primary bronchial epithelial cells  (ATCC)

Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: IGF2BP3 is increased in type 2 disease. A. Left: normalized expression levels of IGF2BP3 mRNA in severe asthma (SA) vs healthy control (HC) primary BECs at total and polyribosome-bound levels; centre: IGF2BP3 normalized microarray expression data comparing airway epithelial cells from healthy non-atopic (HNA) and atopic asthma (AA) children; right IGF2BP3 normalized microarray expression data comparing nasal lavage samples from children during Picornavirus-induced exacerbation (EX) vs 7-14 days post-exacerbation (post-EX). B. Normalized count levels comparing type 2 high vs type 2 low nasal brushings for genes that showed IGF2BP3-dependent IL-13-modulation in our Frac-seq data . C: Heatmap showing the normalized expression values of the common mRNA signature between differentially expressed genes in the GALA II cohort and our Frac-seq IGF2BP3 and IL-13-dependent DEGs. D. Model for IGF2BP3 effects on type 2 responses. Decreasing IGF2BP3 levels leads to increased stability of IL13RA1 and IL4R mRNA and increased surface expression of IL4R and IL13Rα1, leading to increased STAT6 phosphorylation and increased IL-13 (and possibly IL-4) transcriptional responses, contributing to an overall increased type 2 response. In disease, IGF2BP3 is upregulated potentially to supress excessive T2 responses.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Expressing, Control, Microarray, Phospho-proteomics

Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: Decreasing IGF2BP3 levels increases IL-13/IL-4-driven transcriptional responses. RT-qPCR results of primary BECs transfected with siRNAs against IGF2BP3 (siIGF2BP3) or scrambled control (siControl) and stimulated with IL-13 or IL-4. A: RT-qPCR results for IGF2BP3 knock down and the major signalling mediators of IL-13 and IL-4. B: RT-qPCR results for chemokines. C: RT-qPCR results for pro-inflammatory IL6 and IL8 mRNAs. Data (n=3) depicted as mean ±SEM. One-tailed ratio t-tests. * P ≤ 0.05; ** P ≤ 0.01.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Quantitative RT-PCR, Transfection, Control, Knockdown, One-tailed Test

Journal: Virology Journal

Article Title: In-vitro renal epithelial cell infection reveals a viral kidney tropism as a potential mechanism for acute renal failure during Middle East Respiratory Syndrome (MERS) Coronavirus infection

doi: 10.1186/1743-422X-10-359

Figure Lengend Snippet: SARS- and MERS-CoV receptor expression and virus infection experiments in human primary cells. (A) SARS- and MERS-CoV receptor expression of human Angiotensin-converting-enzyme-2 (ACE-2) and Dipeptidyl-peptidase-4 (DPP-4), respectively, in cryosections of a healthy human kidney and (B) in primary bronchial (HBEpC) and renal (HREpC) epithelial cells. For positive controls, ACE2- and DPP-4-expressing primate cells (kidney cells from African green monkey [Vero E6]) were stained in parallel. Cell lines known to be negative for ACE-2 (kidney cells from Syrian hamster [BHK]) or DPP-4 (kidney cells from African green monkey [COS-7]) were used as negative controls. The white bar represents 50 μm. (C) Cell morphology and cytopathic effect (CPE) formation of human primary renal epithelial cells (HREpC) infected with MERS-CoV or SARS-CoV with 0.5 plaque-forming units of either virus per cell. A pronounced CPE formation 20 hours post infection (hpi) was seen only after infection with MERS-CoV in HREpC but not in cells infected with SARS-CoV. No CPE formation was seen in HBEpC infected with MERS-CoV or SARS-CoV (data not shown). Upper row: 100-fold magnification, lower row: 400-fold magnification, bright field microscopy (D) Replication of SARS- and MERS-CoV on HBEpC and HREpC determined by real time RT-PCR after 0, 20 and 40 hpi (E) Progeny virus measured by titration of supernatants in duplicates in a plaque assay in Vero cells. MERS-CoV replicates in HREpC with peak titers of 6.2 log plaque forming units (PFU)/mL, showing a 2,9-fold log difference between HREpC and HBEpC, while replication of SARS-CoV showed only a 1-fold log difference between bronchial and renal primary cells. Replication levels for each virus used are given as log of the genome equivalents (GEs) (D) or as plaque-forming units (PFUs) (E) . All virus infection experiments were performed in triplicates. Bars represent mean values, error bars represent standard deviation of triplicates.

Article Snippet: As the bronchoalveolar epithelium of the lung constitutes the primary target compartment for both viruses, infection of human primary bronchial epithelial cells (HBEpC, Promocell, Heidelberg) was studied in parallel.

Techniques: Expressing, Infection, Staining, Microscopy, Quantitative RT-PCR, Titration, Plaque Assay, Standard Deviation