Propagation of Rhinovirus C in Differentiated Immortalized Human Airway HBEC3-KT Epithelial Cells

• Published in: Viruses Vol. 11; no. 3; p. 216
• Main Authors: Nakauchi, Mina, Nagata, Noriyo, Takayama, Ikuyo, Saito, Shinji, Kubo, Hideyuki, Kaida, Atsushi, Oba, Kunihiro, Odagiri, Takato, Kageyama, Tsutomu
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 04.03.2019; MDPI
• Subjects: air–liquid–interface culture; Cell Count; Cell culture; Cell Differentiation; Cell Line, Transformed; Cilia; Clinical isolates; DNA polymerase; Enterovirus - genetics; Enterovirus - growth & development; Enterovirus - physiology; Epithelial cells; Epithelial Cells - virology; Genomes; Goblet cells; Humans; immortalized cells; Propagation; Respiratory System - cytology; Respiratory tract; Rhinovirus; rhinovirus C; Virus Cultivation; Viruses; VP1 protein; United States > US; immortalized cells; air–liquid–interface culture; rhinovirus C; clinical isolates
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v11030216

Rhinoviruses (RVs) are classified into three species: RV-A, B, and C. Unlike RV-A and -B, RV-C cannot be propagated using standard cell culture systems. In order to isolate RV-Cs from clinical specimens and gain a better understanding of their biological properties and pathogenesis, we established air–liquid-interface (ALI) culture methods using HBEC3-KT and HSAEC1-KT immortalized human airway epithelial cells. HBEC3- and HSAEC1-ALI cultures morphologically resembled pseudostratified epithelia with cilia and goblet cells. Two fully sequenced clinical RV-C isolates, RV-C9 and -C53, were propagated in HBEC3-ALI cultures, and increases in viral RNA ranging from 1.71 log10 to 7.06 log10 copies were observed. However, this propagation did not occur in HSAEC1-ALI cultures. Using the HBEC3-ALI culture system, 11 clinical strains of RV-C were isolated from 23 clinical specimens, and of them, nine were passaged and re-propagated. The 11 clinical isolates were classified as RV-C2, -C6, -C9, -C12, -C18, -C23, -C40, and -C53 types according to their VP1 sequences. Our stable HBEC3-ALI culture system is the first cultivable cell model that supports the growth of multiple RV-C virus types from clinical specimens. Thus, the HBEC3-ALI culture system provides a cheap and easy-to-use alternative to existing cell models for isolating and investigating RV-Cs.