PRO- AND ANTI-INFLAMMATORY CYTOKINES (TNF-Α, IL-6, IL-4, IL-10) SECRETION FROM PBMC

To observe the pro- and anti-inflammatory cytokine (TNF-α, IL-4, IL-6, IL-10) secretion from cultivated peripheral blood mononuclear cells (PBMCs) for 48h. Blood samples were obtained 42 women patients of Nadezhda Women’s Health Hospital after signing written informed consent. For PBMC isolation, 9 ml heparinised whole each patient processed by density gradient centrifugation using Pancoll (P04-60100, Pan-Biotech) 25 minutes at 400G. The buffy coat PBMCs collected washed with phosphate buffer saline. A total 5x106 /ml cultured in 0.5 RPMI medium (R-8758, Sigma) supplemented 1% HSA (GHSA-125, LifeGlobal) 48 hours 37ºC. Concentrations tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), interleukin-4 (IL-4) interleukin-10 (IL-10) media measured sandwich enzyme-linked immunosorbent assay (ELISA) (CSB-E04638h, CSB-E04740h, CSB-E04633h CSB E04593h, Cusabio Technology, respectively) according to manufacturers’ instructions. Results are presented as mean ±SD range. Descriptive statistics one-way ANOVA post hoc LSD performed SPSS Software ver. 21. P<0.05 was considered significant. No cytokines detected clear control. presence IL-10 TNF-α confirmed all samples. proinflammatory (IL-6 TNF-α) concentrations culture 48h cultivation 90.48±89.1 pg/ml (range: 25.05 – 266.43) 510.36±66.81 400.59 565.10), respectively. (IL-4 secreted 438.82±130.27 227.53 - 599.19) 4667.33±3505.7 1808.31 9964.99), Additionally significantly higher comparison other analysed (p<0.001). this study showed that secrete detectable amounts media. Furthermore supernatant had amount studied (IL-4, IL-6 TNF- α).