Anti iga pe

CH12F3 cells were transduced with VLP HA-Vpr or ΔVpr. 24 h later, transduced cells were stimulated to undergo IgM-to-IgA switching with 5 ng/mL mouse recombinant IL4 (Peprotech), 200 ng/mL anti-CD40 monoclonal antibody (clone HM40-3, eBiosciences) and 1 ng/mL hTGF-β (R&D Systems) for 3 days. After labeling cells with anti-B220-APC (clone RA3-6B2, eBiosciences) and anti-IgA-PE (Southern Biotechnology), CSR efficiencies (% of IgGA⁺ B220⁺ cells) were evaluated by flow cytometry. Primary mature mouse B-cells were transduced with Vpr-delivering VLPs or control VLPs, and stimulated 24 h later to undergo IgM-to-IgG1 switching with 40 μg/mL LPS (Salmonella typhimurium, R&D Systems) and 50 ng/mL IL-4 (PeproTech) for 3 days. After labeling with anti B220-APC (clone RA3-6B2, eBiosciences) and anti-IgG1-PE (clone A85-1, BD Biosciences), CSR efficiencies (% of IgG1⁺ B220⁺ cells) were evaluated by flow cytometry. Data were acquired on a BD LSRII Fortessa cytometer and analyzed with the BD FACSDiva 6.1.3 software. Cell proliferation was measured with standard MTT/formazan colorimetric assay at 550 nm [35 (link)].