A549细胞和Hep-2细胞表面糖链类型鉴定及其与H5N1型禽流感病毒结合的特性

Binding of avian influenza A (H5N1) to gangliosides on the surface of A549 cells and Hep-2 cells

目的研究H5N1病毒对A549细胞和Hep-2细胞的黏附和进入特性。方法用凝集素染色技术和流式细胞术检测A549细胞和Hep-2细胞表面SAα2,3Gal和SAα2,6Gal的表达;用间接免疫荧光法检测H5N1病毒进入细胞情况;用Western blot法分析病毒进入A549细胞和Hep-2细胞的效率。结果 A549细胞和Hep-2细胞表面有大量SAα2,3Gal,但SAα2,6Gal含量却很少。Hep-2细胞表面SAα2,3Gal受体的表达水平高于A549细胞。H5N1病毒能够进入A549细胞和Hep-2细胞,而且H5N1病毒对A549细胞的亲嗜性更强。与A549细胞相比,Hep-2细胞对H5N1病毒诱导的细胞死亡更敏感。结论细胞表面唾液酸-α2,3-半乳糖的表达与H5N1病毒的黏附一致。然而,唾液酸受体可能不是介导病毒进入的唯一因素。

Objective To study the attachment and subsequent entry of H5N1 virus into A549 cells and Hep-2 cells.Methods Expression of SAα2,3Gal and SAα2,6Gal on A549 cells and Hep-2 cells was examined by lectin fluorescence and flow cytometry.H5N1 virus in A549 and Hep-2 cells were measured by indirect immunofluorescence assay.The entry efficiency was determined by Western blot analysis.Results SAα2,3Gal was prevalent in A549 cells and Hep-2 cells,while SAα2,6 Gal was little found.Furthermore,SAα2,3Gal expression was more regularly observed in Hep-2 cells rather than A549 cells.The H5N1 virus tested could enter A549 cells and Hep-2 cells.However,viral entry efficiency was different between the two cell lines.A549 cells were found to be more susceptible to avian influenza than Hep-2 cells.H5N1-induced cell death was inefficient in A549 cells than Hep-2 cells.Conclusion The expression of SAα2,3Gal on the cells tested corresponded with the attachment of the H5N1 virus.However,sialic acid only may not be sufficient for entry into cells.