雷帕霉素和地塞米松对小鼠树突状细胞分化成熟的调控

目的：观察雷帕霉素（rapamycin，Rap）和地塞米松（dexamethasone，Dex）对堵养的小鼠骨髓来源的树突状细胞（DC）分化发育的影响。方法：（1）用GM-CSF＋IL-4定向诱导C57BL／6小鼠骨髓细胞分化为DC，分别加入Rap或Dex，然后用脂多糖（LPS）刺激。在倒置显微镜和扫描电镜下，动态观察DC形态学E的变化。（2）通过流式细胞术（荧光抗体双标记法）测定CD11c^＋细胞的比例及CD86和MHC-Ⅱ类分子表达的变化。（3）通过单向混合淋巴细胞反应（MLR）观察，Rap和Dex处理的DC刺激BALB／c小鼠同种异基因T细胞增殖的情况。结果：（1）经Rap和Dex处理后，DC在形态学上呈现稳定不成熟状态。（2）Rap处理的细胞表面CDIlc和MHC-Ⅱ类分子的表达仅有轻度降低，而CD86的表达明显降低。Dex处理的细胞表面CDIlc的表达与Dex的剂量呈负相关，CD86和MHC-Ⅱ类分子的表达均明硅降低。两种药物处理的DC均可抵抗LPS的促成熟作用。（3）MLR的结果显示，Rap和Dex处理的DC刺激同种异基因BALB／c小鼠T细胞增殖的能力均较低。结论：Rap和Dex均可使DC处于稳定的不成熟状态。与Dex相比，Rap对骨髓造血F细胞向DC分化的过程影响较小，而且在抑制DC表面协同刺激分子CD86表达的同时，对MHC-Ⅱ类分子的表达影响较小。

Effect of rapamycin and dexamethasone on differentiation and maturation of murine bone marrow-derived dendritic cells

AIM: To observe the effect of rapamycin (Rap) and dexamethasone (Dex) on differentiation and development of murine bone marrow-derived dendritic cells (DC) in vitro. METHODS: DC cells generated from C57BL/6 murine bone marrow cells were induced by GM-CSF and IL-4. During the course, Rap or Dex was added to the culture and the cells were then stimulated by LPS. (1)Morphology development of DCs was observed by inverted microscope and scanning electron microscope.(2)The cells were analyzed by flow cytometry (FCM) to determine the proportion of CD11c(+) cells and the change of CD86 and MHC class II molecule.(3)The influence of DCs treated by Rap or Dex on the allogeneic T cell proliferation was studied by one-way MLR. RESULTS: (1)The morphology of DCs maintained in a durable state of immaturity after Rap or Dex pretreatment.(2)The expression of CD11c and MHC-II slightly decreased but CD86 dramatically reduced on Rap-treated DC cells. There was close relationship between the expression of CD11c on Dex-treated cells and the dosage of Dex. The surface expression of CD86 and MHC-II dramatically reduced on Dex-treated DCs. Moreover, DCs treated by Rap or Dex were both able to resist the maturation triggered by LPS.(3)Bone marrow-derived DCs cultured with Dex or Rap had a lower stimulatory effect on allogeneic T cells compared with that of mature DCs. CONCLUSION: Rap and Dex could keep DCs in durable immaturity. Compared with Dex, which inhibited the expression of co-stimulatory molecules, Rap hardly influenced the differentiation of DCs and the expression of MHC class II molecules.