Project description:Proteomics and phosphoproteomics measurments of human a lung epithelial cell line (Calu-3 cells) infected with the novel coronavirus SARS-CoV-2 over a time-course of 4,8 and 12 hours.
Project description:We are interested in having the gene microarray done on untreated and PMA treated (10 nM x 24 hr) human and mouse epithelial cells lines including CALU-3, MLE-15, primary bronchial human airway epithelial cells. We are especially interested in seeing whether sialyl or sulfo transferases, and MUC genes, are present and if they are induced by PMA. We hope to use this information to optimize future experiments designed to identify Siglec lung ligands. The lab is interested in studying untreated and PMA treated (10 nM x 24 hr) human and mouse epithelial cells lines including CALU-3, MLE-15, primary bronchial human airway epithelial cells. With special interest in understanding whether sialyl or sulfo transferases, and MUC genes, are present and if they are induced by PMA.
Project description:SARS-CoV-2, the virus responsible for COVID-19, infects both human airway epithelial cells and trigeminal ganglia. We assessed the consequences of SARS-CoV-2 infection on gene expression in Calu-3 cells and in primary Mus musculus trigeminal ganglia cells (mmTG). Here, we provide datasets that include raw reads and mapped reads for the following: 1) mock infected mmTG 48 hrs post infection; 2) SARS-CoV-2 infected mmTG 48 hrs post infection; 3) mock infected Calu-3 cells 24 hrs post infection; 4) SARS-CoV-2 infected Calu-3 cells 24 hrs post infection; 5) mock infected Calu-3 cells 48 hrs post infection; 6) SARS-CoV-2 infected Calu-3 cells 48 hrs post infection.
Project description:The mucus secreted by airway epithelial cells plays a key role in the protection against and clearance of particles and pathogens. In this project, a 3D model of the bronchial epithelium was established using Calu-3 cells grown on porous inserts at the air-liquid interface. The secreted proteins were characterized in long-term cultures and compared to the apical secretome of primary normal human bronchial epithelial (NHBE) cells. The apical secretome was collected and characterized in the Calu-3 model at day 4, day 11, and day 18 after air-liquid interface and in the NHBE model at day 4, day 12, and day 18 after air-liquid interface. "This project received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no 760928 (BIORIMA)."
Project description:Human bronchial epithelial cell line Beas-2B were infected with Streptococcus pneumoniae at Multiplicity of Infection (MOI) of 0.5 and 1 or treated with lipoteichonic acid (LTA) for 9 and 16 h. The mRNA profile changes upon infection shall be determined to investigate Streptococci pathogenesis.
Project description:We are interested in having the gene microarray done on untreated and PMA treated (10 nM x 24 hr) human and mouse epithelial cells lines including CALU-3, MLE-15, primary bronchial human airway epithelial cells. We are especially interested in seeing whether sialyl or sulfo transferases, and MUC genes, are present and if they are induced by PMA. We hope to use this information to optimize future experiments designed to identify Siglec lung ligands.
Project description:This series contains 5 groups of U133A arrays with targets from Calu-3 cell line post-infection with P aeruginosa strains. Calu-3 human lung epithelial cells were seeded onto 6-well plates and grown to about 80% confluency. Epithelial monolayers were infected with 108 CFUs per ml of each bacterial strain for 60 min and then washed extensively with phosphate-buffered saline. The cultures were subsequently incubated in fresh RPMI 1640 medium supplemented with 10% fetal bovine serum and 100 µg/ml gentamicin. After 6 h, total RNA from the cells was prepared with RNAzol and further purified with RNEasy for microarray hybridization. All arrays processed and globally scaled to 500 using MAS 5. CONTROL (4 arrays) - no infection; FRD1 (4 arrays); FRD1234 (3 arrays); FRD440 (4 arrays); FRD875 (4 arrays)
Project description:We used two model systems to characterise the changes caused by SARS-CoV-2 using quantitative proteomics. We used primary human epithelial cells differentiated at the air-liquid interface, first describing the relative abundance of proteins found in each major cell type of the epithelium, and then characterised the infection of ciliated cells specifically as these showed the greatest susceptibility to SARS-CoV-2 infection. We also used a lung epithelial cell line, CaLu-3, and compared the infection with B.29 and B.1.1.7 (Alpha) variant over a time-course of infection. In each case, immunostaining for nucleoprotein combined with FACS allowed for the purification of infected cells from uninfected ‘bystander’ cells.