Fig. 2.

Overview of used fluorochrome-conjugated antibodies (Beckman Coulter)

Gating strategy: Leukocyte count is gated by CD45-KrO antibody and side scatter parameters (SSC). Lymphocyte protocol: gated as CD45⁺ cells with low side scatter. B-cells are CD3⁻CD19⁺ in the next scattergram and T-cells as CD3⁺CD19⁻. Tcr-gd subsets are CD3⁺ and TCRgd-positive in the next scattergram. In a following scattergram T-regulatory lymphocytes are identified using CD25-PC5.5 antibody and CD127-PE antibody as CD25⁺CD127dim.

Monocyte protocol: gated as CD14⁺ with low SSC. Monocytic subpopulations are subdivided using CD16-PC7 antibody and CD14-PE antibody to classical monocytes CD14++CD16⁻, intermediate monocytes CD14++CD16⁺ and non-classical monocytes CD14⁺CD16++.