@article {Alkhoshaiban1066, author = {Abdulaziz Alkhoshaiban and Priyanka Anujan and Lynne Bingle and Colin Bingle}, title = {Using mouse tracheal epithelial cells (mTECs) to uncover an antiviral role for BPIFB1}, volume = {56}, number = {suppl 64}, elocation-id = {1066}, year = {2020}, doi = {10.1183/13993003.congress-2020.1066}, publisher = {European Respiratory Society}, abstract = {BPIFB1 is a secretory protein produced by the respiratory tract buts its{\textquoteright} function remains unresolved. By analogy to other members of the wider BPI-fold contain protein family it has been assumed to function in host defence. Function studies have been hampered by the fact that the gene is only expressed in differentiated primary airway cells grown at the air liquid interface (ALI). Influenza A virus (IAV) is a significant respiratory pathogen and our preliminary data showed that bpifb1 is reduced in lungs following IAV (X31) infection. We hypothesise that BFIFB1 contributes to the defence mechanism against IAV infections and plays a role in maintaining homeostasis within the airway epithelium. In this study we validated the use of primary mouse tracheal epithelial cells (mTECs) to study the role of BPIFB1 in IAV infection. Gene expression analysis of mTECs showed that bpifb1 steadily increases during 14 days of ALI differentiation. Western blotting confirmed production of BPIFB1 as an N-glycosylated protein. Inducing a {\textquotedblleft}mucous{\textquotedblright} phenotype in the cells by IL-13 treatment induced bpifb1. A series of FLAG-tagged expression clones including WT, an N-glycosylation mutant and proteins corresponding to the N- and C-terminal BPI domains were shown to produce recombination murine BPIFB1 proteins in transiently transfected HEK cells by Western blotting. We developed an IAV infection assay in mTECs and are currently undertaking studies to identify regions of BPIFB1 associated with viral infection. Our study has established mTEC cells as suitable model to study the role of BPIFB1 in viral infections and further work will use Bpifb1-/- mTECs cells to directly examine the role of BPIFB1 in IAV infection.FootnotesCite this article as: European Respiratory Journal 2020; 56: Suppl. 64, 1066.This abstract was presented at the 2020 ERS International Congress, in session {\textquotedblleft}Respiratory viruses in the "pre COVID-19" era{\textquotedblright}.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).}, issn = {0903-1936}, URL = {https://erj.ersjournals.com/content/56/suppl_64/1066}, eprint = {https://erj.ersjournals.com/content}, journal = {European Respiratory Journal} }