人CD200基因轉(zhuǎn)染未成熟樹突狀細胞體外誘導調(diào)節(jié)性T細胞的實驗研究
發(fā)布時間：2018-11-27 21:24
【摘要】： CD200是近年來新發(fā)現(xiàn)的免疫球蛋白超家族成員,研究證明CD200與其受體相互作用,可以下調(diào)髓樣細胞的活性、減輕自身免疫性疾病的癥狀、誘導免疫耐受等作用。1995年,Sakaguchi等發(fā)現(xiàn)了CD4~+CD25~+調(diào)節(jié)性T細胞(Tr),調(diào)節(jié)性T細胞可能是機體以“主動”方式來維持自身穩(wěn)定的主要途徑。Tr對免疫反應具有抑制效應,在體外增殖能力低,在免疫病理、移植物耐受、阻止自身免疫反應和維持機體免疫平衡方面都有一定作用。 本研究用脂質(zhì)體轉(zhuǎn)染的方法,將pcDNA3-CD200重組質(zhì)粒轉(zhuǎn)染到外周血分離的未成熟樹突狀細胞(imDC)內(nèi),用熒光標記的抗人CD200抗體作用后,經(jīng)流式細胞儀監(jiān)測CD200的表達情況。再用經(jīng)轉(zhuǎn)染CD200基因的未成熟DC誘導臍血淋巴細胞產(chǎn)生CD4~+CD25~+調(diào)節(jié)性T細胞(Tr),經(jīng)流式細胞儀檢測調(diào)節(jié)性T細胞的表型,為進一步探討CD200基因誘導機體免疫耐受機制的研究奠定基礎。 結果表明:pcDNA3-CD200重組質(zhì)粒轉(zhuǎn)染到未成熟DC后,CD200分子在DC表面大量表達,轉(zhuǎn)染CD200基因的未成熟DC誘導的產(chǎn)生CD4~+CD25~+調(diào)節(jié)性T細胞(Tr)明顯高于對照組;同時證明調(diào)節(jié)性T細胞(Tr)表面CD4~+CD25~+分子的表達與其表面FOXP3分子的表達在統(tǒng)計學上沒有顯著性差異。 本研究的意義在于:建立穩(wěn)定表達CD200分子的未成熟DC體系,轉(zhuǎn)染CD200基因的未成熟DC可以誘導產(chǎn)生更多的Tr,提示誘生Tr是CD200誘導免疫耐受的途徑之一;同時,在調(diào)節(jié)性T細胞中FOXP3的表達與CD4~+CD25~+分子的表達在統(tǒng)計學上差異沒有顯著性,且CD200分子可以誘生更多的Tr,從而推斷CD200基因具有誘導FOXP3分子表達的作用,為CD200誘導免疫耐受機制的進一步研究奠定了基礎。
[Abstract]:CD200 is a newly discovered member of immunoglobulin superfamily in recent years. It has been proved that the interaction between CD200 and its receptors can down-regulate the activity of myeloid cells, relieve the symptoms of autoimmune diseases, induce immune tolerance and so on. Sakaguchi et al found that CD4~ CD25~ regulatory T cell (Tr), regulatory T cell may be the main way for the body to maintain its own stability in an "active" manner. Tr has inhibitory effect on immune response, low proliferative ability in vitro and immunopathology. Graft tolerance, blocking autoimmune responses and maintaining immune balance play a role. In this study, pcDNA3-CD200 recombinant plasmid was transfected into (imDC) of immature dendritic cells isolated from peripheral blood by liposome transfection, and the expression of CD200 was monitored by flow cytometry after using fluorescent labeled anti-human CD200 antibody. Immature DC transfected with CD200 gene was used to induce CD4~ CD25~ regulatory T cell (Tr), in cord blood lymphocytes. Flow cytometry was used to detect the phenotype of regulatory T cells. It lays a foundation for further study on the mechanism of immune tolerance induced by CD200 gene. The results showed that after transfection of pcDNA3-CD200 recombinant plasmid into immature DC, CD200 molecules were expressed on the surface of DC. (Tr) of CD4~ CD25~ regulatory T cells induced by immature DC transfected with CD200 gene was significantly higher than that of control group. At the same time, there was no statistical difference between the expression of CD4~ CD25~ molecule on regulatory T cell (Tr) surface and that of FOXP3 molecule on regulatory T cell surface. The significance of this study lies in the establishment of immature DC system expressing CD200 molecules stably. Immature DC transfected with CD200 gene can induce more Tr, to induce Tr as one of the pathways of CD200 induced immune tolerance. At the same time, there was no statistically significant difference between the expression of FOXP3 and CD4~ CD25~ in regulatory T cells, and CD200 could induce more Tr, thus inferred that CD200 gene could induce the expression of FOXP3. It lays a foundation for the further study of the mechanism of immune tolerance induced by CD200.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R392
本文編號：2362074
[Abstract]:CD200 is a newly discovered member of immunoglobulin superfamily in recent years. It has been proved that the interaction between CD200 and its receptors can down-regulate the activity of myeloid cells, relieve the symptoms of autoimmune diseases, induce immune tolerance and so on. Sakaguchi et al found that CD4~ CD25~ regulatory T cell (Tr), regulatory T cell may be the main way for the body to maintain its own stability in an "active" manner. Tr has inhibitory effect on immune response, low proliferative ability in vitro and immunopathology. Graft tolerance, blocking autoimmune responses and maintaining immune balance play a role. In this study, pcDNA3-CD200 recombinant plasmid was transfected into (imDC) of immature dendritic cells isolated from peripheral blood by liposome transfection, and the expression of CD200 was monitored by flow cytometry after using fluorescent labeled anti-human CD200 antibody. Immature DC transfected with CD200 gene was used to induce CD4~ CD25~ regulatory T cell (Tr), in cord blood lymphocytes. Flow cytometry was used to detect the phenotype of regulatory T cells. It lays a foundation for further study on the mechanism of immune tolerance induced by CD200 gene. The results showed that after transfection of pcDNA3-CD200 recombinant plasmid into immature DC, CD200 molecules were expressed on the surface of DC. (Tr) of CD4~ CD25~ regulatory T cells induced by immature DC transfected with CD200 gene was significantly higher than that of control group. At the same time, there was no statistical difference between the expression of CD4~ CD25~ molecule on regulatory T cell (Tr) surface and that of FOXP3 molecule on regulatory T cell surface. The significance of this study lies in the establishment of immature DC system expressing CD200 molecules stably. Immature DC transfected with CD200 gene can induce more Tr, to induce Tr as one of the pathways of CD200 induced immune tolerance. At the same time, there was no statistically significant difference between the expression of FOXP3 and CD4~ CD25~ in regulatory T cells, and CD200 could induce more Tr, thus inferred that CD200 gene could induce the expression of FOXP3. It lays a foundation for the further study of the mechanism of immune tolerance induced by CD200.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R392
【參考文獻】
相關期刊論文 前2條
1 侯治富,郭楠,高申,鄭德明,卜麗莎,高嵩,才華;人外周血樹突狀細胞的誘導與鑒定[J];吉林大學學報(醫(yī)學版);2005年05期
2 李浩威,段連寧;樹突狀細胞誘導T細胞免疫耐受的機理[J];國外醫(yī)學(免疫學分冊);2003年05期
，本文編號：2362074
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