本文選題：IL-12 + IL-12受體��； 參考：《濱州醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：探索樹(shù)突狀細(xì)胞(DC)與蛻膜自然殺傷細(xì)胞(uNK)之間的相互作用在剛地弓形蟲(chóng)(Toxoplasma gondii)感染致不良妊娠結(jié)局中分子免疫機(jī)制。方法：體外純化的來(lái)自正常妊娠的C57BL/6小鼠的uNK細(xì)胞和正常小鼠骨髓誘導(dǎo)而來(lái)的DCs以5：1的比例在弓形蟲(chóng)感染或不感染的情況下進(jìn)行共培養(yǎng)24小時(shí)。設(shè)transwell小室隔開(kāi)組和直接接觸組。同時(shí)建立弓形蟲(chóng)感染C57BL/6孕鼠模型。體內(nèi)將24只C57BL/6孕鼠按完全隨機(jī)分組法分為感染組(12只)和對(duì)照組(12只),感染組小鼠于孕8天每只腹腔注射400個(gè)RH株剛地弓形蟲(chóng)速殖子,對(duì)照組注射等量無(wú)菌PBS,所有孕鼠均于孕14天時(shí)處死,觀察胎盤(pán)及胎鼠的發(fā)育情況。用流式細(xì)胞儀分析uNK細(xì)胞上白細(xì)胞介素12受體(IL-12R)、抑制性受體NKG2A和活化性受體NKG2D的表達(dá)情況。用ELISA方法測(cè)定細(xì)胞培養(yǎng)上清和胎盤(pán)中IL-12和IFN-y的水平。共培養(yǎng)組并加設(shè)抗IL-12抗體組。用流式細(xì)胞儀檢測(cè)uNK細(xì)胞的殺傷活性。結(jié)果：體內(nèi)實(shí)驗(yàn),在孕14d感染組小鼠的胎盤(pán)出血壞死較多,未感染的對(duì)照組小鼠胎盤(pán)血供豐富。感染組小鼠的死胎率比對(duì)照組小鼠顯著增加(p0.01)。弓形蟲(chóng)感染后IL-12R、NKG2A和NKG2D的表達(dá)水平均上調(diào),但活化性受體NKG2D比抑制性受體NKG2A升高更為明顯。共培養(yǎng)組在弓形蟲(chóng)感染后NKG2D的表達(dá)水平及uNK細(xì)胞的細(xì)胞毒性比單獨(dú)uNK細(xì)胞感染組升高明顯,且共培養(yǎng)組上清液中的IL-12和IFN-y水平比單獨(dú)的uNK細(xì)胞組、單純DC組升高明顯。在共培養(yǎng)組加入抗IL-12抗體時(shí),IFN-γ的增強(qiáng)不明顯。在感染孕鼠的胎盤(pán)中IL-12和IFN-y的水平較正常孕鼠組也顯著增加。結(jié)論：弓形蟲(chóng)感染后,DC細(xì)胞通過(guò)分泌IL-12與uNK細(xì)胞表面的IL-12R相互作用,從而促進(jìn)uNK細(xì)胞的活化、IFN-γ分泌增多和細(xì)胞毒性增強(qiáng)從而導(dǎo)致不良妊娠結(jié)局的發(fā)生,這可能是弓形蟲(chóng)感染致不良妊娠結(jié)局的重要的免疫機(jī)制之一。
[Abstract]:Aim: to explore the molecular immune mechanism of the interaction between dendritic cell (DC) and decidua natural killer (NK) in the adverse pregnancy outcome caused by Toxoplasma gondii infection (Toxoplasma gondii). Methods: purified uNK cells from normal pregnant C57BL/6 mice and DCs from bone marrow of normal mice were cultured for 24 hours with or without Toxoplasma gondii infection at 5:1. Transwell compartment group and direct contact group were established. At the same time, the pregnant rat model of C57BL/6 infection with Toxoplasma gondii was established. Twenty-four pregnant C57BL/6 mice were randomly divided into infected group (n = 12) and control group (n = 12). Four hundred Toxoplasma gondii Tachyzoites of RH strain were injected intraperitoneally in infected mice on the 8th day of gestation. All the pregnant rats were killed on the 14th day of gestation. The development of placenta and fetal mice were observed. The expression of interleukin-12 receptor (IL-12), inhibitory receptor (NKG2A) and activated receptor (NKG2D) in uNK cells were analyzed by flow cytometry. The levels of IL-12 and IFN-y in cell culture supernatant and placenta were measured by ELISA method. Co-culture group and anti-IL-12 antibody group were added. The cytotoxicity of uNK cells was detected by flow cytometry. Results: in vivo experiment, the placenta hemorrhage and necrosis were more in the infected mice on the 14th day of gestation, and the placental blood supply was abundant in the uninfected control group. The stillbirth rate of infected mice was significantly higher than that of control group (P 0.01). After Toxoplasma gondii infection, the expression of IL-12 RN NKG2A and NKG2D were up-regulated, but the activated receptor NKG2D was significantly higher than the inhibitory receptor NKG2A. The expression of NKG2D and the cytotoxicity of uNK cells in co-culture group were significantly higher than those in uNK cell group, and the levels of IL-12 and IFN-y in supernatant of co-culture group were significantly higher than those in uNK cell group and DC group. The increase of IFN- 緯 was not obvious when anti-IL-12 antibody was added in co-culture group. The levels of IL-12 and IFN-y in the placenta of infected pregnant mice were significantly higher than those of normal pregnant mice. Conclusion: after Toxoplasma gondii infection, DC cells secrete IL-12 and interact with IL-12R on the surface of uNK cells, thus promoting the activation of uNK cells to increase the secretion of IFN- 緯 and the enhancement of cytotoxicity, resulting in the occurrence of adverse pregnancy outcomes. This may be an important immune mechanism for adverse pregnancy outcomes caused by Toxoplasma gondii infection.
【學(xué)位授予單位】：濱州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R714.251
，

本文編號(hào)：1838516