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

基础医学与临床 ›› 2010, Vol. 30 ›› Issue (12): 1318-1324.

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

黄丽¹,郭显蓉²,屠宴会³,宋德禄¹,杨浩²,尹洪超²

1. 中国医学科学院基础医学研究所北京协和医学院基础学院2. 中国医学科学院&北京协和医学院基础所病理系3. 中国医学科学院中国协和医科大学药用植物研究所

收稿日期:2010-03-30 修回日期:2010-05-09 出版日期:2010-12-05 发布日期:2010-12-05
通讯作者: 尹洪超

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

HUANG Li ,GUO Xian-rong ,TU Yan-hui ,SONG De-lu ,YANG Hao ,YIN Hong-chao

Institute of Basic Medical Sciences, CAMS & PUMC

Received:2010-03-30 Revised:2010-05-09 Online:2010-12-05 Published:2010-12-05
Contact: YIN Hong-chao

摘要/Abstract

摘要： 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SAa2,3Gal 和SAa2,6Gal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SAa2,3Gal，但SAa2,6Gal含量却很少。Hep-2细胞表面SAa2,3Gal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-a2,3-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。

关键词: H5N1病毒, 呼吸道上皮细胞, 唾液酸-a2, 3-半乳糖, 唾液酸-a2, 6-半乳糖, 细胞死亡

Abstract: Objective To study the attachment and subsequent entry of H5N1 virus to A549 cells and Hep-2 cells. Methods Expression of SAa2,3Gal and SAa2,6Gal on A549 cells and Hep-2 cells was examined by using lectin fluorescence and flow cytometry. H5N1 virus entry into A549 and Hep-2 cells was measured by indirect immuno?uorescence assay. The entry efficiency was determined by Western blot analysis. Results SAa2,3Gal was prevalent in A549 cells and Hep-2 cells, while SAa2,6 Gal was little found. Furthermore, SAa2,3Gal 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 differed between the two cell lines tested. 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 SAa2,3Gal on the cells tested corresponded with the attachment of the H5N1 virus. However, sialic acid only may not sufficient for entry into cells.

黄丽;郭显蓉;屠宴会;宋德禄;杨浩;尹洪超. 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[J]. 基础医学与临床, 2010, 30(12): 1318-1324.

HUANG Li ;GUO Xian-rong ;TU Yan-hui ;SONG De-lu ;YANG Hao ;YIN Hong-chao. Binding of avian influenza A (H5N1) to gangliosides on the surface of A549 cells and Hep-2 cells[J]. Basic & Clinical Medicine, 2010, 30(12): 1318-1324.