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腺病毒55型对人肠道细胞感染性的实验研究

Infectivity of Human Adenovirus Type 55 to Human Intestinal Cells

  • 摘要:
      目的  明确腺病毒55型(HAdV-55)对人肠道细胞的感染性。
      方法  体外培养人结直肠腺癌细胞Caco-2,以HAdV-3、7、14和55感染,免疫荧光法检测感染细胞内病毒蛋白的表达,荧光定量PCR方法检测不同时间点细胞内和上清中病毒DNA水平,采用腺病毒敏感细胞株HEp-2感染实验检测Caco-2细胞上清中感染性病毒颗粒水平。
      结果  免疫荧光检测结果显示,感染48 h后,Caco-2细胞内HAdV-55病毒蛋白表达为阳性。HAdV-3、7、14、55在Caco-2细胞内均能持续复制和增殖,感染细胞内和上清中病毒DNA水平随感染时间的增加而升高,并且HAdV-55病毒DNA水平明显高于HAdV-3、7和14型。Caco-2上清中HAdV-55感染性病毒颗粒高于HAdV-3、7和14,差异有统计学意义(P<0.05)。采用低剂量病毒〔1×半数组织培养感染剂量(TCID50)〕感染Caco-2细胞,HAdV-55感染孔细胞病变效应(CPE)比HAdV-3、7和14感染孔细胞显著。
      结论  人呼吸道病毒HAdV-55对肠道细胞易感,感染水平高于其他常见的呼吸道感染腺病毒3、7和14型。

     

    Abstract:
      Objective  To examine the infectivity of human adenovirus type 55 (HAdV-55) in human intestinal cells.
      Methods  Caco-2 cells were cultured in vitro, and infected with HAdV-3, 7, 14 and 55. The expression of viral proteins in infected cells was detected with immunofluorescence method. The intracellular and supernatant viral DNA levels were determined with fluorescent quantitative PCR at different points of time. The level of infectious virus particles in the supernatant of Caco-2 cells was determined with adenovirus sensitive HEp-2 infection assay.
      Results  Immunofluorescence assay showed positive result for the expression of HAdV-55 virus protein in Caco-2 cells 48 h post infection. HAdV-3, 7, 14, and 55 showed sustained replication and proliferation in Caco-2 cells. The level of viral DNA in infected cells and the supernatant increased with the infection time, and the viral DNA level of HAdV-55 was significantly higher than those of HAdV-3, 7 and 14. The infectious virus particles of HAdV-55 in Caco-2 supernatant were more than those of HAdV-3, 7 and 14, showing statistically significant difference (P<0.05). Caco-2 cells were infected with low doses of virus (1×TCID50), and the cytopathic effect (CPE) of HAdV-55 infection wells was more significant than that of HAdV-3, 7 and 14 infection wells.
      Conclusion  This study found that human intestinal cells were susceptible to HAdV-55, and the infection level was higher than that of other common respiratory infections caused by adenovirus types 3, 7 and 14.

     

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