Title Propagation of Rhinovirus C in Differentiated Immortalized Human Airway HBEC3-KT Epithelial Cells.
Author Nakauchi, Mina; Nagata, Noriyo; Takayama, Ikuyo; Saito, Shinji; Kubo, Hideyuki; Kaida, Atsushi; Oba, Kunihiro; Odagiri, Takato; Kageyama, Tsutomu
Journal Viruses Publication Year/Month 2019-Mar
PMID 30836639 PMCID PMC6466094
Affiliation + expend 1.Influenza Virus Research Center, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan. nakauchi@nih.go.jp.

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.

StrainID RV Species Serotype Length(nt) Country Year Strain Name
ANC04239 A 28 873 Japan 2016 0088-OsakaC-JPN-2016
ANC04244 A 10 864 Japan 2015 0441-OsakaC-JPN-2015
ANC04249 A 58 870 Japan 2015 0030-OsakaC-JPN-2015
ANC04254 A 10 865 Japan 2016 28-SGH-JPN-2016
ANC04259 A 46 879 Japan 2015 29-SGH-JPN-2015
ANC04264 A 82 868 Japan 2014 54-SGH-JPN-2014
ANM04240 A 59 858 Japan 2016 0087-OsakaC-JPN-2016
ANM04245 A 54 858 Japan 2015 0058-OsakaC-JPN-2015
ANM04250 A 58 870 Japan 2015 0025-OsakaC-JPN-2015
ANM04255 A 12 861 Japan 2016 25-SGH-JPN-2016
ANM04260 A 16 855 Japan 2015 24-SGH-JPN-2015
ANM04265 A 24 855 Japan 2014 48-SGH-JPN-2014
ANP04237 A 45 881 Japan 2016 0157-OsakaC-JPN-2016
ANP04242 A 40 858 Japan 2016 0071-OsakaC-JPN-2016
ANP04247 A 49 849 Japan 2015 0044-OsakaC-JPN-2015
ANP04252 A 58 870 Japan 2015 0008-OsakaC-JPN-2015
ANP04257 A 19 870 Japan 2015 54-SGH-JPN-2015
ANP04262 A 88 870 Japan 2015 12-SGH-JPN-2015
ANS04238 A 75 868 Japan 2016 0123-OsakaC-JPN-2016
ANS04243 A 21 861 Japan 2016 0063-OsakaC-JPN-2016
ANS04248 A 16 855 Japan 2015 0035-OsakaC-JPN-2015
ANS04253 A 18 861 Japan 2016 59-SGH-JPN-2016
ANS04258 A 101 867 Japan 2015 41-SGH-JPN-2015
ANS04263 A 24 855 Japan 2015 9-SGH-JPN-2015
ANZ04241 A 81 855 Japan 2016 0073-OsakaC-JPN-2016
ANZ04246 A 34 861 Japan 2015 0049-OsakaC-JPN-2015
ANZ04251 A 82 867 Japan 2015 0019-OsakaC-JPN-2015
ANZ04256 A 18 861 Japan 2016 4-SGH-JPN-2016
ANZ04261 A 88 870 Japan 2015 21-SGH-JPN-2015
CNC02869 C 9 816 Japan 2015 47-SGH-JPN-2015
CNC02874 C 23 825 Japan 2016 0105-OsakaC-JPN-2016
CNM02865 C 18 7033 Japan 2014 C18-NIID JPN-2014
CNM02870 C 53 822 Japan 4-SGH-JPN-2015
CNM02875 C 2 813 Japan 0095-OsakaC-JPN-2016
CNP02867 C 9 6935 Japan 2015 C9-NIID JPN-2015
CNP02872 C 18 843 Japan 57-SGH-JPN-2014
CNP02877 C 6 825 Japan 0064-OsakaC-JPN-2016
CNS02868 C 40 813 Japan 63-SGH-JPN-2016
CNS02873 C 12 834 Japan 0153-OsakaC-JPN-2016
CNS02878 C 6 825 Japan 0060-OsakaC-JPN-2016
CNZ02866 C 53 7033 Japan 2015 C53-NIID JPN-2015
CNZ02871 C 12 834 Japan 59-SGH-JPN-2014
CNZ02876 C 6 825 Japan 0089-OsakaC-JPN-2016
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