本文選題：細(xì)胞因子 + 間充質(zhì)干細(xì)胞(MSC)��； 參考：《蘇州大學(xué)》2006年碩士論文

【摘要】： 目的:通過(guò)體外實(shí)驗(yàn)研究間充質(zhì)干細(xì)胞(mesenchymal stem cell,MSC)對(duì)于不同刺激條件下的T淋巴細(xì)胞亞群的免疫調(diào)節(jié)作用,從而探討間充質(zhì)干細(xì)胞移植對(duì)于移植后的移植物抗宿主病(GVHD)和移植物抗白血病(GVL)的發(fā)生所起的作用,旨在尋求間充質(zhì)干細(xì)胞與造血干細(xì)胞進(jìn)行聯(lián)合移植的優(yōu)勢(shì)和潛在應(yīng)用價(jià)值。 方法:通過(guò)Ficoll-Hypaque梯度密度離心法分離出正常人骨髓單個(gè)核細(xì)胞,體外培養(yǎng)擴(kuò)增MSC,獲取第3代細(xì)胞。將其按照不同的比例加入雙向混合淋巴細(xì)胞培養(yǎng)(mixed lymphocyte culture, MLC)體系中,分別在第1,3,5天,用MTT比色法檢測(cè)各組MLC中的T淋巴細(xì)胞的增殖情況,再用流式細(xì)胞術(shù)分析與MSC共孵育體系中或經(jīng)佛波脂(phorbol myristate acetate,PMA)及離子霉素(Ionomycin)共同刺激下的T淋巴細(xì)胞的表面標(biāo)記以及其內(nèi)因子IL-4和IFN-γ的分泌變化情況。 結(jié)果:加入了MSC的MLC體系中,MSC對(duì)T淋巴細(xì)胞的增殖抑制具有量效關(guān)系,且隨著共孵育時(shí)間延長(zhǎng),抑制程度增強(qiáng);其中,CD4+T細(xì)胞亞群受抑不如CD8+T細(xì)胞亞群顯著;另外,T淋巴細(xì)胞表面CD25的表達(dá)雖然較對(duì)照組有所下降,但是CD4CD25共表達(dá)的細(xì)胞卻較對(duì)照組有明顯的上升趨勢(shì)。T淋巴細(xì)胞表面活化抗原HLA-DR的表達(dá)較對(duì)照組有輕度減低。根據(jù)T細(xì)胞內(nèi)因子的變化,Th1和Tc1的比率較對(duì)照組有顯著降低,而Th2和Tc2的比率卻有輕度上升。于共刺激作用下單獨(dú)培養(yǎng)的T細(xì)胞,MSC能輕度抑制其Th1細(xì)胞的轉(zhuǎn)化;而在MLC中,MSC則能夠明顯抑制其中的CD8+T細(xì)胞亞群和Th1細(xì)胞,輕度提高Th2細(xì)胞比率。 結(jié)論: MSC在體外能夠明顯抑制T淋巴細(xì)胞的增殖,其主要是針對(duì)CD8+T細(xì)胞。另外,它還能下調(diào)活化T淋巴細(xì)胞上一些較特異的表面標(biāo)記CD25和HLA-DR的表達(dá),降低MLC體系中的Th1和Tc1,升高Th2和Tc2。所以在臨床上可能有減輕移植后急性移植物抗宿主病的發(fā)生,而保留移植物抗白血病的潛力。
[Abstract]:Objective: to investigate the immunomodulatory effect of mesenchymal stem cells (MSCs) on T lymphocyte subsets under different stimulation conditions in vitro. The purpose of this study was to explore the role of mesenchymal stem cell transplantation in the development of graft-versus-host disease (GVHD) and graft-versus-leukemia (GVLL) after transplantation, in order to explore the advantages and potential value of combined transplantation of mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs). Methods: normal human bone marrow mononuclear cells were isolated by Ficoll-Hypaque gradient density centrifugation. It was added into mixed lymphocyte culture, MLC) system in different proportion, and the proliferation of T lymphocytes in MLC was detected by MTT colorimetry on the 5th day of the first day of mixed lymphocyte culture. The surface labeling of T lymphocytes and the secretion of IL-4 and IFN- 緯 were analyzed by flow cytometry in MSC co-incubated system or stimulated by phorbol myristate acetatePMA and ionomycinin. Results: in the MLC system with MSC, the inhibition of T lymphocyte proliferation was dose-dependent, and the inhibition degree increased with the prolongation of co-incubation time, among which the suppression of CD8 T cell subsets was less significant than that of CD8 T cell subsets. In addition, the expression of CD25 on the surface of T lymphocytes was lower than that of the control group, but the expression of HLA-DR on the surface of T lymphocytes was slightly lower than that of the control group. The ratio of Th1 and Tc1 was significantly lower than that of the control group, but the ratio of Th2 and Tc2 was slightly increased. Co-stimulated T cells could slightly inhibit the transformation of Th1 cells, while in MLC, CD8 T cell subsets and Th1 cells were significantly inhibited, and the ratio of Th2 cells was slightly increased. Conclusion: MSC can significantly inhibit the proliferation of T lymphocytes in vitro, which is mainly directed at CD8 T cells. In addition, it can down-regulate the expression of CD25 and HLA-DR on activated T lymphocytes, decrease Th1 and Tc1 in MLC system, and increase Th2 and Tc2. Therefore, it may have the potential to reduce the occurrence of acute graft-versus-host disease after transplantation and preserve the potential of graft-versus-leukemia.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R392

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本文編號(hào)：1813938