title: Optimal method to stimulate cytokine production and its use in immunotoxicity assessment.
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.
authors.affiliation: Chinese Academy of Inspection and Quarantine, Beijing 100123, China. chenhm@aqsiqch.ac.cn
authors.forename: Wenchao
authors.initials: W
authors.lastname: Ai
authors.forename: Haishan
authors.initials: H
authors.lastname: Li
authors.forename: Naining
authors.initials: N
authors.lastname: Song
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Huiming
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D001379:Azathioprine / Q000494:pharmacology; D001773:Blood Cells / Q000187:drug effects* / Q000276:immunology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D016207:Cytokines / Q000276:immunology*; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D015759:Ionomycin; D008070:Lipopolysaccharides / Q000494:pharmacology; D037121:Plant Lectins / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D016207:Cytokines; D007166:Immunosuppressive Agents; D008070:Lipopolysaccharides; D037121:Plant Lectins; D015759:Ionomycin; D016572:Cyclosporine; D001379:Azathioprine