CAJ | 학술논문

目的探讨不同宿主来源的H1N1亚型流感病毒在A549和BEAS-2B细胞的复制情况.方法用分离自人、禽、猪三种宿主的7株H1N1甲型流感病毒分别接种A549和BEAS-2B细胞,分析病毒感染细胞后不同时段的特点;应用受体类型不同的红细胞进行微量血凝试验,检测流感病毒的受体结合特性;同时检测了A549和BEAS-2B细胞表面的受体分布情况.结果三种宿主来源的H1N1亚型流感病毒感染A549细胞,24 h后CPE十分明显,36 h病毒滴度达到最高值;而感染BEAS-2B细胞后,从24 h-120 h CPE都不是很明显,且所有病毒的病毒滴度都很低.对6株H1N1流感病毒的受体结合特性进行了筛查,发现部分测试病毒具有SA a-2,6Gal受体结合特异性.而A549和BEAS-2B细胞表面均含有SA a-2,3Gal及SA a-2,6Gal受体,且A549细胞表面糖受体含量明显高于BEAS-2B细胞.结论不同宿主来源的H1N1亚型流感病毒对A549细胞都易感并能有效增殖复制,而对具有相似受体特性、上皮组织来源的BEAS-2B细胞不易感,提示支持流感病毒有效感染、复制存在宿主内的调节机制.
목적 탐토불동숙주래원적H1N1아형류감병독재A549화BEAS-2B세포적복제정황.방법 용분리자인、금、저삼충숙주적7주H1N1갑형류감병독분별접충A549화BEAS-2B세포,분석병독감염세포후불동시단적특점;응용수체류형불동적홍세포진행미량혈응시험,검측류감병독적수체결합특성;동시검측료A549화BEAS-2B세포표면적수체분포정황.결과 삼충숙주래원적H1N1아형류감병독감염A549세포,24 h후CPE십분명현,36 h병독적도체도최고치;이감염BEAS-2B세포후,종24 h-120 h CPE도불시흔명현,차소유병독적병독적도도흔저.대6주H1N1류감병독적수체결합특성진행료사사,발현부분측시병독구유SA a-2,6Gal수체결합특이성.이A549화BEAS-2B세포표면균함유SA a-2,3Gal급SA a-2,6Gal수체,차A549세포표면당수체함량명현고우BEAS-2B세포.결론 불동숙주래원적H1N1아형류감병독대A549세포도역감병능유효증식복제,이대구유상사수체특성、상피조직래원적BEAS-2B세포불역감,제시지지류감병독유효감염、복제존재숙주내적조절궤제.
Objective Analyze the proliferation of different host H1N1 subtype influenza viruses in A549 and BEAS-2B cells. Methods Human, avain and swine three hosts of the H1N1 influenza viruses infected AS49 and BEAS-2B cells and analyze the characteristics of different periods after inocubation. Determine the receptor binding specificity of influenza virus by hemagglutination ( HA) test with RBCs with two types of receptor. And the receptors on surfaces of AS49 and BEAS-2B cells were tested by flow cytometry. Results The Cell Pathologic Effect(CPE) is obvious after 24 h inoculation in A549 cells by all the H1N1 influenza viruses, moreover,the peak hemagglutinin (HA) and 50% tissue culture cell infected dose (TCID50 ) titers was observed after 36 h of culturing in A549 cells. Otherwise, the CPE is not typical from 24 h-120 h inoculated by the same viruses and the HA,TCID50 titers were keep low all the periods in the BEAS-2B cell after inoculation. The receptor-binding preference of H1N1 viruses used in the study was screened by HA assay and some were found with 2-6-receptor binding affinity. Both SA a-2, 3Gal and SA a-2, 6Cal receptors were detected on A549 and BEAS-2B, furthermore, receptor density on AS49 cells was significantly higher than that of BEAS-2B cells. Conclusion A549 cells were susceptible to human, avian and swine H1N1 influenza viruses infection and permissively for viral replication. However, BEASE-2B cells with similar receptor pattern and epithelium-derived propriety as A549 cells were unsusceptible to their infection and replication. Possible host factors involved in effective viral infection and replication were needed further study.