子痫前期患者外周血树突状细胞对Th1/Th17细胞分化的影响

Effect of dendritic cells on the differentiation of Th1/Th17 in peripheral blood from preeclampsia patients

目的观察正常妊娠妇女和子痫前期患者外周血单个核细胞(PBMC)来源的树突状细胞(DC)对Th1、Th17细胞分化的影响。方法实验组为子痫前期疾病患者32例;对照组为正常妊娠妇女20例。分离正常妊娠组和子痫前期组PBMC,经贴壁获得单核细胞,加GM-CSF,IL-4和LPS培养诱导为成熟DC,流式细胞术检测表面分子标志CD14,CD80,CD83,CD86的表达,ELISA检测培养上清液中IL-23的含量。磁珠分选出CD4+T淋巴细胞,与正常妊娠孕妇外周血单核细胞来源的树突状细胞(N-DC)共同培养,同时添加细胞因子IL-2,或与子痫前期患者外周血单核细胞来源的树突状细胞(P-DC)、IL-2共同培养;或与N-DC共同培养,同时添加细胞因子IL-1β、IL-6;或与P-DC共同培养,并添加细胞因子IL-1β、IL-6。以上各组培养到第6天用流式细胞术检测CD4+IFN-γ+Th1、CD4+IL-17+Th17细胞比例。结果 P-DC中CD83、CD80、CD86的表达高于N-DC,差异有统计学意义(P<0.05)。P-DC与不同的细胞因子共同作用,促使CD4+T细胞分化为Th1、Th17细胞的能力高于N-DC,差异有统计学意义(P<0.01)。结论子痫前期患者外周血DC表型和功能的改变可能与患者免疫失衡有一定关系。

Objective To observe the effect of dendritic cells （DCs） derived from peripheral blood monouclear cells （PBMCs） on the differentiation of Th] and Th]？ in normal pregnancy women and preeclampsia patients. Methods PBMCs were obtained from 32 preeclampsia patients and 20 normal pregnancy controls, respectively. Then DCs were sorted from peripheral blood monocytes cultured in the presence of cytokines （GM-CSF, IL-4） and LPS for 8 d. The phenotypes of DCs （CD14, CD80, CD83, CD86） were detected by flow cytometry （FCM）. The content of IL-23 in the supernatant was detected by ELISA. CD4 ＋T lymphocytes were separated using the magnetic BD IMag Cell Separation System according to the manu- facturer＇s instructions. Purified CD4 ~T lymphocytes were clutured with mature DCs derived from normal pregnancy women （N-DC） and IL-2, or with mature DCs derived from preeclampsia patients （P-DC） and IL-2, or with N-DC and IL-l[5, IL-6, or with P-DC and IL-I[3, IL-6. At 6 days after culture, CD4 ＋ IFN-y~ T（Thl） and CD4 ~ IL-17 ~ T（Th]？） subsets were deter- mined by FCM. Results Compared with N-DC, P-DC expressed the higher levels of CD83, CD80, CD86 and manifestated the stronger ability of promoting the differentiation of CD4 ＋ T into Thl/Thl7 when cultured with different cytokines （ P 〈 O. 01 ）. Conclusion The changes in phenotype and function of DCs might be related to immune imbalance and be an important reason for preeclampsia.