發(fā)布時間：2018-01-13 02:13

  本文關(guān)鍵詞：分娩發(fā)動前后孕婦蛻膜及外周血中NK、NKT細(xì)胞生物學(xué)特性的研究　出處：《安徽醫(yī)科大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 自然殺傷T細(xì)胞 細(xì)胞因子 自然殺傷細(xì)胞 天然 分娩發(fā)動 自然殺傷細(xì)胞表面活化性受體D

【摘要】： 研究背景: 正常的妊娠,是母體免疫系統(tǒng)對帶有父源抗原的胎兒產(chǎn)生的一種免疫耐受現(xiàn)象,這種耐受是由多種因素之間相互制約、相互調(diào)節(jié)產(chǎn)生的。近年生殖免疫學(xué)者對自然殺傷細(xì)胞(NK cells)和自然殺傷T細(xì)胞(NKT cells)在其中的作用給予更多關(guān)注。NK細(xì)胞、NKT細(xì)胞在妊娠期產(chǎn)生Th2的免疫平衡,當(dāng)這種平衡被打破,即產(chǎn)生各種病理妊娠,并可能誘導(dǎo)產(chǎn)程的發(fā)動。人類分娩發(fā)動的機(jī)理至今仍是個迷,目前人們轉(zhuǎn)向組織水平、免疫水平找尋分娩動因。母-胎界面免疫微環(huán)境由蛻膜中的免疫活性細(xì)胞及其分泌的細(xì)胞因子組成,母體的免疫調(diào)節(jié)系統(tǒng)包括抗原特異T細(xì)胞、調(diào)節(jié)性T細(xì)胞、NK細(xì)胞、γδT細(xì)胞、NKT細(xì)胞和細(xì)CTL細(xì)胞等,它們共同參與調(diào)節(jié)Th1/Th2平衡構(gòu)成妊娠期Th2偏倚的免疫微環(huán)境,使母體對胎兒產(chǎn)生特異性免疫耐受以維持妊娠。 已有的研究證實,NK細(xì)胞大量存在于妊娠早期蛻膜并與胎盤絨毛滋養(yǎng)細(xì)胞密切接觸,在妊娠中發(fā)揮重要作用(其中70%左右為CD56brightCD16+NK細(xì)胞),而在妊娠晚期則明顯減少。最近大量的研究發(fā)現(xiàn),NKT細(xì)胞也參與了母胎耐受的形成。在人妊娠早期的蛻膜和外周血中存在NKT細(xì)胞的表達(dá),人蛻膜組織富集TCRαβ+CDl61+非恒定型NKT細(xì)胞(也稱為II型NKT細(xì)胞,non-iNKT cell)并表現(xiàn)了顯著的Th2型偏移(產(chǎn)生IL-4),為機(jī)體提供一個Th2型的微環(huán)境,并發(fā)現(xiàn)該群NKT細(xì)胞具有免疫抑制作用。提示蛻膜組織中的非恒定型NKT細(xì)胞可能通過提供一個Th2型的微環(huán)境從而抑制胎兒排斥和維持正常的妊娠。 本研究采用流式細(xì)胞熒光測定技術(shù)檢測孕足月分娩發(fā)動前后外周血和蛻膜組織單個核細(xì)胞中NK細(xì)胞、NKT細(xì)胞構(gòu)成及其表面NKG2D受體和NKG2A受體表達(dá)情況的變化,并進(jìn)一步檢測外周血單個核細(xì)胞中Th1/Th2平衡的變化及NK細(xì)胞、NKT細(xì)胞分泌Th1、Th2型細(xì)胞因子的情況,探討其與分娩發(fā)動的關(guān)系,揭示NK細(xì)胞及NKT細(xì)胞在分娩發(fā)動中的作用。并進(jìn)一步研究母胎界面趨化因子CXCR4的表達(dá)情況,試圖探討其與分娩發(fā)動前后NK細(xì)胞、NKT細(xì)胞重新分布之間的關(guān)系。研究外周血及蛻膜組織中NK細(xì)胞、NKT細(xì)胞的功能狀態(tài)與分娩發(fā)動的關(guān)系,無疑從一個全新的角度詮釋分娩這一生理現(xiàn)象,也為揭示妊娠時限異常疾病的發(fā)生機(jī)制及最終有效防治提供新思路,并為生殖免疫的發(fā)展提供新的理論依據(jù)。 目的: 檢測分娩發(fā)動前后蛻膜及外周血中NK細(xì)胞及NKT細(xì)胞的含量、表型、Th1/Th2型細(xì)胞因子表達(dá)方面的變化,探討NK細(xì)胞、NKT細(xì)胞獨特的表型、功能與分娩發(fā)動之間的重要聯(lián)系。檢測母胎界面趨化因子CXCR4表達(dá)的變化,以發(fā)現(xiàn)其與NK細(xì)胞、NKT細(xì)胞在產(chǎn)程發(fā)動后在母胎界面重新分布及功能變化的聯(lián)系。探討母胎免疫平衡在分娩發(fā)動中的作用。 方法: 1)收取正常晚孕分娩組(實驗組,32例)和正常晚孕剖宮產(chǎn)組(對照組,34例)壁蛻膜組織及同一人外周血,機(jī)械研磨法聯(lián)合密度梯度離心法分離出蛻膜單個核細(xì)胞,單純密度梯度離心法分離出外周血單個核細(xì)胞,流式細(xì)胞術(shù)檢測蛻膜及外周血中NK細(xì)胞、NKT細(xì)胞的含量及表型。 2)流式細(xì)胞術(shù)檢測正常晚孕分娩組和剖宮產(chǎn)組外周血中Th細(xì)胞平衡的變化及NK細(xì)胞、NKT細(xì)胞內(nèi)Th1/Th2型細(xì)胞因子IFN-γ和IL-4的表達(dá)。 3)收取正常晚孕自然分娩及剖宮產(chǎn)孕婦壁蛻膜及胎盤組織(各6例),RT-PCR方法檢測胎盤及壁蛻膜中CXCR4的表達(dá)情況。 結(jié)果: 1)與剖宮產(chǎn)組相比,分娩組蛻膜中NK細(xì)胞及NKT細(xì)胞的構(gòu)成低于剖宮產(chǎn)組,外周血中NK細(xì)胞及NKT細(xì)胞構(gòu)成高于剖宮產(chǎn)組(P0.01),兩組蛻膜中NK細(xì)胞構(gòu)成均高于外周血(P0.01)。 2)蛻膜單個核細(xì)胞中,分娩組CD56+NKG2A+細(xì)胞、CD56+ NKG2D+細(xì)胞、NK細(xì)胞表面NKG2A、NKG2D受體表達(dá)均低于剖宮產(chǎn)組(P0.05)。兩組中NKT細(xì)胞表面NKG2A、NKG2D受體表達(dá)無顯著差異(P0.05)。 3)外周血單個核細(xì)胞中,分娩組CD56+NKG2A+細(xì)胞構(gòu)成、NK細(xì)胞表面NKG2A受體表達(dá)低于剖宮產(chǎn)組,(P0.05)。CD56+NKG2D+細(xì)胞構(gòu)成、NK細(xì)胞表面NKG2D的表達(dá)及NKT細(xì)胞表面NKG2A/NKG2D受體的表達(dá)高于剖宮產(chǎn)組,(P0.01)。 4)外周血單個核細(xì)胞中,分娩組外周血CD3+T細(xì)胞中Th1細(xì)胞含量高于剖宮產(chǎn)組(P0.01),Th2細(xì)胞含量與剖宮產(chǎn)組無差異(P0.05)。分娩組NKT細(xì)胞、NK細(xì)胞分泌IFN-γ的能力高于剖宮產(chǎn)組。分娩組NKT細(xì)胞分泌IL-4的能力高于剖宮產(chǎn)組,NK細(xì)胞分泌IL-4的能力與剖宮產(chǎn)組無顯著差異。 5)兩組胎盤、胎膜均有CXCR4表達(dá),剖宮產(chǎn)組胎盤、胎膜上CXCR4表達(dá)相對高于分娩組。 結(jié)論: 分娩發(fā)動后外周血及蛻膜中NK細(xì)胞、NKT細(xì)胞的構(gòu)成、表型及功能有變化。分娩組蛻膜中NK細(xì)胞、NKT細(xì)胞含量降低,外周血中含量增加且傾向于Th1偏倚。外周血中NK細(xì)胞、NKT細(xì)胞中NKG2D受體表達(dá)增高,提示其功能活化與分娩發(fā)動有一定聯(lián)系,并且這種變化與趨化因子存在一定關(guān)聯(lián)。在分娩發(fā)動過程中免疫系統(tǒng)存在變化,不僅表現(xiàn)在全身,在母胎界面也有明顯變化,免疫平衡的改變可能在分娩發(fā)動中起著重要作用。
[Abstract]:Research background:
