Phenotypic Responses of Differentiated Asthmatic Human Airway Epithelial Cultures to Rhinovirus

Abstract

ObjectivesHuman airway epithelial cells are the principal target of human rhinovirus (HRV), a common cold pathogen that triggers the majority of asthma exacerbations. The objectives of this study were 1) to evaluate an in vitro air liquid interface cultured human airway epithelial cell model for HRV infection, and 2) to identify gene expression patterns associated with asthma intrinsically and/or after HRV infection using this model.MethodsAir-liquid interface (ALI) human airway epithelial cell cultures were prepared from 6 asthmatic and 6 non-asthmatic donors. The effects of rhinovirus RV-A16 on ALI cultures were compared. Genome-wide gene expression changes in ALI cultures following HRV infection at 24 hours post exposure were further analyzed using RNA-seq technology. Cellular gene expression and cytokine/chemokine secretion were further evaluated by qPCR and a Luminex-based protein assay, respectively.Main ResultsALI cultures were readily infected by HRV. RNA-seq analysis of HRV infected ALI cultures identified sets of genes associated with asthma specific viral responses. These genes are related to inflammatory pathways, epithelial structure and remodeling and cilium assembly and function, including those described previously (e.g. CCL5, CXCL10 and CX3CL1, MUC5AC, CDHR3), and novel ones that were identified for the first time in this study (e.g. CCRL1).ConclusionsALI-cultured human airway epithelial cells challenged with HRV are a useful translational model for the study of HRV-induced responses in airway epithelial cells, given that gene expression profile using this model largely recapitulates some important patterns of gene responses in patients during clinical HRV infection. Furthermore, our data emphasize that both abnormal airway epithelial structure and inflammatory signaling are two important asthma signatures, which can be further exacerbated by HRV infection.

Human Rhinovirus Infection of Epithelial Cells Modulates Airway Smooth Muscle Migration

“…CCL5 is the dominant chemoattractant present in HRV-15 CM, although CXCL8 may also contribute. These data, together with earlier reports that HRV infected epithelial cells show expression of genes related to airway remodeling (33), and release a number of growth factors and matrix proteins linked to airway remodeling (34)(35)(36), further support the concept that recurrent HRV infections may contribute to the initiation and progression of airway remodeling in asthma. Future studies will need to determine if the current observations are relevant to cells obtained from asthmatic subjects (both children and adults), while clinical studies of either naturally-acquired or experimental HRV infections will confirm whether these observations occur in vivo.…”

Reduced CDHR3 expression in children wheezing with rhinovirus

“…Recent findings, that epithelial cells from asthmatics cultured ex vivo showed lower mRNA expression of CDHR3 than cells from non‐asthmatics, agree with our findings in peripheral leukocytes in preschool children with wheezing compared with healthy children. Interestingly, they also showed that the expression was further reduced at RV infection with RV A16 and the level correlated with RV infection .…”

Rhinoviruses and Their Receptors: Implications for Allergic Disease

“…CDHR3 subcellular localization has not been mapped yet; however, high level of its expression was found in ciliated cells in differentiated airway epithelium [Griggs et al ., manuscript in preparation]. Recent RNA-seq analysis of RV-A16 infected ALI cultures found that CDHR3 mRNA expression at baseline was slightly lower in asthmatic epithelial cells compared to non-asthmatic cells ( p = 0.02), however, the CDHR3 genotype was not provided for these donors [90]. Interestingly, in contrast to ICAM-1 and LDLR, CDHR3 expression was further decreased upon viral infection and its reduction was highly correlated with viral infection (Pearson correlation p -value = 1.6 × 10 −4 ) indicating different regulatory mechanisms.…”

“…Interestingly, in contrast to ICAM-1 and LDLR, CDHR3 expression was further decreased upon viral infection and its reduction was highly correlated with viral infection (Pearson correlation p -value = 1.6 × 10 −4 ) indicating different regulatory mechanisms. Rhinovirus-induced disruption of the barrier function of airway epithelial cells cultured at ALI along with downregulation of some tight and adherens junction mRNAs and proteins have been also documented [90–92]. …”