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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 investigate the effect of dendritic cells (DCs) derived from peripheral blood mononuclear cells (PBMCs) on the differentiation of Th1 and Th17 cells in normal pregnant women and preeclampsia patients. Methods The experimental group was 32 cases of preeclampsia and the control group was 20 cases of normal pregnant women. PBMC from normal pregnancy group and preeclampsia group were isolated and monocytes were adherent. GM-CSF, IL-4 and LPS were cultured to induce mature DCs. Flow cytometry was used to detect the expression of surface markers CD14, CD80, CD83 and CD86 The content of IL-23 in culture supernatant was detected by ELISA. The CD4 + T lymphocytes were sorted by magnetic beads and cultured with dendritic cells derived from peripheral blood mononuclear cells (N-DCs) of normal pregnant women. Cytokines IL-2 were also added together with peripheral blood mononuclear cells (PBMCs) of preeclampsia Monocyte-derived dendritic cells (P-DCs) and IL-2 are co-cultured or co-cultured with N-DC while adding cytokines IL-1β and IL-6 or co- Cytokines IL-1β, IL-6 were added. The above groups were cultured to detect the proportion of CD4 + IFN-γ + Th1, CD4 + IL-17 + Th17 cells by flow cytometry on the 6th day. Results The expression of CD83, CD80 and CD86 in P-DC was higher than that in N-DC, the difference was statistically significant (P <0.05). P-DC and different cytokines, CD4 + T cells to differentiate into Th1, Th17 cells higher than the ability of N-DC, the difference was statistically significant (P <0.01). Conclusion The changes of the phenotype and function of DC in peripheral blood of preeclampsia may be related to the immune imbalance of patients.