Normal pregnancy is a phenomenon of immune tolerance to maternal immune system with paternal antigens of the fetus, the tolerance is restrict each other by a variety of factors, the mutual regulation of reproductive immunology. In recent years, scholars on natural killer cells (NK cells) and natural killer T cells (NKT cells) to give more attention to.NK cells on the function of the immune balance of NKT cells to produce Th2 during pregnancy, when this balance is broken, which produce a variety of pathological pregnancy, and may start induction of labor. The mechanism of human parturition is still a mystery, the people turned to the organization level, the immune level of motivation. For delivery of maternal fetal immune micro the environment consists of immunocompetent cells in the decidua and the secretion of cytokines, including maternal immune system regulation of antigen specific T cells, regulatory T cells, NK cells, gamma delta T cells, NKT cells and small CTL Cells, etc., they participate in regulating Th1/Th2 balance to form immune microenvironment of Th2 bias during pregnancy, so that the mother has specific immune tolerance to the fetus to maintain pregnancy.
Research has confirmed that NK cells exist in early pregnancy decidua and trophoblast and placenta cells in close contact, play an important role in pregnancy (of which about 70% of CD56brightCD16+NK cells, and decreased) in late pregnancy. Many studies have found that NKT cells are involved in the formation of maternal fetal tolerance. Expression of NKT in cells during early pregnancy decidua and peripheral blood in human decidual tissue enriched TCR alpha beta +CDl61+ non constant type NKT cells (also known as II NKT cells, non-iNKT cell) and the performance of the Th2 type offset significantly (IL-4), provide a microenvironment for the Th2 body, and found the group of NKT cells with immunosuppression. Tips in decidua of non constant NKT cells may provide a microenvironment by Th2 to inhibit the fetal rejection and normal pregnancy.
This study used flow cytometry fluorescence detection of full-term delivery technology before and after the onset of NK cells in peripheral blood mononuclear cells and decidual tissues, changes of NKT cell structure and surface NKG2D receptor and NKG2A receptor expression, and further detected in peripheral blood mononuclear cells Th1 and NK cells balance changes of /Th2, NKT cells the secretion of Th1, Th2 type cytokines, to explore its relationship with the onset of labor, to reveal the role of NK cells and NKT cells in the initiation of labor. And further study on maternal fetal interface expression of chemokines CXCR4, to explore its and NK cells before and after parturition, the relationship between NKT cell NK cell redistribution. Study on peripheral blood and decidual tissues, the relationship between functional status and delivery of NKT cells to launch, no doubt the interpretation from a new perspective of delivery this physiological phenomenon, also to reveal the abnormal pregnancy time. The pathogenesis and the final effective prevention and control of disease provide new ideas, and provide new theoretical basis for the development of reproductive immunity.
Objective:
The phenotype of content detection before and after the onset of delivery in decidual and peripheral blood NK cells and NKT cells, the expression of Th1/Th2 type cytokines of NK cells, NKT cell phenotype unique and important link between the function and the onset of labor. The detection of maternal fetal interface of chemokines in the expression of CXCR4 and NK, to find the cells, the changes of NKT cells started in labor after redistribution in the maternal fetal interface and function. To investigate the role of maternal fetal immune balance in laboronset.
