本文選題：免疫耐受 + 包蟲病　； 參考：《新疆醫(yī)科大學(xué)》2016年博士論文

【摘要】：目的:包蟲病是全球人畜共患性疾病。包蟲病所致的過敏性休克是其嚴(yán)重并發(fā)癥,占包蟲病患者突然死亡原因的20%。包蟲病患者存在免疫耐受已被證實,但是這種狀態(tài)在包蟲囊液外溢時發(fā)生轉(zhuǎn)變,過敏性休克發(fā)生,具體機制尚待闡明。為此,本研究在完善囊型包蟲病所致過敏反應(yīng)動物模型、優(yōu)化小鼠過敏反應(yīng)評價體系的基礎(chǔ)上,探討以Treg細(xì)胞為核心的“免疫耐受調(diào)節(jié)網(wǎng)絡(luò)”中相關(guān)重要因子在囊型包蟲病所致過敏反應(yīng)中的變化及特點,以及地塞米松預(yù)處理對囊型包蟲病所致過敏反應(yīng)的影響及機制,為制定針對囊型包蟲病所致過敏反應(yīng)的有效防治策略奠定理論基礎(chǔ)。方法:1)羊源細(xì)粒棘球蚴原頭節(jié)及囊液均采自包蟲感染羊肝臟。包蟲感染組,每只小鼠通過腹腔接種粒棘球蚴原頭節(jié)2000個,建立囊型包蟲感染模型,再應(yīng)用羊源細(xì)粒棘球蚴粗制囊液0.1ml/10g攻擊發(fā)敏建立囊型包蟲病所致過敏反應(yīng)模型。通過觀察過敏癥狀、肛溫變化、肺組織HE染色病理切片,測定肺泡灌洗液中白細(xì)胞分類計數(shù)、過敏介質(zhì)組胺及PAF-AH水平,測量血清抗體水平等一系列指標(biāo),全面優(yōu)化過敏評價體系,完善過敏模型建立;2)BALB/c小鼠隨機分成健康對照組、包蟲感染組、囊液致敏組。使用全面的過敏評價體系評價過敏模型。FCM檢測小鼠淋巴結(jié)來源PBMCs中DCs、Treg細(xì)胞水平,ELISA檢測血清IgE、IgG、IgG1、IL-10、TGF-β1、IL-13、IL-17A水平;3)C57BL/6小鼠隨機分成健康對照組、單純囊液致敏組,地塞米松預(yù)處理組。在包蟲囊液致敏前30min腹腔注射地塞米松建立地塞米松預(yù)處理模型。使用全面的過敏評價體系評價過敏模型。FCM檢測小鼠淋巴結(jié)來源PBMCs中Treg細(xì)胞水平,ELISA檢測血清IgE、IgG、IgG1、IL-10、TGF-β1、IL-13水平。結(jié)果:1)BALB/c小鼠包蟲接種成功率為75%,C57BL/6小鼠包蟲接種成功率為65%,囊型包蟲病所致過敏模型成功率為分別為93.33%和85.62%。發(fā)生過敏反應(yīng)小鼠出現(xiàn)了不同程度的過敏癥狀、肛溫下降、肺組織HE病理切片及BALF中炎性細(xì)胞增加、組胺表達量增加、血清抗體IgE、IgG、IgG1水平增加。上述指標(biāo)均表明BALB/c和C57BL/6小鼠囊型包蟲病所致過敏反應(yīng)模型建立成功;2)與健康對照組相比,包蟲感染組及囊液致敏組小鼠血清IgE水平均明顯增加(P0.01)。囊液致敏組小鼠血清IgE水平明顯高于包蟲感染組(P0.05)。囊液致敏后,肺組織病理及BALF中炎性細(xì)胞增加、組胺表達水平增加、血清IL-13、IL-17A水平增加(P0.05)。與健康對照組相比,小鼠血清IL-10、TGF-β1水平及CD4+CD25+Foxp3+Treg/CD4+細(xì)胞比例在包蟲感染組增加,在囊液致敏組下降(P0.001),呈現(xiàn)先升后降的雙向變化。但是其他T細(xì)胞群,如CD4+CD25+Foxp3-細(xì)胞比例、CD4+Foxp3+CD25-細(xì)胞比例,各組間比較無明顯差異。CD11c+IA/IE+CD86+細(xì)胞比例、CD11c+IA/IE+CD80+細(xì)胞比例以及CD4/CD8比例,各組間比較無明顯差異;3)小鼠嚴(yán)重過敏反應(yīng)發(fā)生率在地塞米松預(yù)處理組是12.5%,單純囊液致敏組是37.5%。與單純囊液致敏組相比,地塞米松預(yù)處理組小鼠肺組織及BALF中炎性細(xì)胞降低、組胺表達水平減少、IgE水平降低、血清IL-13水平降低(P0.001)。血清IL-10及TGF-β1水平與CD4+CD25+Foxp3+Treg/CD4+細(xì)胞比例變化趨勢相同,均在單純囊液致敏組下降,在地塞米松預(yù)處理組增加。結(jié)論:1)全面的小鼠過敏評價體系優(yōu)化了囊型包蟲病所致過敏反應(yīng)小鼠動物模型,為探討包蟲病所致過敏反應(yīng)中免疫耐受機制奠定基礎(chǔ);2)以Treg細(xì)胞為核心的“免疫耐受調(diào)節(jié)網(wǎng)絡(luò)”維持了包蟲感染后的免疫耐受狀態(tài),細(xì)粒棘球蚴囊液致敏后,網(wǎng)絡(luò)失衡,誘發(fā)過敏反應(yīng);3)包蟲感染后以Treg細(xì)胞為核心的“免疫耐受調(diào)節(jié)網(wǎng)絡(luò)”通過CD25、Foxp3高表達,IL-10、TGF-β1高分泌水平,維持免疫耐受功能。囊液致敏后,免疫耐受狀態(tài)發(fā)生轉(zhuǎn)變,Treg細(xì)胞比例下降,IL-10、TGF-β1水平降低,IL-17A、IL-13水平升高,發(fā)生過敏反應(yīng);4)在包蟲病所致的過敏反應(yīng)中,地塞米松可通過降低IgE水平,減少肺組織及BALF中炎性細(xì)胞滲出,減少靶器官過敏介質(zhì)組胺釋放,減輕過敏反應(yīng);5)地塞米松通過上調(diào)Treg細(xì)胞比例及功能維持“免疫耐受調(diào)節(jié)網(wǎng)絡(luò)”功能。其具體機制是上調(diào)IL-10和TGF-β1水平,抑制Th2細(xì)胞因子IL-13水平。
[Abstract]:Objective: hydatid disease is a global zoonotic disease. Hydatid disease caused by anaphylactic shock is the serious complications, accounted for 20%. of hydatid disease hydatid disease suddenly causes of death in patients with the presence of immune tolerance has been confirmed, but this change in hydatid fluid overflow, allergic shock, tillneeds to clarify. In this study, due to cystic echinococcosis anaphylaxis animal model improvement, optimization based on the evaluation system of anaphylaxis in mice, to investigate the Treg cells as the core of the "immune tolerance regulation network related important factor in cystic hydatid disease caused