2.3 Measurement of serum sCD25 and cytokine levels
Peripheral blood samples were collected from LCH patients at the time of
initial diagnosis and one week after treatment in available patients.
Serum samples were separated by centrifugation at 1000 g for 20 min
within four hours of blood collection. Processed samples were either
immediately analyzed or stored at −80˚C until use. Serum levels of sCD25
were measured in duplicated using a specific Enzyme-linked immunosorbent
assay (ELISA; Kindstar Globalgene Technology Inc.). Serum cytokine
levels of interferon-γ (IFN-γ), tumor necrosis factor α (TNF-α), IL-10,
IL-6, IL-4, IL-2 were determined by Cytometric Bead Array (Human Th1/Th2
Cytokine Kit II, BD Biosciences), as described
previously31.