Flow-Cytometry gating strategy for analysis of blood B-cell populations of HIV-1 infected individuals.

<p>Representative plot showing gating strategy on 105 live PBMCs. Total CD19+ B-cells were selected based on expression of CD27 and/or IgM, and levels of CD21. CD1c and CD10 expression were used for further characterization of blood MZ and TI B-cell populations. Quadrants were set based on the expression values obtained with fluorescence minus one (FMO) and isotype controls. Mature activated B-cells were defined as CD19+CD27+IgM-CD21loCD1c-CD10-, resting switched memory B-cells were CD19+CD27+IgM-CD21hiCD10-, precursor marginal-zone (MZ)-like B-cells were CD19+CD27+IgM+CD21loCD1c+CD10+, mature MZ-like B-cells were CD19+CD27+IgM+CD21hiCD1c+CD10- and transitional immature (TI) B-cells were CD19+CD27-IgM+CD21hiCD1c-CD10+. The mean events gated were: total B-cells (9320 ± 1750), mature activated (360 ± 67), resting switched memory (632 ± 301), precursor MZ-like (145 ± 36) mature MZ-like (327 ± 233) and TI (944 ± 174).