Allergy Asthma Immunol Res.  2018 Mar;10(2):144-154. 10.4168/aair.2018.10.2.144.

Innate Immune Response to Viral Infections in Primary Bronchial Epithelial Cells is Modified by the Atopic Status of Asthmatic Patients

• Moskwa S
• Piotrowski W
• Marczak J
• PaweÅ‚czyk M
• Lewandowska-Polak A
• JarzÄ™bska M
• Brauncajs
• GÅ‚obiÅ„ska A
• Górski P
• Papadopoulos NG
• Edwards MR
• Johnston S
• Kowalski M

• KMID: 2402974
• DOI: http://doi.org/10.4168/aair.2018.10.2.144

Abstract

PURPOSE
In order to gain an insight into determinants of reported variability in immune responses to respiratory viruses in human bronchial epithelial cells (HBECs) from asthmatics, the responses of HBEC to viral infections were evaluated in HBECs from phenotypically heterogeneous groups of asthmatics and in healthy controls.
METHODS
HBECs were obtained during bronchoscopy from 10 patients with asthma (6 atopic and 4 non-atopic) and from healthy controls (n=9) and grown as undifferentiated cultures. HBECs were infected with parainfluenza virus (PIV)-3 (MOI 0.1) and rhinovirus (RV)-1B (MOI 0.1), or treated with medium alone. The cell supernatants were harvested at 8, 24, and 48 hours. IFN-α, CXCL10 (IP-10), and RANTES (CCL5) were analyzed by using Cytometric Bead Array (CBA), and interferon (IFN)-β and IFN-λ1 by ELISA. Gene expression of IFNs, chemokines, and IFN-regulatory factors (IRF-3 and IRF-7) was determined by using quantitative PCR.
RESULTS
PIV3 and RV1B infections increased IFN-λ1 mRNA expression in HBECs from asthmatics and healthy controls to a similar extent, and virus-induced IFN-λ1 expression correlated positively with IRF-7 expression. Following PIV3 infection, IP-10 protein release and mRNA expression were significantly higher in asthmatics compared to healthy controls (median 36.03-fold). No differences in the release or expression of RANTES, IFN-λ1 protein and mRNA, or IFN-α and IFN-β mRNA between asthmatics and healthy controls were observed. However, when asthmatics were divided according to their atopic status, HBECs from atopic asthmatics (n=6) generated significantly more IFN-λ1 protein and demonstrated higher IFN-α, IFN-β, and IRF-7 mRNA expressions in response to PIV3 compared to non-atopic asthmatics (n=4) and healthy controls (n=9). In response to RV1B infection, IFN-β mRNA expression was lower (12.39-fold at 24 hours and 19.37-fold at 48 hours) in non-atopic asthmatics compared to atopic asthmatics.
CONCLUSIONS
The immune response of HBECs to virus infections may not be deficient in asthmatics, but seems to be modified by atopic status.

Keyword

Asthma; bronchial epithelial cells; interferon; parainfluenza virus; rhinovirus

MeSH Terms

Asthma
Bronchi*
Bronchoscopy
Chemokine CCL5
Chemokines
Enzyme-Linked Immunosorbent Assay
Epithelial Cells*
Gene Expression
Humans
Immunity, Innate*
Interferons
Paramyxoviridae Infections
Polymerase Chain Reaction
Rhinovirus
RNA, Messenger
Chemokine CCL5
Chemokines
Interferons
RNA, Messenger

Figure

• 

  Fig. 1 PIV3 (A) and RV1B (B) replication in HBECs from asthmatic patients (n=5) and healthy controls (n=5). Horizontal line indicates the median.

• 

  Fig. 2 IFN-λ1 protein concentration in cell supernatants (A) and mRNA expression (B) in HBECs post PIV3 infection, and IFN-λ1 protein concentration in cell supernatants (C), and mRNA expression (D) in HBECs post RV1B infection in asthmatic patients (n=10) and healthy controls (n=9). Significant induction in HBECs by PIV3 and RV1B. *P<0.05, **P<0.01 compared to medium control.

• 

  Fig. 3 IRF7 mRNA expression in HBECs post PIV3 (A) and RV1B (B) infection in asthmatic patients (n=10) and healthy controls (n=9). Significant induction in HBECs by PIV3 and RV1B. *P<0.05, **P<0.01 compared to medium control.

• 

  Fig. 4 IP-10 protein (A) and mRNA expression (B) in HBECs post PIV3 infection and IP-10 protein (C) and mRNA expression (D) in HBECs post RV1B infection in asthmatic patients (n=10) and healthy controls (n=9). Significant induction in HBECs by PIV3 and RV1B. *P<0.05, **P<0.01 compared to medium control.

• 

  Fig. 5 IFN-λ1 protein concentration (A), as well as IFN-λ1 (B), IFN-β mRNA (C), and IRF-7 (D) mRNA expression, in HBECs post PIV3 infection in healthy controls (n=9), patients with atopic asthma (n=6), and patients with non-atopic asthma (n=4). Significant induction in HBECs by PIV3.

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