摘要
目的探讨甘露聚糖结合凝集素（MBL）对脂多糖（LPS）刺激的不同分化和活化状态THP1/CD14细胞产生TNF-α和IL-12的影响.方法用PMA和/或IFN-γ诱导不同分化和活化状态的THP1/CD14细胞,以不同浓度人MBL预处理2 h后再用光滑型LPS刺激24h.收集培养上清,以ELISA从蛋白水平分析其TNF-α和IL-12 p40＋p70的产生.收集细胞提取总RNA,以RT-PCR从转录水平评估TNF-α和IL-12的表达.结果ELISA检测发现,LPS可刺激各组细胞分泌TNF-α和IL-12 p40＋p70;IFN-γ活化的THP1/CD14细胞产生IL-12 p40＋p70最高,PMA分化的THP1/CD14细胞最低;PMA分化＋IFN-γ活化的THP1/CD14细胞分泌TNF-α最高,未用PMA或IFN-γ预刺激的THP1/CD14细胞最低.高浓度MBL（50～100 mg/L）可抑制LPS诱导细胞分泌TNF-α和IL-12 p40＋ p70的作用,低浓度MBL（1～10 mg/L）则几无影响.RT-PCR分析亦显示,与相应只用LPS刺激的实验组相比,高浓度MBL（50 mg/L）对不同实验组LPS诱导的TNF-α、IL-12 p35和p40的mRNA表达均有不同程度的抑制作用.结论MBL可抑制LPS诱导的不同分化和活化状态THP1/CD14细胞产生TNF-α和IL-12,THP1/CD14细胞可用做进一步剖析MBL在免疫应答与细胞因子网络调控中的作用及其机理的模型.
Objective To investigate the effects of mannan-binding lectin （MBL） on TNF-α and IL-12 production induced by lipopolysaccharide （LPS） in differentiated and activated THP1/CD14 cells. Methods The differentiated and activated THP1/CD14 cells were induced by PMA and/or IFN-γ and were stimulated for 24 h with smooth LPS after pretreatment with human natural MBL at concentration ranging from 1 to 100 mg/L for 2 h. The content of TNF-α and IL- 12 p40 ＋ p70 in culture supernatants were detected by ELISA and the levels of TNF-α, IL-12 p35 and p40 mRNA expressions in these cells were determined by RT-PCR. Results ELISA showed that the secretion of TNF-α and IL-12 p40＋ p70 from THP1/CD14 cells in different states can be induced by LPS. The maximal amount of IL-12 p40＋ p70 was detected in the culture supematants of THP1/CD14 cells activated by IFN- γ and the nunimum in those of PMA-differentiated THP1/CD14 cells. The highest level of TNF-α was found in the culture supernatants of THP1/CD14 cells differentiated by PMA and activated by IFN-γ and the lowest in those of THP1/CD14 cells without PMA or IFN-γ stimulation . The productions of TNF-α and IL-12 p40 ＋ p70 by differently differentiated and activated THP1/CD14 cells induced with LPS are profoundly inhibited by MBL at higher concentrations （50-100 mg/L） but not MBL at lower concentrations （ 1-10 mg/L）. RT-PCR analysis also indicated that the mRNA expressions of TNF-α and IL-12 p35 and p40 subunits in THP1/CD14 cells decreased to various extents by MBL at higher concentration , compared to the corresponding THP1/CD14 cells stimulated with LPS alone. Conclusion MBL may inhibit TNF-α and IL-12 production induced by LPS from differentiated and activated THP1/CD14 cells and THP1/CD14 cells can be used as an appropriate cell model for further exploring the molecular mechanism of MBL in immune regulation and the signaling pathways involved in cytokines network.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第5期458-462,共5页
Chinese Journal of Microbiology and Immunology
