RT Journal Article
SR Electronic
T1 Establishing ex vivo infection to influenza A virus in mouse precision cut lung slices
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP PA3691
DO 10.1183/13993003.congress-2021.PA3691
VO 58
IS suppl 65
A1 Julia Chitty
A1 Lovisa Dousha
A1 Philip Bardin
A1 Jane Bourke
A1 Belinda Thomas
YR 2021
UL http://erj.ersjournals.com/content/58/suppl_65/PA3691.abstract
AB 1. Precision cut lung slices (PCLS) offer a unique integrated experimental platform for investigating both inflammatory and immune responses to viral infection.2. This study aimed to develop a protocol for robust infection of PCLS ex vivo with influenza A virus (IAV) with a view to its application for assessment of antiviral agents.3. PCLS were prepared using agarose-inflated lungs from naïve C57Bl6 mice and infected with IAV (HKx31 mouse strain). Matched slices from each mouse at 3 IAV concentrations (1x104-1x106 PFU) were assessed 24 and 48 hr post infection (n=4-8). Viral loads were assessed by plaque assay of homogenised PCLS. Conditioned media was used to measure both cell death via LDH assay and inflammatory responses to infection via TNFα ELISA.4. A dose-dependent increase in viral load was observed at 24 hr, with a further 3-fold increase in plaque numbers at 48 hr at the lowest IAV concentration only (1x104PFU, p<0.05, paired t-test). LDH was increased at 48 hr, irrespective of viral load. TNFα levels in conditioned media were increased 5-fold between 24 and 48 hr with 1x104PFU IAV (57±11, 243± 56 pg/ml, p<0.01).5. Treatment of mouse PCLS with IAV ex vivo elicits dose- and time-dependent infection and release of inflammatory cytokines. Further studies assessing immune responses and sensitivity to viral treatment are required. PCLS may be a viable screening tool for pre-clinical assessment of mechanisms of infection and validation of new therapeutic targets.FootnotesCite this article as: European Respiratory Journal 2021; 58: Suppl. 65, PA3691.This abstract was presented at the 2021 ERS International Congress, in session “Prediction of exacerbations in patients with COPD”.This is an ERS International Congress abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).