Multi-parameter flow cytometric analysis of the functional profile of Vγ9Vδ2 T-cells.

<p>Representative dot plots of functional parameters (MIP-1β, CD107A, IFNγ panels B and D) and the respective Isotype control of antibodies (panels A and C) were shown in healthy donors (HD) and HIV-infected patients (HIV+) after stimulation with Picostim. For the gating strategy, after the lymphocyte population was gated sequentially, CD3+ and Vγ9Vδ2 T-cell events were defined, and gates for each respective function were made (panels B and D, light blue gates) using combinations that provided optimal separation. Single function gates were set based on the negative control (unstimulated) samples and were placed consistently across samples. We used the Boolean gate platform to create the full array of possible combinations, equating to 8 (2³) response patterns when testing three functions. Data are reported after background correction. Numbers on the light blue gates indicate the percentage of functional Vγ9Vδ2 T-cells among the total Vγ9Vδ2 T-cells (black boxes).