Abstract
Activation of lymphocytes can effectively produce a large amount of cytokines. The types of cytokines produced may depend on stimulating reagents and treatments. To find an optimal method to stimulate cytokine production and evaluate its effect on immunotoxicity assessments, the authors analyzed production of IL-2, IL-4, IL-6, IL-10, IL-13, IFN-γ, TNF-α, GM-CSF, RANTES and TGF-β in undiluted rat whole blood culture (incubation for 0, 2, 4, 6, 8 or 10 h) with different concentrations of PMA/ionomycin, PHA, Con A, LPS and PWM. We also evaluated the effects of cyclosporin A and azathioprine on cytokine production. The results revealed a rapid increase of IL-2, IFN-γ, TNF-α, RANTES and TGF-β secretion within 6 h after stimulation with 25 ng/mL PMA and 1 μg/mL ionomycin. The inhibition of these cytokine profiles reflected the effects of immunosuppressants on the immune system. Therefore, the results of this is study recommend the detection of cytokine profiles in undiluted whole blood stimulated 6 h with 25 ng/mL PMA and 1 μg/mL ionomycin as a powerful immunotoxicity assessment method.
Highlights
IntroductionCytokines are small molecular weight proteins or peptides secreted by many cell types ( immune system cells) that regulate the duration and intensity of the immune response
Cytokines are small molecular weight proteins or peptides secreted by many cell types that regulate the duration and intensity of the immune response
The results showed that each stimulant induced production of a unique cytokine profile (Figure 1)
Summary
Cytokines are small molecular weight proteins or peptides secreted by many cell types ( immune system cells) that regulate the duration and intensity of the immune response. Cytokines participate in many physiological processes, including the regulation of immune and inflammatory responses. Cytokines released by CD4+ T cells at the onset of an immune response are thought to be decisive for pathological or physiological consequences [1]. Effects on cytokines can be analyzed at the protein level, including direct detection of the relative concentration of various cytokines in the circulation following experimental treatment, and in activated peripheral blood mononuclear cells (PBMCs) or diluted whole blood. These assays have (apparently) been used successfully in many instances, there are a number of major problems. Cytokines generally act primarily at a local level and that they are rapidly cleared from the circulation
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