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 |