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

Journal: Frontiers in Immunology

Article Title: Discovery and Use of Long dsRNA Mediated RNA Interference to Stimulate Antiviral Protection in Interferon Competent Mammalian Cells

doi: 10.3389/fimmu.2022.859749

Figure Lengend Snippet: Primary Bronchial Epithelial/Tracheal Cells (pBECs) pre-soaked with long dsRNA of viral genes inhibits infection with corresponding viruses. The pBECs were grown to confluence and were shown to exhibit characteristics indicative of epithelial/tracheal cells, including mucous production and cilia function (A) . The pBECs were pre-soaked with either DPBS, 500 ng/mL of the mis-matched mCherry dsRNA control or 500 ng/mL of VSV N protein dsRNA before infection with VSV-GFP (MOI = 0.1) for 24h (B) . The pBECs were also pre-treated with either DPBS, 50 μg/mL of HMW pIC, 500 ng/mL of the mis-matched mCherry dsRNA control or 500 ng/mL of HCoV-229E M protein dsRNA before infection with HCoV-229E (MOI = 0.1) for 24h (C) . Error bars represent +SEM, and each data point represents the average of 3 independent replicates. A p-value of less than 0.05 was considered to be statistically significant. Error bars with different letters represent significantly different data.

Article Snippet: Normal human primary bronchial/tracheal epithelial cells (pBECs) were purchased from ATCC (PCS-300-010).

Techniques: Infection, Control