Abstract
RATIONAL Viral infection has been implicated in exacerbations of COPD and asthma, and appropriate in vitro model is required to evaluate infectious mechanism and the efficacies of new therapy. In this study, we compared the replication of virus, such as human rhinovirus (HRV16), respiratory syncytial virus (RSV A2) and influenza virus (WSN33, H1N1) in 3D culture air-liquid interface human nasal epithelial cells (NEC-3D) and NEC monolayer culture (NEC-M).
Methods: HRV16 (1MOI) and RSVA2 (0.1 MOI) (both from ATCC) and WSN33 (1 MOI) (HPA, UK) were infected to normal NEC-3D (Epithelix Sárl, MuciAir®) and NEC-M (Epithelix), and washed out with PBS after 1hr virus absorption. The cells were incubated further up to 10 days. The virus titre in culture supernatant was measured by serial dilutions CPE assay in Hela cells for HRV16, in Hep2 cells for RSV and MDCK cells for H1N1. The results were shown as Log, TCID50 value/20uL. IL-8 was determined by ELISA.
Results: HRV16 replicated in NEC-M and the TCID50 was 3 at maximum. In contrast, HRV16 replicated well in NEC-3D, and TCID50 reached to 6.3 at Day 6 post-infection. RSV A2 replicated well in both cells with peak TCID50 of 4 in NEC-M and 5.3 in NEC-3D. WSN33 only weakly replicated in NEC-M with peak TCID50 of 2, but it replicated well in NEC-3D with peak TCID50 of 6.8 at Day 5. All virus induced IL-8 production by less than 10 fold in NEC-M and by 400 fold in NEC-3D over baseline.
Conclusions: All respiratory virus used in this study replicated more in NEC-3D than NEC-M. This unique and robust 3D culture cell system (MuciAir®) will be a better in vitro model for evaluation of respiratory virus infection.
- © 2011 ERS
