VLA-2 (α2β1) integrin promotes rotavirus entry into cells but is not necessary for rotavirus attachment
VLA-2 (α2β1) integrin promotes rotavirus entry into cells but is not necessary for rotavirus attachment
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DOI：
10.1128/jvi.76.3.1109-1123.2002
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发表时间：
2002-02-01
影响因子：
5.4
通讯作者：
Estes, MK
中科院分区：
文献类型：
--
作者：
Ciarlet, M;Crawford, SE;Estes, MK
为了鉴定轮状病毒受体，我们测试了 46 个不同物种和组织来源的细胞系对三种 N-乙酰神经氨酸（唾液酸）（SA）依赖性和五种 SA 独立性轮状病毒株感染的敏感性。对SA依赖性或SA非依赖性轮状病毒感染的易感性根据所测试的细胞系和所使用的感染复数（MOI）而变化。肾或肠来源的细胞以及来自乳腺、胃、骨或肺的转化细胞系都容易受到轮状病毒感染，这表明宿主组织范围比之前认识的更广泛。研究发现，中国仓鼠卵巢 (CHO)、幼仓鼠肾 (BHK-21)、豚鼠结肠 (GPC-16)、大鼠小肠 (Rie1) 和小鼠十二指肠 (MODE-K) 细胞即使在 MOIs 为 100 或 500 时也仅支持有限的轮状病毒复制，但通过脂转染将轮状病毒颗粒递送到细胞质中可实现有效的轮状病毒复制。轮状病毒细胞附着蛋白，即外部衣壳刺突蛋白 VP4，含有由 VLA-2 (alpha2beta1) 整联蛋白识别的序列 GDE(A)，为了测试 VLA-2 是否参与轮状病毒附着和进入，我们测量了缺乏 VLA-2 的 CHO 细胞和用人 alpha2 亚基 (CHOalpha2) 或人 alpha2 和 beta1 亚基转染的 CHO 细胞的感染情况VLA-2 的 (CHOalpha2beta1)。在表达 VLA-2 的 CHO 细胞中，SA 依赖性或 SA 不依赖性轮状病毒株的感染效率比亲代 CHO 细胞高 2 至 10 倍，并且抗 VLA-2 抗体可阻断感染敏感性的增加。然而，轮状病毒与 CHO、CHOα2 和 CHOα2β1 细胞的结合水平是相当的，并且与与易感猴肾 (MA104) 细胞或人结肠腺癌（Caco-2、HT-29 和 T-84）细胞的结合相比没有增加，并且结合不被人 α2 亚基抗体阻断。尽管VLA-2整合素促进CHO细胞中的轮状病毒感染，但显然VLA-2整合素单独并不负责轮状病毒细胞的附着和进入。因此，VLA-2不参与轮状病毒与细胞的初始附着，但可能在附着后水平发挥作用。
In an attempt to identify the rotavirus receptor, we tested 46 cell lines of different species and tissue origins for susceptibility to infection by three N-acetyl-neuraminic (sialic) acid (SA)-dependent and five SA-independent rotavirus strains. Susceptibility to SA-dependent or SA-independent rotavirus infection varied depending on the cell line tested and the multiplicity of infection (MOI) used. Cells of renal or intestinal origin and transformed cell lines derived from breast, stomach, bone, or lung were all susceptible to rotavirus infection, indicating a wider host tissue range than previously appreciated. Chinese hamster ovary (CHO), baby hamster kidney (BHK-21), guinea pig colon (GPC-16), rat small intestine (Rie1), and mouse duodenum (MODE-K) cells were found to support only limited rotavirus replication even at MOIs of 100 or 500, but delivery of rotavirus particles into the cytoplasm by lipofection resulted in efficient rotavirus replication. The rotavirus cell attachment protein, the outer capsid spike protein VP4, contains the sequence GDE(A) recognized by the VLA-2 (alpha2beta1) integrin, and to test if VLA-2 is involved in rotavirus attachment and entry, we measured infection in CHO cells that lack VLA-2 and CHO cells transfected with the human alpha2 subunit (CHOalpha2) or with both the human alpha2 and beta1 subunits (CHOalpha2beta1) of VLA-2. Infection by SA-dependent or SA-independent rotavirus strains was 2- to 10-fold more productive in VLA-2-expressing CHO cells than in parental CHO cells, and the increased susceptibility to infection was blocked with anti-VLA-2 antibody. However, the levels of binding of rotavirus to CHO, CHOalpha2, and CHOalpha2beta1 cells were equivalent and were not increased over binding to susceptible monkey kidney (MA104) cells or human colonic adenocarcinoma (Caco-2, HT-29, and T-84) cells, and binding was not blocked by antibody to the human alpha2 subunit. Although the VLA-2 integrin promotes rotavirus infection in CHO cells, it is clear that the VLA-2 integrin alone is not responsible for rotavirus cell attachment and entry. Therefore, VLA-2 is not involved in the initial attachment of rotavirus to cells but may play a role at a postattachment level.