Figure 3. 7-color, 9-markers immunophenotyping analysis scheme by flow cytometry.

The different leukocyte subtypes were analyzed by the shown gating strategy. In all CD45 positive leukocytes the following cells were determined: CD3 (lymphocytes); CD4 (Th lymphocytes); CD8 (Tc lymphocytes); CD19 (B lymphocytes). Leukocytes small differential picture was done using CD14 and SSC display. In lymphocytes CD3 positive and negative (non-T cells) have been identified. In T cells CD4 and CD8 positive cells were estimated. In non-T cells NK or B cells (based on CD16/56 or HLADR expression, respectively) were identified. Proportions of the analyzed cell types were determined and the respective absolute cell numbers of these cell types were calculated by the absolute cell counts.