【摘要】： ApoE-/-小鼠CD4+CD25+ T細(xì)胞分離及功能檢測 目的探討ApoE-/-小鼠CD4+CD25+T細(xì)胞的比例和功能及其與動(dòng)脈粥樣硬化的關(guān)系。方法從apoE-/-小鼠外周血流式細(xì)胞儀(FCA)檢測CD4+CD25+T細(xì)胞的比例并分分選之;混合淋巴細(xì)胞反應(yīng)(MLR)研究CD4+CD25+T細(xì)胞的免疫抑制功能;ELISA法測定上清液中細(xì)胞因子IL-10、TGF-β濃度;觀察ApoE-/-小鼠動(dòng)脈粥樣斑塊的形成狀況。結(jié)果與正常小鼠相比,ApoE-/-小鼠外周血中CD4+CD25+T細(xì)胞數(shù)量無差異,但抑制naive T細(xì)胞增殖能力弱,分泌更少IL-10、TGF-β;ApoE-/-小鼠斑塊面積較大。結(jié)論ApoE-/-小鼠CD4+CD25+T細(xì)胞抑制功能弱,可能因此免疫失穩(wěn)而致動(dòng)脈粥樣硬化。 HSP60特異性調(diào)節(jié)性T細(xì)胞的誘導(dǎo)及其對小鼠動(dòng)脈粥樣硬化影響 目的探討抗原特異性CD4+CD25+T細(xì)胞的體外誘導(dǎo)及其對動(dòng)脈粥樣硬化斑塊形成的影響。方法從apoE-/-小鼠分離骨髓單個(gè)核細(xì)胞,經(jīng)雷帕霉素(RPM)處理培養(yǎng)出未成熟樹突狀細(xì)胞(DC);體外誘導(dǎo)HSP60特異性調(diào)節(jié)性T細(xì)胞分化;流式細(xì)胞儀(FCA)檢測及分選CD4+CD25+T細(xì)胞;混合淋巴細(xì)胞反應(yīng)(MLR)研究CD4+CD25+T細(xì)胞的特異性抑制效應(yīng);ELISA法測定上清液中細(xì)胞因子IL-10、TGF-β和IFN-γ濃度。過繼轉(zhuǎn)移CD4+CD25+T細(xì)胞后,觀察小鼠動(dòng)脈粥樣斑塊的形成狀況。結(jié)果雷帕霉素處理的DC共刺激分子CD80和CD86表達(dá)明顯減少,形態(tài)學(xué)表現(xiàn)為未成熟樹突狀細(xì)胞;未成熟樹突狀細(xì)胞比成熟樹突狀細(xì)胞能誘導(dǎo)更多CD4+CD25+T細(xì)胞;培養(yǎng)體系中細(xì)胞因子IL-10、TGF-β水平明顯升高;CD4+CD25+T細(xì)胞能抑制效應(yīng)性T細(xì)胞的增殖及IFN-γ的分泌。過繼HSP60特異性CD4+CD25+T細(xì)胞組小鼠斑塊面積較小。結(jié)論未成熟樹突狀細(xì)胞可誘導(dǎo)出抑制功能強(qiáng)大的HSP60特異性CD4+CD25+T細(xì)胞,后者在體內(nèi)能能明顯抑制斑塊的形成。
[Abstract]:Isolation and functional determination of CD4 CD25 T cells in ApoE-/- mice objective to investigate the proportion and function of CD4 CD25 T cells in ApoE-/- mice and their relationship with atherosclerosis. Methods the proportion of CD4 CD25 T cells was detected by flow cytometry (FCA) in peripheral blood of apoE-/- mice, and the immunosuppressive function of CD4 CD25 T cells was studied by mixed lymphocyte reaction (MLR). The concentration of cytokine IL-10,TGF- 尾 in supernatant was determined by ELISA method, and the formation of atherosclerotic plaques in ApoE-/- mice was observed. Results compared with normal mice, the number of CD4 CD25 T cells in peripheral blood of ApoE-/- mice was not different, but the ability of inhibiting the proliferation of naive T cells was weak, and the secretion of IL-10,TGF- 尾 was less, and the plaque area of ApoE-/- mice was larger. Conclusion the inhibitory function of CD4 CD25 T cells in ApoE-/- mice is weak, which may result in atherosclerosis due to immune instability. Induction of HSP60 specific regulatory T cells and their effects on atherosclerosis in mice objective to investigate the induction of antigen-specific CD4 CD25 T cells in vitro and their effects on atherosclerotic plaque formation. Methods Bone marrow mononuclear cells were isolated from apoE-/- mice and immature dendritic cells (DC);) were cultured with rapamycin (RPM) to induce HSP60 specific regulatory T cell differentiation in vitro. The specific inhibitory effect of CD4 CD25 T cells was studied by mixed lymphocyte reaction (MLR), and the concentration of cytokines IL-10,TGF- 尾 and IFN- 緯 in supernatant was determined by ELISA assay. After adoptive transfer of CD4 CD25 T cells, the formation of atherosclerotic plaques in mice was observed. Results the expression of CD80 and CD86 in DC costimulatory molecules was significantly decreased after treatment with rapamycin, and the morphological features were immature dendritic cells, and immature dendritic cells could induce more CD4 CD25 T cells than mature dendritic cells. The level of cytokine IL-10,TGF- 尾 was significantly increased in culture system, and CD4 CD25 T cells could inhibit the proliferation of effectual T cells and the secretion of IFN- 緯. The plaque area of adoptive HSP60 specific CD4 CD25 T cell group was smaller. Conclusion immature dendritic cells can induce potent HSP60 specific CD4 CD25 T cells which can inhibit plaque formation in vivo.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R392

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