本文選題：可溶性HLA-G二聚體 + 樹(shù)突狀細(xì)胞　； 參考：《華中科技大學(xué)》2013年碩士論文

【摘要】：HLA-G是一種非經(jīng)典的HLAⅠ類分子，可以通過(guò)與抑制性受體結(jié)合誘導(dǎo)廣泛的免疫耐受。大量研究證實(shí)可溶性HLA-G二聚體與其抑制性受體的結(jié)合力更強(qiáng)，可以誘導(dǎo)更強(qiáng)的免疫耐受。這些抑制性受體包括ILT-4和ILT-2，主要表達(dá)在T細(xì)胞，B細(xì)胞，巨噬細(xì)胞，NK，DC等表面。DC是重要的抗原提呈細(xì)胞，連接了固有免疫與適應(yīng)性免疫，它們可以發(fā)現(xiàn)、捕獲，加工處理外源性抗原并提呈給T細(xì)胞。它們與原始T細(xì)胞的相互作用在誘導(dǎo)T細(xì)胞分化成Th1/Th2細(xì)胞、調(diào)節(jié)性T細(xì)胞中起重要作用。近年來(lái)有不少文獻(xiàn)報(bào)道可溶性HLA-G二聚體能夠通過(guò)與抑制性受體ILT-4結(jié)合抑制DC的成熟，但也有人報(bào)道可溶性HLA-G二聚體對(duì)DC的成熟沒(méi)有影響。本研究擬探討可溶性HLA-G二聚體對(duì)DC成熟的作用，以期這種二聚體可以抑制DC的成熟，，進(jìn)一步誘導(dǎo)T細(xì)胞、B細(xì)胞的免疫耐受，降低移植排斥反應(yīng)的程度。巨噬細(xì)胞參與和促進(jìn)炎癥反應(yīng)，根據(jù)其微環(huán)境的不同可分為經(jīng)典的M1與選擇性激活的M2。前者與炎癥反應(yīng)和組織損傷有關(guān)；后者可促進(jìn)血管生成和組織修復(fù)，并且參與寄生蟲感染與過(guò)敏反應(yīng)，在一定條件下兩者可以相互轉(zhuǎn)化。由于巨噬細(xì)胞表面也存在可溶性HLA-G二聚體的受體，所以本實(shí)驗(yàn)將探討可溶性HLA-G二聚體對(duì)這兩種細(xì)胞極化的影響，研究可溶性HLA-G二聚體在炎癥反應(yīng)中的作用。論文分為2個(gè)部分，主要內(nèi)容與結(jié)果如下： 1.可溶性HLA-G二聚體在體外可抑制樹(shù)突狀細(xì)胞成熟 由人外周血分離得到PBMC，貼壁單核細(xì)胞在GM-CSF、IL-4的作用下分化成未成熟DC，而TNF-或IFN-γ可促進(jìn)DC成熟。同時(shí)在此過(guò)程中加入可溶性HLA-G二聚體，并設(shè)置721.221上清作為對(duì)照。用流式抗體CD1a，CD83，CD40染色，流式細(xì)胞儀檢測(cè)各組細(xì)胞表面這三種分子的表達(dá)水平。檢測(cè)結(jié)果顯示：可溶性HLA-G二聚體組DC表面CD83及CD40表達(dá)水平比721.221上清對(duì)照組明顯下降。 2.可溶性HLA-G二聚體在體外可抑制M1型巨噬細(xì)胞極化 由人外周血分離得到PBMC，貼壁單核細(xì)胞在GM-CSF與LPS刺激下得到M1；用M-CSF與IL-4刺激得到M2。并在實(shí)驗(yàn)組加入可溶性HLA-G二聚體，并設(shè)置721.221上清作為對(duì)照。用流式抗體CD14，CD80，CD206染色，流式細(xì)胞儀檢測(cè)M1組細(xì)胞表面CD14+CD80+的表達(dá)水平，M2組細(xì)胞表面CD14+CD206+的表達(dá)水平。流式細(xì)胞儀檢測(cè)結(jié)果顯示：可溶性HLA-G二聚體可以降低M1表面CD80的表達(dá)水平，而對(duì)M2表面的CD206表達(dá)沒(méi)有影響。
[Abstract]:HLA-G is a nonclassical HLA class I molecule that can induce extensive immune tolerance by binding to inhibitory receptors. A large number of studies have confirmed that soluble HLA-G dimer has stronger binding power with its inhibitory receptor and can induce stronger immune tolerance. These inhibitory receptors, including ILT-4 and ILT-2, are mainly expressed in T cell B cells, and macrophages, such as NKDC, are important antigen presenting cells linking innate and adaptive immunity, which can be found, captured, and captured. Exogenous antigen was processed and presented to T cells. Their interaction with primordial T cells plays an important role in inducing T cells to differentiate into Th1/Th2 cells and regulate T cells. In recent years, it has been reported that soluble HLA-G dimer can inhibit DC maturation by binding with inhibitory receptor ILT-4, but it is also reported that soluble HLA-G dimer has no effect on DC maturation. The aim of this study was to investigate the effect of soluble HLA-G dimer on DC maturation in order to inhibit DC maturation, induce T cell B cell immune tolerance and reduce the degree of graft rejection. Macrophages participate in and promote inflammatory response, according to their microenvironment can be divided into classic M1 and selectively activated M 2. The former is related to inflammation and tissue damage, while the latter can promote angiogenesis and tissue repair, and participate in parasitic infection and allergic reaction, which can transform each other under certain conditions. Because there are soluble HLA-G dimer receptors on the surface of macrophages, the effect of soluble HLA-G dimer on these two cell polarization and the role of soluble HLA-G dimer in inflammatory response were studied. The paper is divided into two parts. The main contents and results are as follows: 1. Soluble HLA-G dimer inhibits dendritic cell maturation in vitro PBMCs were isolated from human peripheral blood. Adherent monocytes differentiated into immature DCs under the action of GM-CSF- IL-4, while TNF- or IFN- 緯 promoted DC maturation. At the same time, soluble HLA-G dimer was added in the process, and 721.221 supernatant was set as control. Flow cytometry was used to detect the expression of these three molecules on the surface of the cells. The results showed that the expression of CD83 and CD40 on DC surface in soluble HLA-G dimer group was significantly lower than that in 721.221 supernatant group. 2. Soluble HLA-G dimer inhibits the polarization of M1 macrophages in vitro PBMCs were isolated from human peripheral blood. Monocytes were stimulated by GM-CSF and LPS to obtain M1 and M2 stimulated by M-CSF and IL-4. The soluble HLA-G dimer was added to the experimental group, and the supernatant of 721.221 was set as the control. Flow cytometry was used to detect the expression level of CD14 CD80 on the surface of M1 group and M2 group. The results of flow cytometry showed that soluble HLA-G dimer could decrease the expression of CD80 on M1 surface, but had no effect on CD206 expression on M2 surface.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 李丹;任亞娜;范華驊;;巨噬細(xì)胞的分類及其調(diào)節(jié)性功能的差異[J];生命科學(xué);2011年03期

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