1N1화H5N1체외감염소서소효질세포화성형효질세포,시부회유도효질세포추화인자전록수평적변화급기규률.방법:종신생소서대뇌피질분리배양신경효질세포,병진일보순화소효질세포화성형효질세포,경순도감정후,용감염복수위2적류감병독H1N1화H5N1진행체외감염,8소시후용면역형광검측류감병독핵단백(NP)적표체,이학인감염세포비례.재감염조기(6소시)화감염중기(24소시)분별제취세포RNA,검측추화인자전록수평적변화.결과:분리득도소서적소효질세포화성형효질세포,병독감염후초과95%적세포가이피감염,감염후적소효질세포여성형효질세포적CCI-3、CCL-5、CXCL-2、CXCL-9화CXCL-10적전록수평발생불동정도적상조,기중CXCL-10적상조폭도최위명현,금류감병독H5N1감염능유도경강렬적상조반응.결론:류감병독H1N1화H5N1감염소서소효질세포여성형효질세포,가유도추화인자전록수평상조.
Objective: To investigate the innate immune response of influenza virus-infected glial cells,the transcription levels in chemokines in mouse microglia and astrocytes were detected which pre-infected by human H1N1 or avian H5N1 influenza viruses.Methods: The glial cells isolated from neonatal mice cerebral cortex were cultured and further microglia and astrocytes were purified.The primary mouse microglia and astrocytes were infected in vitro by H1N1 or H5N1 influenza viruses in a multiplicity of infection (MOI) 2.Eight hours post infection,the influenza virus nucleoprotein (NP) was detected by immunofluorescence to identify the proportion of infected cells.The cellular RNA were extracted at 6 h and 24 h to detect the transcriptional level of chemokines by semi-quantitative RT-PCR.Results: More than 95% of the microgha and astrocytes which isolated from mice were infected.The transcription levels of CCL-3,CCL-5,CXCL-2,CXCL-9 and CXCL-10 from infected microglia and astrocytes were upregulated.Futhermore,the mRNA level of CXCL-10 increased much more.In addition,avian H5N1 influenza virus could induce more stronger upregulation of those chemokines than human H1N1 did.Conclusion: The mouse microglia and astro cytes which are infected by H1N1 influenza virus or H5N1 influenza virus could induce upregulation of transcription level of chemokines.