α1-Antitrypsin reduces rhinovirus infection in primary human airway epithelial cells exposed to cigarette smoke

Abstract

Human rhinovirus (HRV) infections target airway epithelium and are the leading cause of acute exacerbations of COPD. Cigarette smoke (CS) increases the severity of viral infections, but there is no effective therapy for HRV infection. We determined whether α1-antitrypsin (A1AT) reduces HRV-16 infection in CS-exposed primary human airway epithelial cells. Brushed bronchial epithelial cells from normal subjects and patients diagnosed with COPD were cultured at air–liquid interface to induce mucociliary differentiation. These cells were treated with A1AT or bovine serum albumin for 2 hours and then exposed to air or whole cigarette smoke (WCS) with or without HRV-16 (5×104 50% Tissue Culture Infective Dose [TCID50]/transwell) infection for 24 hours. WCS exposure significantly increased viral load by an average of fivefold and decreased the expression of antiviral genes interferon-λ1, OAS1, and MX1. When A1AT was added to WCS-exposed cells, viral load significantly decreased by an average of 29-fold. HRV-16 infection significantly increased HRV-16 receptor intercellular adhesion molecule-1 messenger RNA expression in air-exposed cells, which was decreased by A1AT. A1AT-mediated reduction of viral load was not accompanied by increased epithelial antiviral gene expression or by inhibiting the activity of 3C protease involved in viral replication or maturation. Our findings demonstrate that A1AT treatment prevents a WCS-induced increase in viral load and for the first time suggest a therapeutic effect of A1AT on HRV infection.

Antiviral drug screening by assessing epithelial functions and innate immune responses in human 3D airway epithelium model

“…To study respiratory infections, traditionally cell lines and undifferentiated monolayer epithelial cells in submerged cultures were used, which couldn't replicate faithfully the real mechanisms of viral infections in human respiratory tract. Therefore, increasing number of studies turned recently toward ALI models, because of its high physiological and functional relevance, to investigate certain aspects of the respiratory viral infections (Berman et al, 2016;Farsani et al, 2015;Griggs et al, 2017;Jones et al, 2016). In this study, we undertook a rather holistic analysis of the physiological modifications happening in a human 3D airway ALI culture upon viral infection and revealed the importance of an innate immune response which is virus strain dependent.…”

“…Overall, our data support the use of the standardized MucilAir™ airway as a promising preclinical model for the design and the test of novel antiviral strategies. In comparison with other air-liquid interface 3D approaches (Berman et al, 2016;Farsani et al, 2015;Griggs et al, 2017;Jones et al, 2016), MucilAir™ is made of low passage human airway epithelial cells ensuring a maximum similarity with physiological condition and the reconstruction is standardized with quality control using functional tests. Some important assets of MucilAir™ for pre-clinical testing have already been addressed.…”

MUC18 Regulates Lung Rhinovirus Infection and Inflammation

“…Epithelial cells were washed off the trachea in a 2% Fetal Bovine Solution/DMEM medium and incubated for 4 hours to allow debris to adhere to the dish. Cells were then seeded onto transwell inserts and cultured for one week before being shifted to air liquid interface (ALI) as previously described [ 11 ]. On the 10 th day of ALI, cells were infected with HRV-1B at 5x10 4 50% Tissue Culture Infective Dose (TCID 50 ) as previously described and harvested 24 hours later.…”

The Utilization of Alpha-1 Anti-trypsin (A1AT) in Infectious Disease Monitoring and Treatment

“…Instead of binding and inactivating HRV 3C protease, which involves in HRV replication and maturation, A1AT suppresses HRV host receptor gene expression, namely, intercellular adhesion molecule-1 (ICAM-1) (). In turn, A1AT treatment on infected brushed bronchial epithelial cell culture reduces viral load [34].…”

“…They also imply administration of A1AT may help A1AT deficient infected patients to counteract the damage caused by the infection. [34] and anti inflammatory [35] Dengue virus high level indicates acute phase infection [19;20,21]; higher level in DF and DHF cases compared to healthy control. A trend of higher level as the severity degree increases but not significant [26] no reference available yet [42,44], prevents tissue damage [44], inhibits hemolysis [42] Mycobacterium abscessus predispose patient to develop NTBI [18] bacteria reduction [18] Mycobacterium tuberculosis increase in TB patients compared to control participant [22];decline at the end of TB therapy and burden disease [25] no reference available yet…”