Method to Generate High-Titer Rhinovirus C Suitable for In Vitro and In Vivo Studies

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.