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PECULIARITIES OF CYTOKINE PRODUCTION BY LYMPHOCYTES UNDER VIRAL LOAD SIMULATION IN VITR-
Abstract
The authors have conducted an experiment that involved modeling of interactions between immune regulation mechanisms applying a viral load simulator in vitro. The experiment was performed on a suspension that contained immune cells extracted from peripheral blood samples (68 samples overall). There were several influencing factors such as impacts exerted by managing elements including cytokines (IL-1), hydrocortisone, and exogenous environmental factors represented by aluminum compounds (aluminum oxide) as well as a viral load simulator (VLS) which was a universal mitogen based on lipopolysaccharides. We analyzed effects in vitro as resulting from exposure to two various concentrations, the first one corresponding to a reference level, the second one being 2-10 times higher. Thus, hapten modification of the immune response was modeled via introducing aluminum oxide in concentrations being equal to 0,01 mg/l (the reference concentration for the substance) and 0,1 mg/l. Samples were exposed to various combinations of endo- and exogenous factors and the exposure lasted for 72 hours; after that we measured concentrations of cytokines that characterized post-induction cytokine production by lymphocytes population. We determined concentrations of IL-8, IL-10, IL-17 cytokines with Elx808IU analyzer applying ELISA technique and ?Vector-Best? test systems (Russia). All the obtained results were statistically analyzed with Statistica 6.0 applied software. Validity of discrepancies was assessed with Student?s t-test and discrepancies between groups were considered to be valid at ?<0,05. Our experiment results allowed us to reveal peculiarities of combined effects produced by endogenous and exogenous modifiers that influenced production of immune regulation mediators; these peculiarities indicated that the effects had additive nature. We examined peculiarities related to functioning of immunocytes exposed to a viral load simulator in in vitro experiment; the results revealed that immune response could vary and its variability was characterized with an increase in IL-8 and IL-10 production and a decrease in IL-17 production. Combined exposure to 10 pg/ml of IL-1 with endo- and exogenous modifiers (hydrocortisone and aluminum oxide) resulted in inhibited IL-8 and IL-10 expression and stimulated IL-17 expression against minimal viral simulation assessed as per a level of its biological analogues. In case higher IL-1 concentration (100 pg/ml) was introduced, it led to modification in immune response as an authentically enhanced effect produced by a simulator regardless of a modifier. We applied a mathematic model and it allowed us to establish that all detected regularities of influence exerted by immune-biologic and chemical factors with various origins on functions performed by immune-competent cells were non-linear ones. Detected peculiarities of induced pro- and anti-inflammatory cytokines expression can be used for efficient predicting and eliminating disorders in immune responses under various exposure, including combined one, to endogenous homeostatic factors and hapten environmental ones.
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