Method:
1) charged the normal fetal group (32 cases in experimental group, normal pregnant) and cesarean section group (control group, 34 cases) wall of decidua and peripheral blood of the same person, mechanical grinding method combined with density gradient centrifugation of decidual mononuclear cells isolated by density gradient centrifugation separation out peripheral blood mononuclear cells, NK cells were detected by flow cytometry in decidua and peripheral blood during the operation, the content and the phenotype of NKT cells.
2) flow cytometry was used to detect the changes of Th cell balance in peripheral blood of normal late pregnancy delivery group and cesarean section group, and the expression of Th1/Th2 cytokines IFN- gamma and IL-4 in NK cells and NKT cells.
3) the expression of CXCR4 in the placenta and the decidua of the placenta and wall decidua was detected by RT-PCR method for the normal delivery of late pregnancy and the caesarean section of the decidua and placenta (6 cases each).
Result:
1) compared with the cesarean section group, the NK cells and NKT cells in the decidua group were lower than those in the cesarean section group, and the NK cells and NKT cells in the peripheral blood were higher than those in the cesarean section group (P0.01). The NK cells in the two groups were higher than those in the peripheral blood (P0.01).
2) in decidual mononuclear cells, the expression of NKG2A and NKG2D receptors on CD56+NKG2A+ cells, CD56+ NKG2D+ cells and NK cells in the decidua mononuclear cells were lower than those in the cesarean section group (P0.05). There was no significant difference in the expression of NKG2A and NKG2D receptors on NKT cells in the two groups (P0.05).
3) in peripheral blood mononuclear cells, the CD56+NKG2A+ cells in the delivery group were made up. The expression of NKG2A receptors on the surface of NK cells was lower than that in the cesarean section group. (P0.05).CD56+NKG2D+ cells were formed, the expression of NKG2D on NK cells and the expression of NKG2A/NKG2D receptors on NKT cells were higher than those in caesarean section group (P0.01).
4) in peripheral blood mononuclear cells, Th1 cells in CD3+T cells in the peripheral blood of childbirth group is higher than that of cesarean section group (P0.01), Th2 cell content and cesarean section was no difference (P0.05). Delivery group NKT cells, NK cells secreted IFN- ability is higher than that of cesarean section group. Delivery capacity group NKT cells secreting IL-4 higher than that in the cesarean section group, NK cell secretory capacity of IL-4 and cesarean section group had no significant difference.
5) the expressions of placenta and placenta in the two groups were CXCR4, and the placenta in the caesarean section and the expression of CXCR4 on the membranes were higher than those in the delivery group.
Conclusion:
Parturition after NK cells in the peripheral blood and decidua, NKT cells, the phenotype and function change. NK cell delivery group in decidua of NKT cells decreased the content of content in peripheral blood increased and tended to bias Th1. NK cells in peripheral blood, increased expression of NKG2D receptor in NKT cells, suggesting that the the activation function and the onset of labor has certain relation, and this change and there is a certain correlation. Chemokines in laboronset immune system in the process of change, not only in the body, at the maternal fetal interface also has obvious changes, the change of the balance of immunity may play an important role in the onset of labor.

