Sajjan Umadevi
Centre for Inflammation and Lung Research, Lewis Katz Medical School, Temple University, Philadelphia, PA, USA.
Department of Microbiology, Immunology and Inflammation, Lewis Katz Medical School, Temple University, Philadelphia, PA, USA.
Methods Mol Biol. 2025;2903:21-30. doi: 10.1007/978-1-0716-4410-2_3.
Rhinovirus (RV)-C, discovered in 2006, binds to human cadherin-related family member 3 expressed on ciliated cells. RV-C is the cause of severe respiratory illness in children with asthma. Unlike rhinovirus-A and B, very little is known about the RV-C biology or pathogenic mechanisms. This is because RV-C cannot be propagated using the conventional method that is described for RV-A and RV-B in Chapter 2 . H1HeLa cells or primary lung fibroblasts do not express the receptor for RV-C and, therefore, they are resistant to RV-C infection. Recently, human mucociliary-differentiated immortalized airway epithelial cells expressing cadherin-related family member 3 cells were shown to be suitable for propagating RV-C from clinical samples, but this method may not be sustainable due to the costs of culturing the cells for large-scale virus production. Another method to propagate the virus is to transfect the viral genome into H1HeLa or primary lung fibroblasts to bypass the initial infection step, which is the binding and endocytosis of the virus. RV-C virus propagated by this method was demonstrated to yield relatively high-titer infectious virus suitable for in vitro studies. Here, we describe a method to propagate and purify the RV-C suitable for in vitro and in vivo studies.
鼻病毒C（RV-C）于2006年被发现，它与纤毛细胞上表达的人类钙黏蛋白相关家族成员3结合。RV-C是哮喘患儿严重呼吸道疾病的病因。与鼻病毒A和B不同，人们对RV-C的生物学特性或致病机制知之甚少。这是因为无法使用第2章中描述的用于RV-A和RV-B的常规方法来培养RV-C。H1HeLa细胞或原代肺成纤维细胞不表达RV-C的受体，因此它们对RV-C感染具有抗性。最近，已证明表达钙黏蛋白相关家族成员3的人黏液纤毛分化永生化气道上皮细胞适合从临床样本中培养RV-C，但由于大规模病毒生产所需的细胞培养成本，这种方法可能无法持续。另一种培养该病毒的方法是将病毒基因组转染到H1HeLa细胞或原代肺成纤维细胞中，以绕过病毒的初始感染步骤，即病毒的结合和内吞作用。通过这种方法培养的RV-C病毒被证明能产生滴度相对较高的适合体外研究的感染性病毒。在此，我们描述一种适合体外和体内研究的培养和纯化RV-C的方法。
