Abstract
Pretreatment of super-low-dose lipopolysaccharide (SL-LPS) induces a more hyperresponsive state on the production of proinflammatory mediators to a subsequent secondary challenge with high-dose LPS in innate immune cells. Low-dose glucocorticoids (GCs) are also known to induce inflammation and immunosuppression in the immune cells. However, there is limited knowledge on whether preconditioning of low-dose GCs enhances inflammatory responses and dysregulates T lymphocyte responses to secondary LPS in SL-LPS-primed immune cells. In the present study, RAW 264.7 and EL4 cells were pretreated with SL-LPS (50 pg/ml) or low-dose corticosterone (CORT50: 50 ng/ml and CORT100: 100 ng/ml) in fresh complete medium once a day for 2-3 days, consecutively, and then cultured in fresh complete medium for 6 or 24 h in the presence or absence of LPS (1-10 μg/ml) or concanavalin A (Con A). The results demonstrated that the repeated pretreatment of CORT50 strongly enhanced production of IL-6, IL-10, TNF-α, and nitric oxide (NO) by RAW 264.7 cells in EP (SL-LPS-primed cells: endotoxin priming) in the absence of LPS compared to those in control (vehicle-pretreated cells), whereas CORT100 reduced production of TNF-α and IL-10. Further, the repeated pretreatment of CORT50 markedly enhanced LPS-induced production of IL-6, IL-10, TNF-α, PGE(2), and NO by RAW 264.7 cells in EP compared to those in control, whereas CORT100 attenuated LPS-induced production of IL-6, IL-10, and NO. Moreover, the repeated pretreatments of CORT50 and CORT100 greatly attenuated the Con A-stimulated production of IFN-γ and IFN-γ/IL-10 and LPS-stimulated production of IL-10, IFN-γ, and IFN-γ/IL-10 by SL-LPS-primed EL4 cells (EP). These findings suggest that double preconditionings of low grade hypercortisolemia and metabolic endotoxemia may act as important risk factors for metabolic disorder and severe morbidity and mortality in septic shock via upregulated production of inflammatory mediators and immunosuppression of IFN-γ-mediated responses.