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. 2019 Jul 25;216(10):2331–2347. doi: 10.1084/jem.20190164

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© 2019 Bancroft et al.

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Figure 3. — Identification of human B cells able to bind anti-iglb12 idiotypes. (A) Detection of live CD19⁺ CD3⁻ CD14⁻ CD16⁻ B cells from human PBMCs that bound a cocktail containing IB1-APC, IB2-APC, and IB3-APC tetramers in fractions enriched or depleted of APC⁺ cells using anti-APC microbeads before flow cytometry. APC755 tetramers containing isotype control antibodies were included in these experiments to exclude B cells specific for the APC, streptavidin, and conserved portions of the anti-idiotypes. The displayed plots were derived from ∼100,000 APC-depleted or ∼400,000 APC-enriched cells derived from 100 million PBMCs and representative of three similar experiments. The mean percentage ± SD of IB1/2/3-APC⁺ APC755⁻ B cells in the enriched and depleted fractions from three individuals is shown on the plots. (B) The frequency of ten antibodies cloned from IB1/IB2/IB3-APC⁺ APC755⁻ B cells from an individual were assayed for binding to IB1, IB2, IB3, or IB5 by BLI. Each antibody was assessed for binding in two to three independent experiments.