H7N9流感病毒在4种细胞中培养情况的研究

Study on the culture of H7N9 influenza in 4 kinds of cells

目的了解H7N9流感病毒在不同细胞中增殖的感染动力学差异。方法将H7N9流感病毒分别接种于鸡成纤维细胞(DF-1)、狗肾细胞(MDCK)、人支气管上皮细胞(BEAS-2B)、人肺腺癌细胞(A549),观察病毒在各细胞内增殖后细胞病变效应(CPE)、血凝实验滴度及50%组织培养细胞感染剂量。结果 H7N9流感病毒在A549细胞中培养36 h可达到血凝效价最高,同时可见明显CPE,在DF-1及MDCK细胞中培养72 h后血凝效价达到最高;在BEAS-2B细胞中增殖不明显,培养液对豚鼠血无明显凝集,且培养超过96 h尚无明显CPE。DF-1细胞、MDCK细胞、A549细胞对病毒的TCID50实验结果分别为10^(-4.34)、10^(-5.13)、10^(-3.84)。结论本实验使用的H7N9流感病毒不能有效感染BEAS-2B细胞并在其中增殖,但是可以在DF-1、MDCK及A549细胞中有效增殖。增殖能力以MDCK细胞最强,DF-1细胞略弱,A549细胞次之。A549细胞对病毒的吸附能力较强,可快速达到增殖的高峰。

Objective To explore the proliferation difference of H7N9 influenza virus in different cells. Methods H7N9 influenza were inoculated on DF- 1,MDCK,BEAS- 2B and A549 cells to observe the cytopathic effect,HA titer of hemagglutination test,50% tissue culture infective dose. Results H7N9 influenza reached the highest HA titer in A549 cells after culturing about 36 hours and had obvious CPE,and the virus got to the highest HA titer in DF- 1 and MDCK cells after culturing for about 72 hours. The virus had not obviously reproduced in BEAS- 2B cells,nutrient fluid had not agglutinated for blood of guinea pigs and had no obvious CPE. The experimental results of TCID50 were 10^- 4. 34（ DF- 1 cells）,10^- 5. 13（ MDCK cells）,10^- 3. 84（ A549 cells）. Conclusion H7N9 influenza can not reproduce in BEAS- 2B cells but can reproduce in DF- 1,MDCK and A549 cells. The proliferation ability of MDCK cells was most powerful,followed by DF- 1 cells and A549 cells. A549 cells can quickly adsorb H7N9 influenza and get to the peak of proliferation.