A Kinetic Study On Production Of Cytokines By Peripheral Blood Cells

Abstract

Cytokines are large group of low molecular weight that are polypeptides or glycopeptides in nature and are essential mediators of inflammation and immune reactions. The estimation of cytokine production after cell stimulation is used for understanding the immune status of an individual and thereby in diagnosis and prognosis of diseases. The kinetic study, on several cytokine at one platform, for production of cytokines is scanty. Hence, we have attempted a kinetic study (in terms of dose of stimulants for pro and anti inflammatory cytokines and their time of production) of various cytokines by whole blood cell culture. After stimulating blood cells with phytohaemagglutinin (PHA-L) or     lippolysaccharide (LPS) as stimulant, various cytokines {IL-1b, IL-12p70, IL-2, IL-4,      IL-10, and tumor necrosis factor alpha (TNF-α ) and interferon gamma (IFN-γ)} were estimated in cell culture supernatants by flow cytometry. We have observed that for production of IL-1b, IL-12p70,  IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ optimal concentration of PHA were 8µg/ml, 16µg/ml, 16µg/ml, 16µg/ml, 8µg/ml, 16µg/ml, 16µg/ml and 8µg/ml respectively. On the other hand, the optimal time for production of these cytokines were found to be 120 hours, 48 hours,  24 hours, 144 hours, 120 hours, 48 hours, 24 hours and 96 hours respectively. Regarding LPS as stimulant, the optimal concentration of LPS for production of IL-1b, IL-12p70, IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ were 8µg/ml, 8µg/ml, 8µg/ml, 8µg/ml, 16µg/ml, 4µg/ml, 4µg/ml and 4µg/ml respectively; whereas the optimal timings for production were 96 hours, 96 hrs, 48 hours, 48 hours, 96 hours, 48 hours, 24 hours and 120 hours respectively. These findings would be of relevance in understanding the status of cytokine production in health and diseases.