The human 5637 BECs were seeded onto 24-well plates at a density of 2.5 × 105 cells per well and were infected with UPEC CI5 strain (AmpR) at MOI 200. After 1 h of infection, the extracellular bacteria were killed by gentamicin for an additional hour. The infected cells were then rinsed thoroughly, replaced with growth media, and incubated for 6 h at 37 °C with 5% CO2. When necessary, a human type I IFN neutralizing antibody mixture (PBL Assay Science) was used. Lactobacillus spp. were added at different MOI and incubated overnight at 37 °C. After removing media, BECs were treated with 0.1% Triton X-100 in phosphate-buffered saline (PBS) to lyse the cells and plated on MacConkey agar plates (BD) with carbenicillin.
Human type 1 ifn neutralizing antibody mixture
Manufactured by PBL Assay Science
The Human type I IFN neutralizing antibody mixture is a laboratory reagent designed to neutralize type I interferons (IFNs) in human samples. It is a mixture of antibodies that specifically bind to and inactivate human type I IFNs, including IFN-α, IFN-β, and IFN-ω. This product can be used in research applications to study the role of type I IFNs in various biological processes.
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2 protocols using human type 1 ifn neutralizing antibody mixture
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Invasion and Infection of Bladder Epithelial Cells
2
Interactions between Plasmacytoid Dendritic Cells and B Cells in ZIKV Infection
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For co-culture experiments, pDCs and B cells were individually isolated from healthy volunteers, using negative immunomagnetic selection procedures (Miltenyi Biotech). Subsequently, pDCs and B cells were cultured in 96-well plates at pDC: B-cell ratios of 0.4:1, and infected with ZIKV at a MOI of 1. At defined time points after infection, B cells infected in the presence or absence of pDCs were subjected to immunophenotyping by flow cytometry, using the following antibodies: CD19 (FITC, clone HIB19), CD123 (BV510, clone 6H6), CD303 (PerCP, clone 201A), CD69 (BV605, clone FN50), in addition to staining with BVD for isolation of live cells. When indicated, a human type I IFN neutralizing antibody mixture (PBL Assay Science) which effectively neutralizes biological activity of human IFN-α, IFN-β, IFN-Ω, IFN-κ, and IFN-ε was added. Class C CpG oligonucleotides (ODN 2395, InvivoGen), a TLR9 ligand, served as a positive control to activate B cells. In selected cases, isolated B cells (1 × 105 /well) and pDC (0.4 × 105) were co-cultured using the upper and lower compartments of a transwell culture system (0.4 μm, Costar, Wilmington, MA), respectively; co-culture of isolated T cells and pDC under similar conditions was used as a control. All flow cytometry data was analyzed using FlowJo v10 (FlowJo, USA).
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