【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 ;Cell-Mediated Immunity Imbalance in Patients with Intrahepatic Cholestasis of Pregnancy[J];Cellular & Molecular Immunology;2007年01期

2 ;Interferon-γ Expression in Natural Killer Cells and Natural Killer T Cells Is Suppressed in Early Pregnancy[J];Cellular & Molecular Immunology;2007年05期

3 龔衛(wèi)娟;KIR研究進(jìn)展[J];國外醫(yī)學(xué)(免疫學(xué)分冊);2003年02期

4 王軼英;;正常妊娠不同孕期胎盤組織中CXCR4及SDF1的表達(dá)[J];河南大學(xué)學(xué)報(醫(yī)學(xué)版);2008年01期

5 李莉平;林羿;康佳麗;夏薇;王自能;;早、中、晚孕期胎盤因子對人外周血淋巴細(xì)胞CD4、CCR5和CXCR4表達(dá)的影響[J];免疫學(xué)雜志;2007年04期

6 沈亞娟;張建;田志剛;;腫瘤患者自然殺傷細(xì)胞受體NKG2A和NKG2D的表達(dá)[J];檢驗醫(yī)學(xué);2008年04期

7 劉景華;于力;;NKT細(xì)胞研究進(jìn)展[J];細(xì)胞與分子免疫學(xué)雜志;2008年02期

8 祝愛霞;鄒建話;蔡葉琴;何水群;馬鳳蘭;;健康足月分娩母親與新生兒T細(xì)胞亞群和NK細(xì)胞水平[J];中國婦幼保健;2007年22期

9 熊苗;屠菊紅;趙愛民;;母胎界面趨化因子及其受體的研究進(jìn)展[J];中國婦幼保健;2007年23期

10 陳自勵;;早產(chǎn)兒主要并發(fā)癥防治進(jìn)展[J];中國實用婦科與產(chǎn)科雜志;2008年05期

，

本文編號：1416994