Stanilova Spaska A, Dobreva Zlatka G, Slavov Emil S, Miteva Lyuba D
Department of Molecular Biology, Immunology and Genetics, Faculty of Medicine, Trakia University, Armeiska 11 St., 6000 Stara Zagora, Bulgaria.
Int Immunopharmacol. 2005 Apr;5(4):723-34. doi: 10.1016/j.intimp.2004.12.003.
The immunomodulatory properties of bioactive agents include the ability to induce cytokine production by the activated target cell. The effect of immunomodulatory C3 binding glycoprotein isolated from Cuscuta europea on the induction of human PBMC cytokine synthesis and the cell viability was investigated. Isolated PBMC from healthy donors were cultured for 24 h with C3bgp. We also studied the influence of C3bgp on the cytokine production in LPS, PHA, PWM and Dex treated PBMC. The quantitative determination of TNF-alpha, IL-12, IL-6 and IL-10 was performed in culture supernatant by ELISA. Results obtained demonstrated that C3bgp induced proinflammatory and immunoregulatory cytokine production, in the highest degree IL-12, followed by IL-6 and in lower degree TNF-alpha. IL-12 quantity was significantly increased in C3bgp stimulated cultures in comparison with LPS, PHA and PWM stimulated PBMC. C3bgp also increased IL-12 in PHA or PWM stimulated cultures, but not in LPS stimulated culture. C3bgp significantly increased IL-6 production compared to the PHA and PWM but not to LPS stimulation. On the other side, C3bgp inhibited IL-10 production after LPS, PHA and PWM stimulation. Cell viability in C3bgp stimulated cultures retained on the same level from 72 to 120 h of culturing, in contrast to LPS and PHA stimulated cultures. Based on the results presented, we conclude that the C3bgp exhibited immunomodulatory properties on the human PBMC. The ability of PDTC and Dex to down-regulate the effect of C3bgp on the proinflammatory cytokine production suggests that a part of the mechanism of action of C3bgp is mediated through NF-kB signal transduction pathway.
生物活性剂的免疫调节特性包括激活的靶细胞诱导细胞因子产生的能力。研究了从欧洲菟丝子中分离出的免疫调节性C3结合糖蛋白对人外周血单个核细胞(PBMC)细胞因子合成诱导及细胞活力的影响。从健康供体分离的PBMC与C3bgp一起培养24小时。我们还研究了C3bgp对LPS、PHA、PWM和地塞米松处理的PBMC中细胞因子产生的影响。通过ELISA对培养上清液中的TNF-α、IL-12、IL-6和IL-10进行定量测定。获得的结果表明,C3bgp诱导促炎和免疫调节细胞因子的产生,其中IL-12诱导程度最高,其次是IL-6,TNF-α诱导程度较低。与LPS、PHA和PWM刺激的PBMC相比,C3bgp刺激的培养物中IL-12的量显著增加。C3bgp还增加了PHA或PWM刺激培养物中的IL-12,但在LPS刺激的培养物中没有增加。与PHA和PWM相比,C3bgp显著增加了IL-6的产生,但与LPS刺激相比没有增加。另一方面,C3bgp抑制LPS、PHA和PWM刺激后IL-10的产生。与LPS和PHA刺激的培养物相比,C3bgp刺激的培养物中的细胞活力在培养72至120小时内保持在同一水平。根据所呈现的结果,我们得出结论,C3bgp对人PBMC表现出免疫调节特性。PDTC和地塞米松下调C3bgp对促炎细胞因子产生影响的能力表明,C3bgp的部分作用机制是通过NF-κB信号转导途径介导的。