Identification of Major Histocompatibility Complex Class I C Molecule as an Attachment Factor That Facilitates Coronavirus HKU1 Spike-Mediated Infection
Identification of Major Histocompatibility Complex Class I C Molecule as an Attachment Factor That Facilitates Coronavirus HKU1 Spike-Mediated Infection
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DOI：
10.1128/jvi.01387-08
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发表时间：
2009-01-15
影响因子：
5.4
通讯作者：
Yuen, Kwok-Yung
中科院分区：
文献类型：
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作者：
Chan, Che Man;Lau, Susanna K. P.;Yuen, Kwok-Yung
人类冠状病毒HKU 1（HCoV-HKU 1）是最近发现的与全球呼吸道感染相关的人类冠状病毒。在这项研究中，我们已经确定了主要组织相容性复合体I类C分子（HLA-C）作为促进HCoV-HKU 1刺突（S）介导的感染的附着因子。使用人免疫缺陷病毒1型衍生的报告病毒组装HCoV-HKU 1 S假型病毒，所述报告病毒携带HCoV-HKU 1的人密码子优化的刺突。我们鉴定了人肺泡上皮A549细胞为测试的那些中对HCoV-HKU 1 S假型感染最敏感的细胞系。显示A549细胞结合纯化的可溶性HCoV-HKU 1 S1-600糖肽。为了寻找HCoV-HKU 1的功能性受体，构建了A549 cDNA表达文库，并将其转导到非允许的幼仓鼠肾细胞系BHK-21中。分选结合具有C-末端FLAG的可溶性HCoV-HKU 1 S1-600糖蛋白的转导细胞。两个独立克隆的测序显示编码HLA-C的cDNA插入片段。通过RNAi沉默和抗HLA-C抗体抑制HLA-C表达或功能使A549细胞的HCoV-HKU 1 S假型病毒感染降低62%至65%，而用神经氨酸酶预处理细胞仅使这种感染降低13%。当HLA-C在另一个非允许细胞系NIH-3 T3中组成型表达时，定量PCR显示HCoV-HKU 1 S假型病毒与细胞表面的结合增加了200倍，但细胞对HCoV-HKU 1 S假型病毒感染仍然不敏感。我们的数据表明HLA-C参与HCoV-HKU 1与A549细胞的附着，并且是促进细胞进入的潜在候选者。然而，A549细胞上的其他未知表面蛋白可能在病毒进入期间被HCoV-HKU 1的S糖蛋白伴随利用。需要进一步的研究来阐明HCoV-HKU 1的其他假定受体或辅助受体以及HCoV-HKU 1 S介导的细胞进入机制。
Human coronavirus HKU1 (HCoV-HKU1) is a recently discovered human coronavirus associated with respiratory tract infections worldwide. In this study, we have identified the major histocompatibility complex class I C molecule (HLA-C) as an attachment factor in facilitating HCoV-HKU1 spike (S)-mediated infection. HCoV-HKU1 S pseudotyped virus was assembled using a human immunodeficiency virus type 1-derived reporter virus harboring the human codon-optimized spike of HCoV-HKU1. We identified human alveolar epithelial A549 cells as the most susceptible cell line among those tested to infection by HCoV-HKU1 S pseudotypes. A549 cells were shown to bind purified soluble HCoV-HKU1 S1-600 glycopeptide. To search for the functional receptor for HCoV-HKU1, an A549 cDNA expression library was constructed and transduced into the nonpermissive, baby hamster kidney cells line BHK-21. Transduced cells that bind soluble HCoV-HKU1 S1-600 glycoprotein with C-terminal FLAG were sorted. Sequencing of two independent clones revealed cDNA inserts encoding HLA-C. Inhibition of HLA-C expression or function by RNAi silencing and anti-HLA-C antibody decreased HCoV-HKU1 S pseudotyped virus infection of A549 cells by 62 to 65%, whereas pretreatment of cells with neuraminidase decreased such infection by only 13%. When HLA-C was constitutively expressed in another nonpermissive cell line, NIH-3T3, quantitative PCR showed that the binding of HCoV-HKU1 S pseudotyped virus to cell surfaces was increased by 200-fold, but the cells remained nonsusceptible to HCoV-HKU1 S pseudotyped virus infection. Our data suggest that HLA-C is involved in the attachment of HCoV-HKU1 to A549 cells and is a potential candidate to facilitate cell entry. However, other unknown surface proteins on A549 cells may be concomitantly utilized by S glycoprotein of HCoV-HKU1 during viral entry. Further studies are required to elucidate other putative receptors or coreceptors for HCoV-HKU1 and the mechanism of HCoV-HKU1 S-mediated cell entry.