2.3 Flow cytometry analysis of Th1, Th2, Th17 and HSCs
T-helper 1 (Th1), T-helper 2 (Th2), T-helper 17 (Th17) and HSCs in bone
marrow of children and controls were detected by flow cytometry. The
anti-human antibodies (BD Pharmingen) included
APC-H7-CD3, APC-CD4, FITC-CD183,
PE-Cy7-CD196, FITC-CD34, PE-Cy7-CD38,PE-Cy7-CD45RA,PE-CD90, Alexa
Fluor 647-CD49f,PerCP-Cy5.5-CD3,PerCP-Cy5.5-CD56, PerCP-Cy5.5-CD11 and
PerCP-Cy5.5-CD19. Th1, Th2 and Th17
are the lineages of
CD3+CD4+CD183+CD196-,
CD3+CD4+CD183-CD196-and
CD3+CD4+CD183-CD196+respectively. Long-term HSCs (LT-HSCs) are the lineages presenting
lin-CD34+CD38-CD45RA-CD90+CD49f+,
while short-term HSCs (ST-HSCs) presenting
lin-CD34+CD38-CD45RA-CD90-CD49f-.
The monocyte was collected from bone marrow aspirates or peripheral
blood with a Ficoll density gradient. Cells were analyzed by a
FACSCalibur (BD Biosciences). Data analysis was performed using FlowJo
software.