Selection of receptor-binding variants of human influenza A and B viruses in baby hamster kidney cells.
Selection of receptor-binding variants of human influenza A and B viruses in baby hamster kidney cells.
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DOI：
10.1006/viro.1999.9892
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发表时间：
1999-09-15
期刊：
影响因子：
3.7
通讯作者：
Webster, RG
中科院分区：
文献类型：
--
作者：
Govorkova, EA;Matrosovich, MN;Webster, RG
人流感病毒在鸡胚尿囊腔中的培养导致选择在血凝素（HA）分子的球状头部上具有氨基酸取代的受体结合变体。通过在Madin达比犬肾（MDCK）细胞中培养人病毒可以避免这种选择。在本研究中，我们测试了是否婴儿仓鼠肾（BHK）细胞选择受体结合突变体的人流感病毒。在MDCK细胞中从临床样品中分离出H1N1、H3 N2和B型流感病毒后，我们将它们在BHK细胞或鸡蛋中传代。BHK培养的病毒与MDCK培养的病毒的不同之处在于HA中的突变：225 D -> G（H1N1病毒），128 T-> A和2261 -> V（H3 N2），以及187 N-> D（B型）（H3编号）。通过在卵中传代相同MDCK生长的亲本来选择具有不同取代的变体：141 L-> H、208 R-> H和225 D-> G（H1N1）、194 L-> 1（H3 N2）和137 G-> R（B）。与MDCK培养的病毒相比，BHK和鸡蛋培养的病毒对BHK细胞和鸡胚绒毛尿囊细胞的细胞膜以及含3 '-唾液酸半乳糖的合成唾液酸共聚物具有更高的亲和力。相比之下，突变体对含6 '-唾液酸-（N-乙酰乳糖胺）的唾液酸共聚物的亲和力的变化从阴性到阳性不等。用连接特异性凝集素进行的免疫激活细胞分选分析表明，BHK细胞表面含6 ′-唾液酸[-（N-乙酰乳糖胺）]受体的密度明显低于MDCK细胞，这为人病毒在前者细胞中的生长限制提供了解释。我们的数据表明，培养的BHK细胞，像鸡蛋，可以选择受体结合的变种人流感病毒。(C)北京：科学出版社.
Cultivation of human influenza viruses in the allantoic cavity of embryonated chicken eggs leads to a selection of receptor-binding variants with amino acid substitutions on the globular head of the hemagglutinin (HA) molecule. Such selection can be avoided by growing the human viruses in Madin Darby canine kidney (MDCK) cells. In the present study, we tested whether baby hamster kidney (BHK) cells select receptor-binding mutants of human influenza viruses. After isolating H1N1, H3N2, and type B influenza Viruses from clinical samples in MDCK cells, we passaged them in either BHK cells or chicken eggs. The BHK-grown viruses differed from their MDCK-grown counterparts by virtue of mutations in the HA: 225 D --> G (H1N1 virus), 128T --> A and 2261 --> V (H3N2), and 187N --> D (type B) (H3 numbering). Variants with different substitutions were selected by passaging of the same MDCK-grown parents in eggs: 141L --> H, 208R --> H, and 225D --> G (H1N1), 194L --> l (H3N2), and 137G --> R (B). Compared with their MDCK-grown counterparts, both BHK- and egg-grown viruses possessed a higher affinity for the cellular membranes of BHK cells and of the chorioallantoic cells of chicken embryos and for a 3'-sialylgalactose-containing synthetic sialylglycopolymer. By contrast, changes in the affinity of mutants for a 6'-sialyl-(N-acetyllactosamine)-containing sialylglycopolymer varied from negative to positive. Fluorescence-activated cell-sorting analysis with linkage-specific lectins showed that the density of the 6'-sialy[-(N-acetyllactosamine)-containing receptors is substantially lower on the surface of BHK cells than on MDCK cells, providing an explanation for the growth restriction of human viruses in the former cells. Our data demonstrate that cultures of BHK cells, like eggs, can select receptor-binding variants of human influenza viruses. (C) 1999 Academic Press.