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Intensive Care Medicine Experimental
Fig. 1 | Intensive Care Medicine Experimental

Fig. 1

From: Intracellular calcium signaling and phospho-antigen measurements reveal functional proximal TCR activation in lymphocytes from septic shock patients

Fig. 1

Intracellular calcium signaling in CD4+ T cells from septic patients and controls upon TCR ligation. Peripheral blood mononuclear cells (PBMC) were isolated from patients and controls and loaded with Fluo-4AM for 30 min at 37 °C. Cells were analyzed by flow cytometry for 5 min at baseline then stimulated by biotinylated anti-CD3 (OKT3) antibody during 90 s before addition of streptavidin to induced TCR stimulation and analyzed for another 5 min. As a positive control, ionomycin was added at the end of experiment. Left, representative example of overtime intracellular calcium staining in one healthy volunteer. Right, intracellular calcium staining following TCR ligation in septic patients (n = 7) and healthy volunteers (controls, age-matched, n = 7) before (baseline), after TCR stimulation (biotinylated anti-CD3 antibodies + streptavidin) and after ionomycin addition. MFI of Fluo-4AM was measured in CD4+ T lymphocytes on 3 periods of 100 s at baseline and after complete TCR stimulation. For each stimulation condition, the maximum MFI was considered. Data are represented as means +/− SD. *p < 0.05 compared to baseline, Wilcoxon paired test

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