Interleukin-1β (IL-1β), IL-1 receptor antagonist, and TNFα production in whole blood

Abstract

The ability of an individual to mount defense responses to infection depend in part on the capacity to produce cytokines such as interleukin 1 (IL-1) and tumor necrosis factor (TNF). The specialized equipment, labor intensity, and sterile practice required for the standard in vitro evaluation of cytokine production can make such evaluation impractical in some clinical situations. We report a method for stimulating whole blood to produce cytokines that can be implemented in laboratories without tissue culture facilities and requires minimal sample preparation. IL-1 beta and TNF alpha production in whole blood samples was stimulated with endotoxin and/or phytohemagglutinin in standard EDTA-containing vacuum collection tubes. After incubation, plasma was removed and frozen for later assay. Comparison of this whole blood method with isolated mononuclear cell cultures indicated a significant correlation for IL-1 beta production (r = 0.746, P = 0.005). This technique also produced the newly described cytokine, IL-1 receptor antagonist. We conclude that the whole blood method is an acceptable alternative to isolated cell culture methods for measuring IL-1 beta in situations that preclude the standard in vitro approach.

Pro‐ and anti‐inflammatory cytokines in healthy volunteers fed various doses of fish oil for 1 year

“…The relative simplicity of the whole-blood stimulation may render this method less prone to experimental biases: manipulation of the cell population studied is minimal, stimulation is started immediately upon removal of the cells from the circulation and experimental conditions can easily be kept constant. Moreover, Nerad et al [21] have shown a good correlation between whole blood IL-1b measurement and the method in which mononuclear cells are separated by density gradient centrifugation and stimulated ex vivo. Therefore, whole blood stimulation may represent a reflection of cytokine production capacity that is at least as relevant as a method using cell separation techniques.…”

“…IL-1b, TNF-a and IL-1Ra were determined by radioimmunoassay as described previously [19][20][21]. The intra-assay variability was less than 10%.…”

Detection of Human P2X7 Nucleotide Receptor Polymorphisms by a Novel Monocyte Pore Assay Predictive of Alterations in Lipopolysaccharide-Induced Cytokine Production

“…Whole blood samples were used to measure the cytokine responses in vitro after 6 or 24 h of stimulation with 0 or 100 ng/ml S. typhimurium LPS, where the former time point produces halfmaximal responses in most donors for many cytokines (43)(44)(45)(46). Additionally, LPS-stimulated platelets in whole blood are the source for abundant levels of endogenous adenine nucleotides (47) and the interaction between LPS-stimulated platelets and monocytes has been shown to augment the production of both TNF-␣ and IL-1␤ (48).…”

Proinflammatory Effects of IL-10 During Human Endotoxemia

“…We used whole blood stimulation rather than stimulation of isolated cells because the former system is considered to mimic in vivo conditions best, containing hormones, cytokines, and other soluble factors that may influence cytokine production (30). This method has been adopted by many different groups, and has been used extensively to study the regulation of cytokine production (19,37,38,(45)(46)(47)(48)(49). In the whole blood experiments, IL-10 induced a strong dose-dependent inhibition of LPS-induced IFN-␥ production, which was associated with a concurrent reduction in IL-12 and IL-18 concentrations.…”