Department of immunology; School of Basic Medical Science; China Medical University; Shenyang, P.R. China.
Hum Vaccin Immunother. 2013 Apr;9(4):773-83. doi: 10.4161/hv.23137. Epub 2013 Mar 7.
To gain new insight into the functional interaction between dendritic cells and methionine encephalin (MENK) combined with pidotimod (PTD), we have analyzed the effect of MENK plus PTD on the morphology, phenotype and functions of murine bone-marrow derived dendritic cells (BMDCs) in vitro. The maturation of BMDCs cultured in the presence of either MENK or PTD alone, or MENK in combination with PTD, was detected. The cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt/phenazinemethosulphate (MTS/PMS). The changes of BMDCs morphology were confirmed with light microscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The BMDCs treated with MENK combined with PTD displayed a higher expression of typical maturation markers of CD40, CD80, CD83, CD86 and MHC-IIidentified by fluorescence activated cell sorting (FACS), and stronger ability to drive T cells. The decrease of the endocytic ability was assayed by DAB kit, FITC-dextran and cellular immunohistochemistry. Finally upregulation of cytokines production of IL-12 and TNF-α was determined by ELISA. These data indicate that MENK combined with PTD could exert synergistic action on BMDC maturation.
为了深入了解树突状细胞与蛋氨酸脑啡肽（MENK）结合匹多莫德（PTD）的功能相互作用，我们分析了 MENK 加 PTD 对体外培养的鼠骨髓来源树突状细胞（BMDCs）形态、表型和功能的影响。检测了单独用 MENK 或 PTD 或 MENK 与 PTD 联合培养的 BMDCs 的成熟情况。通过 3-（4,5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑，内盐/ phenazinemethosulphate（MTS/PMS）测量细胞增殖。用光镜、透射电镜（TEM）和扫描电镜（SEM）证实 BMDCs 形态的变化。用 MENK 联合 PTD 处理的 BMDCs 通过流式细胞术（FACS）显示出更高表达的典型成熟标志物 CD40、CD80、CD83、CD86 和 MHC-II，并且具有更强的驱动 T 细胞的能力。通过 DAB 试剂盒、FITC-葡聚糖和细胞免疫组织化学测定内吞能力的下降。最后通过 ELISA 测定细胞因子 IL-12 和 TNF-α的产生上调。这些数据表明，MENK 联合 PTD 可对 BMDC 成熟发挥协同作用。
