Fig. 2

Gating strategy for immune cell analysis. Whole blood was diluted with PBS and stained with antibodies. Granulocytes were sorted from the granulocyte area and selected for neutrophils by using antibodies against CD45, CD11b and Ly6G. The percentage of neutrophils was calculated from the whole leukocytes. Lymphocytes were sorted from the lymphocyte area by expression of CD45. B cells were identified using against CD19 antibodies. The percentage of B cells from CD45-positive cells in the lymphocyte gate was calculated. Various T cell populations were selected using antibodies against CD3 and specific markers. The percentage of the cytotoxic T cells (as identified by CD8 expression) and the total T helper cell population (as identified by CD4 expression) from total CD3-expressing T cells was calculated. The percentages of Th1, Th17 and Tregs from CD4-expressing helper T cells were determined using the surface markers as specified in the figure