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.