酵母多糖影响人朗格汉斯细胞C型凝集素受体的研究

The Impact of Zymosan on C-Type Lectin Receptors on Human Langerhans Cells

探究酵母多糖(Zymosan)对人朗格汉斯细胞(Langerhans cells,LCs)C型凝集素受体(C-type lectin receptors,CLRs)的影响。收集健康志愿者血液样本,Ficoll淋巴细胞分离液分离外周血单个核细胞,CD14磁珠分选CD14+细胞,用含有GM-CSF(1 000 U/m L)、IL-4(1 000 U/m L)、TGFβ-1(10 ng/m L)的培养液诱导培养LCs,Zymosan刺激LCs,8、24 h后收集细胞。提取Zymosan实验组和对照组细胞RNA,实时RT-PCR分析5种CLRs(Dectin-1、Dectin-2、DC-SIGN、Mincle、MRC-2)mRNA的水平,扩增产物测序鉴定。共检测10个个体来源LCs在酵母多糖刺激后其CLRs mRNA变化的情况。在10组样本中,有6组Dectin-2受体mRNA较对照组增加(变化倍数≥2),其中3组增加明显(变化倍数≥5),最大增加倍数为20.91;有5组Mincle受体mRNA较对照组增加(变化倍数≥2),其中4组增加明显(变化倍数≥5),最大增加倍数为19.98;有1组MRC-2受体mRNA较对照组明显增加(变化倍数≥5)。Dectin-1和DC-SIGN受体mRNA没有增加。Zymosan能增加部分个体来源LCs内Dectin-2、Mincle和MRC-2受体mRNA的水平,而对Dectin-1和DC-SIGN受体mRNA的水平没有影响。本研究结果完善了Zymosan调节机体免疫功能的作用机制。

The impact of zymosan on C-type lectin receptors （CLRs） on human Langerhans cells （LCs） was investigated. Blood samples from healthy volunteers were collected, and PBMCs were isolated using Ficoll lymphocyte isolating liquid, and CD14 cells were isolated using CD14 microheads. The LCs were induce-cultured in medium liquid containing GM-CSF （1 000 U/mL） , IL-4 （1 000 U/mL） , TGFβ-1 （10 ng/mL） ; LCs were stimulated with zymosan and collected the cells after 8 h and 24 h. RNA was extracted from zymosan group and control group, then the levels of 5 CLRs （ Dectin-1, Deetin-2, DC-SIGN, Mincle, MRC-2 ） mRNA were analyzed by real-time RT-PCR, the ampli- fied products were identified by sequencing. The variances of CLRs mRNA of a total 10 individuals LCs after stimula- ted with zymosan were determined and tested. The results showed that among the 10 samples, 6 groups Dectin-2 receptor mRNA increased as compared with the control group （variance times 〉12）, among them 3 groups significantly increased （variance times ≥5） , the largest increasing times was 20.91; 5 groups Mincle receptor mRNA increased as compared with the control group （ variance times ≥ 2）, among them 4 groups significantly increased （ variance times ≥5）, the largest increasing times was 19.98; 1 group MRC-2 receptor mRNA significantly increased as conl- pared with the control group （variance times ≥ 5 ）. Dectiu-1 and DC-SIGN receptor mRNA were not increased. Therefore, zymosan can increase part of individual mRNA levels of Dectin-2, Mincle and MRC-2 receptor mRNA in LCs and ilas no effect on mRNA levels of the Dectin-1 and DC-SIGN receptor. This study perfected the mechanism of zymosan on regulating immune response.