Journal: PLoS ONE

Article Title: Dobrava-Belgrade Hantavirus from Germany Shows Receptor Usage and Innate Immunity Induction Consistent with the Pathogenicity of the Virus in Humans

doi: 10.1371/journal.pone.0035587

Figure Lengend Snippet: Vero E6 cells were treated with 40 µg/ml of indicated blocking antibodies for 1 hour. Then virus at multiplicity of infection 0.05 was added to the cells. After one hour cells were washed and new medium was added. One day later samples were collected. A ) Viral S-segment RNA was measured by qPCR. B ) Expression of viral nucleocapsid (N) protein was detected by Western blot. The density of bands on blots was quantified by ImageJ 1.41o programm (Wayne Rasband National Institutes of Health, USA). The percentages of antibody-mediated inhibition of viral infection were calculated in comparison to untreated but infected cells. Experiment was performed three times. Data are presented as the mean ± SD of the mean. C ) Representative picture of the Western blot analyses summarized in part B. —, untreated cells; β1, cells pre-treated with β1 integrin specific monoclonal antibody (MAb); β3, cells pre-treated with β3 integrin specific MAb; DAF, cells pre-treated with DAF specific MAb. D ) Binding of GRW/Aa to CHO cells stably expressing β3 integrins (CHO-β3cells) in comparison to control CHO cells. Virus binding was performed at 4°C for 1 hour. HTNV and PHV were used as controls. The amount of bound virus was measured through detection of viral RNA by specific qPCR. The binding affinity of virus particles to CHO-β3 cells is expressed as a ratio between virus genome equivalents detected on CHO-β3 cells and on the control CHO cells. Error bars represent standard deviations of the means from three experiments.

Article Snippet: Vero E6 (African green monkey epithelial kidney cell line; C1008 ATCC CRL 1586) and A549 cells (human epithelial lung cell line; ACC 107, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) were cultured in Minimal essential medium (MEM) with Earle's Salt supplemented with 5% fetal calf serum, 25 mM HEPES, 1% glutamine, 1% sodiumpyruvate, 1% non essential amino acids, and 0.1% gentamycine sulphate.

Techniques: Blocking Assay, Virus, Infection, Expressing, Western Blot, Inhibition, Comparison, Binding Assay, Stable Transfection, Control

Journal: PLoS ONE

Article Title: Dobrava-Belgrade Hantavirus from Germany Shows Receptor Usage and Innate Immunity Induction Consistent with the Pathogenicity of the Virus in Humans

doi: 10.1371/journal.pone.0035587

Figure Lengend Snippet: A ) 100 µl of indicated Vero E6-derived virus stocks or medium (NC, negative control) were exposed to UV irradiation. Amount of IFN-λ1 present in virus stocks was measured by ELISA. Abbreviation “ucf” means that the virus stock was purified by the ultracentrifugation procedure. Data are presented as the mean between three independently prepared stocks ± SD from the mean B ) GRW/Aa virus stock (∼50 µl) or recombinant proteins (rhIFN-λ1 or rhIFN-β) were preincubated with 1 µg of corresponding blocking antibody (anti-IFN-λ1 or anti-IFN-β). One hour after incubation the mixture was added on the top of A549 cells. Four days post infection MxA mRNA expression was measured by qPCR. NC (negative control), untreated A549 cells. Experiment was performed three times. Data are presented as the mean ± SD of the mean.

Article Snippet: Vero E6 (African green monkey epithelial kidney cell line; C1008 ATCC CRL 1586) and A549 cells (human epithelial lung cell line; ACC 107, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) were cultured in Minimal essential medium (MEM) with Earle's Salt supplemented with 5% fetal calf serum, 25 mM HEPES, 1% glutamine, 1% sodiumpyruvate, 1% non essential amino acids, and 0.1% gentamycine sulphate.

Techniques: Derivative Assay, Virus, Negative Control, Irradiation, Enzyme-linked Immunosorbent Assay, Purification, Recombinant, Blocking Assay, Incubation, Infection, Expressing

Journal: bioRxiv

Article Title: TMPRSS2 and TMPRSS4 mediate SARS-CoV-2 infection of human small intestinal enterocytes

doi: 10.1101/2020.04.21.054015

Figure Lengend Snippet: (A) MA104 cells in serum-free media were infected with SARS-CoV and SARS-CoV-2 chimera viruses (MOI=0.1) with or without trypsin (0.5 μg/ml) for 48 hours. (B) Human ileum enteroids seeded into collagen-coated 96-well plates were apically infected with SARS-CoV-2 chimera virus (MOI=0.1) for 24 hours. (C) Human ileum enteroids in 3D Matrigel were infected with SARS-CoV-2 chimera virus (MOI=0.1) for 24 hours and stained for virus (green), ACE2 (red), and nucleus (DAPI, blue). Also see Supplementary Video 1 for 3D view.

Article Snippet: African Green Monkey kidney epithelial cell lines MA104 (CRL-2378.1) and human embryonic kidney cell line HEK293 (CRL-1573) were originally obtained from American Type Culture Collection (ATCC) and cultured in complete M199 medium and complete DMEM medium, respectively.

Techniques: Infection, Virus, Staining

Journal:

Article Title: Extended Analysis of the In Vitro Tropism of Porcine Endogenous Retrovirus

doi:

Figure Lengend Snippet: Relative susceptibility of nonhuman cell lines to infection by PERV-NIH-2°-derived PERV pseudotypes

Article Snippet: 293 human embryonic kidney (ATCC CRL-1573), MMK Mus musculus molossinus kidney (ATCC CRL-6439; no longer available), and SC-1 mouse embryo epithelial (ATCC CRL-6450) cells, MDTF Mus dunni tail fibroblasts (kindly provided by Olivier Danos), NRK normal rat kidney cells (ATCC CRL-6509), Rat-2 rat embryo fibroblasts (ATCC CRL-1764), SIRC rabbit corneal fibroblasts (ATCC CCL-60), D17 canine oseosarcoma (ATCC CRL-6248), MDBK bovine kidney epithelium (ATCC CCL-22), FRhK4 rhesus monkey kidney (ATCC CRL-1688), Mv1Lu mink lung epithelial (ATCC CCL-64), and MiCL.1 (S+L−) mink lung (ATCC CCL-64.1) cells, Fc3TG feline tongue fibroblasts (ATCC CCL 176), and AK-D feline lung epithelial (ATCC CCL-150), CRFK feline kidney epithelial (ATCC CCL-94), CaKi-1 clear cell kidney carcinoma (ATCC HTB-46), HeLa cervical adenocarcinoma (ATCC CCL-2), HOS human osteosarcoma (ATCC CRL-1543), and CaCO-2 colorectal adenocarcinoma (ATCC HTB-37) cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine, 1 mM sodium pyruvate, 100 U of penicillin per ml, and 100 μg of streptomycin per ml.

Techniques: Infection, Control