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. 2021 Oct 28;2:184–193. doi: 10.1016/j.crimmu.2021.10.002

Fig. 3.

Fig. 3

Means (±standard deviation) of the percentage of TNF-α producing T cells and monocytes detected in whole blood samples (n = 5). Fig. 3A shows the proportion of TNF-α producing monocytes after stimulation with PMA + Ionomycin (PO), SEB + CD28 (SEB) or LPS. Fig. 3B shows the percentage of TNF-α, IFN-γ and IL-6 producing T cells and monocytes in whole blood samples following stimulation with PMA and Ionomycin (for CD3+Cytokine + subsets) and with LPS (for CD14+cytokine + subsets). Fig. 3C is for IL-4, IL-2 and TGF-b whereas Fig. 3D shows the IL-12p40 and IL-12p70 producing T cells and Monocytes. 50 μl of stimulated and unstimulated whole blood samples were labelled in eight FACS tubes at a ratio of 1:25 for anti-CD3-PerCP for the analysis of T cells and at a ratio of 1:50 for anti-CD14-APC. After permeabilisation, the following cytokines, all PE-conjugated: TNF-α (BD FastImmune, Clone 6401.111), IFN-γ (BD FastImmune, Clone 25723.11), IL-10 (BD Pharmingen, Clone JES3-9D7), IL-6 (BD Pharmingen, Clone MQ2-13A5), IL-4 (BD FastImmune, Clone 3010.211), IL-2 (BD Pharmingen, Clone MQ1-17H12), TGF-β (BD Pharmingen, Clone TW4-2F8), IL-12 p40 (BD Pharmingen, Clone C11.5) and IL-12 p70 (BD Pharmingen, Clone 20C2) were added anti-cytokine-PE at a ratio of 1:12.5 before flow cytometric analysis.