内吗啡肽1抑制人外周血来源树突状细胞成熟和功能

Endomorphine-1 inhibits maturation and functions of human peripheral bloodderived dendritic cells

目的研究加入脂多糖(LPS)刺激后内吗啡肽1(EM-1)对外周血来源树突状细胞(PBDC)的表型、细胞因子分泌和刺激T细胞增殖等方面的影响,探讨EM-1调节DC功能的可能作用途径及机制。方法用重组人粒细胞巨噬细胞集落刺激因子(rh GM-CSF)和重组人白细胞介素4(rh IL-4)联合刺激经培养和离心处理后的PBDC,分组加入LPS或EM-1刺激,观察细胞生长形态变化;流式细胞术检测PBDC表面CD80、CD86、CD83、HLA-DR、CC趋化因子受体7(CCR7)的水平;ELISA检测PBDC分泌的IL-10、IL-12、γ干扰素(IFN-γ)的水平;羟基荧光素二乙酸盐琥珀酰亚胺脂(CFSE)染色检测EM-1诱导后PBDC对T细胞增殖的影响。结果 EM-1使LPS刺激诱导后PBDC表面CD80、CD86、CD83、HLA-DR、CCR7表达下调;EM-1抑制LPS诱导成熟的PBDC释放炎性因子IL-12和IL-10,抑制PBDC对T淋巴细胞的增殖反应能力,减少PBDC和T淋巴细胞共培养上清液中IL-12和IFN-γ的分泌。结论 EM-1可抑制PBDC的成熟和功能。

Objective To investigate the effects of endomorphine-1（ EM-1） on the functional features of lipopolysaccharide（ LPS）-stimulated peripheral blood-derived dendritic cells（ PBDCs）,including PBDC phenotypes,cytokine profile and its capability of promoting T cell proliferation. Methods PBDCs were isolated from peripheral blood and differentiated in the presence of recombinant human granulocyte-macrophage colony-stimulating factor（ rh GM-CSF） and interleukin-4（ rh IL-4）.Then the cells were further activated by LPS or EM-1. Several surface markers were detected by flow cytometry including CD80,CD86, CD83, HLA-DR and CCR7. ELISA was used to detect cytokine levels of IL-10, IL-12, IFN-γ in the supernatants of the cultured PBDCs. Using co-culture of T lymphocytes and PBDCs stimulated by EM-1,the proliferation of T cells induced by PBDCs was determined by 5,6-carboxyfluorescein succinimidyl ester（ CFSE） staining. Results EM-1down-regulated the expressions of CD80,CD86,CD83,HLA-DR and CCR7 on the activated PBDCs. Moreover,EM-1decreased the secretion of IL-12 and IL-10 in these PBDCs. In co-culture of T lymphocytes and EM-1-treated PBDCs,T cell proliferation was impaired,and less IL-12 and IFN-γ were produced in the culture supernatant. Conclusion EM-1 could inhibit the maturation and immune functions of PBDCs.