Fig. 1 FACS analysis of the numbers of different subsets of
circulating CD3+CD4+T cells in KD
patients.
PBMCs were isolated from individual subjects and PBMCs
5*105/tube were stained in duplicate with
PE-Cy5-anti-CD3 and PerCP-anti-CD4 or isotype controls, fixed and
permeabilized, followed by intracellular staining with FITC-anti-IL-17,
PE-Cy7-anti-IFN-γ, and PE-anti-IL-22. The frequency of
CD3+CD4+IL-22+Th22,
CD3+CD4+IL-17+Th17
and
CD3+CD4+IFN-γ+Th1
cells were determined by flow cytometry analysis. The cells were gated
on living lymphocytes and then gated on
CD3+CD4+ cells, and at least about
30,000 events were analyzed for each sample. The numbers of each type of
CD3+CD4+IL-22+Th22,
CD3+CD4+IL-17+Th17
and
CD3+CD4+IFN-γ+Th1
cells were calculated, according to the total numbers of PBMCs, the
frequency of different types of
CD3+CD4+T cells. (A). Flow cytometry
analysis; (B). The numbers of
CD3+CD4+IL-22+Th22
cells; (C).
CD3+CD4+IL-17+Th17
cells; (D).
CD3+CD4+IFN-γ+Th1
cells. Data shown are representative FACS charts or the mean numbers of
each type of cells per ml of peripheral blood in individual subjects
from two separate experiments. The horizontal lines indicate the median
values for each group. Data shown are representative charts of different
subsets of CD3+CD4+ T cells from
individual groups of subjects (n=20 for the HC, n=17 for the patients
with CAL, and n=26 for the patients with NCAL).