新HH方对HBV转基因小鼠脾脏源DC相关的免疫耐受状态影响的研究

R esearch of New HH Recipe on the State of Immune Tolerance Related to Spleen DC in HBV Transgenic Mouse

目的:探讨新HH方对乙型肝炎病毒(Hepatitis B Virus,HBV)转基因小鼠脾脏源树突状细胞(Dendritic cell,DC)相关的"免疫耐受"状态影响。方法:取新HH方治疗过和未治疗过的转基因小鼠脾脏来源的单个核细胞,以重组小鼠粒细胞-巨噬细胞集落刺激因子(recombinant Mouse Granulocyte Macrophage-Colony Stimulatory Factor,rmGMCSF)和重组小鼠白细胞介素-4(Recombinant Mouse IL-4,rmIL-4)培养诱导DC,流式细胞仪检测其表面共刺激分子(CD80、CD86),MTT法检测其刺激同种异体淋巴细胞增殖的能力,酶联免疫法(ELISA)法测定培养上清液中白介素-12(Interleukin-12,IL-12)、白介素-10(Interleukin-10,IL-10)水平。结果:新HH方组小鼠DC的表面协同刺激分子(CD 86)表达水平(13.7±3.1)、刺激同种异体淋巴细胞增殖能力(1:50和1:10 SI分别为1.22±0.09、1.69±0.10)和细胞因子IL-12水平(256.72±29.75)均高于对照组(154.69±11.32)(P<0.05),细胞因子IL-10水平(45.32±11.28)均低于对照组(76.79±16.26)(P<0.05)。结论:新HH方可以促进HBV转基因小鼠DC的成熟,上调其表面协同刺激分子CD86的表达,增强了其促淋巴细胞细胞增殖和分泌IL-12的能力,加强其抗原递呈能力,抑制分泌IL-10,从而打破DC相关的免疫耐受状态。

To investigate the impact of New HH Recipe on the state of immune tolerance related to spleen DC in HBV transgenic mouse.Methods: Getting mononuclear cells from transgenic mice spleen,including treated and untreated by new HH Recipe,Cultured and induced DC by recombinant Mouse Granulocyte Macrophage-Colony Stimulatory Factor(rmGM-CSF) and Recombinant Mouse IL-4(rm IL-4),detected its costimulatory molecules(CD80,CD86) by flow cytometry,detected the ability to stimulate the proliferation of allogeneic lymphocytes by MTT assay,detected the level of culturing supernatant interleukin-12(IL-12) and interleukin-10(IL-10) by enzyme-linked immunosorbent assay(ELISA).Results: In New HH Recipe group,mouse DC surface costimulatory molecules(CD86) expression levels(13.70±3.10),and stimulate allogeneic lymphocyte proliferation(1: 50 and 1: 10 respectively 1.22±0.09,1.69±0.10) and cytokines IL-12 level(256.72±29.75) were higher(154.69±11.32)(P < 0.05),cytokine IL-10 levels were lower than the controlled group(45.32±11.28)(76.79± 16.26)(P <0.05).Conclusion: The New the HH Recipe can promote DC maturation of HBV transfected gene mice,raise its surface costimulatory molecule CD86 expression,enhance promoting lymphocyte proliferation and the ability to secrete IL-12,strengthen his antigen-presenting ability,inhibit the secretion of IL-10,so breake the state of immune tolerance DC-related.