2000
DOI: 10.1016/s0022-1759(00)00230-1
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Flow cytometric measurement of intracellular cytokines

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Cited by 194 publications
(131 citation statements)
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“…The same splenocytes used to assess T cell proliferation were used for detection of Th1 (IFN␥) or Th2 (IL4) lymphokines by ELISPOT (BD PharMingen) as described previously Agadjanyan et al, 2005). In addition, the FACS method was used for detection of specific cytokines by CD4 ϩ T cells (Pala et al, 2000). Briefly, the cultures of splenocytes from experimental and control animals were restimulated for 3-4 d with PADRE peptide (5 M), and then for 4 -6 h with PMA (phorbol 12-myristate 12-acetate) and ionophore (ionomycin) (both from Sigma, St. Louis, MO).…”
Section: Methodsmentioning
confidence: 99%
“…The same splenocytes used to assess T cell proliferation were used for detection of Th1 (IFN␥) or Th2 (IL4) lymphokines by ELISPOT (BD PharMingen) as described previously Agadjanyan et al, 2005). In addition, the FACS method was used for detection of specific cytokines by CD4 ϩ T cells (Pala et al, 2000). Briefly, the cultures of splenocytes from experimental and control animals were restimulated for 3-4 d with PADRE peptide (5 M), and then for 4 -6 h with PMA (phorbol 12-myristate 12-acetate) and ionophore (ionomycin) (both from Sigma, St. Louis, MO).…”
Section: Methodsmentioning
confidence: 99%
“…Where intracellular cytokine staining was measured, 10 mg of brefeldin A/ml (secretion inhibitor) was added to the cell cultures after 1-h stimulation with the different treatments to block the secretion of cytokines (21). Cells were incubated at 37˚C under 5% CO 2 overnight.…”
Section: Seb Stimulation Of Human Cells In Vitromentioning
confidence: 99%
“…After isolation, one aliquot of PBMC was stained for the detection of cell-surface antigens, while another aliquot was transferred to culture flasks and stimulated with 10 ng/ml phorbol 12-myristate 13-acetate (Sigma) and 1 mM ionomycin (Sigma), in the presence of 1 mM monensin (Sigma) for 4 h at 37 8C in 5% CO 2 (12,13).…”
Section: Intracellular Cytokine Detectionmentioning
confidence: 99%