Decreased intra-lymphocyte cytokines measurement in septic shock patients: a whole blood proof of concept study

摘要: ABSTRACT Functional testing protocols are thought to be the gold standard for exploration of immune system. However, in terms routine analysis, they present numerous drawbacks and consequently their use is mainly limited research applications. In clinical context septic shock, characterized by marked lymphocyte alterations, a new approach intracellular cytokine measurement whole blood upon was evaluated proof-of-concept study. Following activation, simultaneous labeling Interferon-γ (IFN-γ), Tumor Necrosis Factor-α (TNF-α), Interleukin-2 (IL-2) performed CD4+ CD8+ T cells (identified surface marking). The analysis carried out flow cytometry (6 colors). Results obtained patients (n = 22) were compared those healthy volunteers 8). Independently lymphopenia, there significant differences between groups. particular decrease production IL-2 TNF-α patients, while IFN-γ not significantly altered. Polyfunctional results showed that presented with increased percentages triple negative lymphocytes. contrast, had higher proportions positive cells. could robust consistent way, taking 4.5 hours complete. Moreover, clear observed groups this modified method. These characteristics illustrate potential novel protocol further required determine applicability alternative test evaluate performances larger cohorts patients.