by allergic reactions in changes and characteristics, and the effect of dexamethasone pretreatment on cystic echinococcosis disease The impact caused by the allergic reaction and mechanism, which lays the theoretical foundation for the formulation of effective prevention strategies for cystic hydatid disease caused by an allergic reaction. Methods: 1) sheep derived Echinococcus granulosus protoscolex and cyst fluid were collected from sheep liver hydatid infection. Infection group, each mouse by intraperitoneal inoculation of grain granulosus protoscolex 2000 and the establishment of cystic echinococcosis Infection model using sheep derived Echinococcus granulosus cystic fluid 0.1ml/10g attack caused by a sensitive cystic echinococcosis allergic reaction model. Through the observation of allergic symptoms, changes of rectal temperature, HE staining of lung tissue pathological sections in bronchoalveolar lavage fluid in white blood cell count, allergic medium histamine and PAF-AH levels, the measurement of serum antibody level etc. A series of indexes, comprehensive optimization of allergy evaluation system, improve the allergy model; 2) BALB/c mice were randomly divided into control group, infection group, allergy group. The cystic fluid evaluation system of comprehensive evaluation of the use of allergy allergy model.FCM mice were detected from lymph nodes in PBMCs DCs and Treg cell levels, serum IgE, ELISA, IgG IgG1, IL-10, TGF-, beta 1, IL-13, IL-17A; 3) C57BL/6 mice were randomly divided into healthy control group, simple cystic fluid sensitized group, dexamethasone pretreatment group. In hydatid fluid before sensitization by intraperitoneal injection of 30min dexamethasone to establish dexamethasone pretreatment model. Using the evaluation system of comprehensive evaluation of allergic allergy detection model.FCM mouse lymph node cell Treg PBMCs in The level of serum IgE, ELISA, IgG, IgG1, IL-10, TGF- beta 1, IL-13 1). Results: the success rate of BALB/c mice inoculated with echinococcosis was 75%, the success rate of C57BL/6 mice inoculated with 65% hydatid cystic hydatid disease caused by allergy, the success rate of the model was respectively 93.33% and 85.62%. mice developed allergic allergic reaction the symptoms of different degree, rectal temperature decreased, The inflammatory cells of HE pathology and BALF in lung tissue increased, the increase in expression of histamine, serum antibody IgE, IgG, IgG1. The above indexes showed increased levels of BALB/c and C57BL/6 in mice caused by cystic echinococcosis allergic reaction model; 2) compared with healthy control group, infection group and hydatid cyst fluid sensitized group serum IgE levels were significantly increased Plus (P0.01). The cystic fluid sensitized mice serum IgE levels were significantly higher than that of hydatid infection group (P0.05). Sensitized cystic fluid, lung tissue pathology and BALF in inflammatory cells increased, the expression level of histamine increased, serum IL-13, IL-17A levels increased (P0.05). Compared with the healthy control group, serum IL-10 rat, TGF- beta 1 and CD4+CD25+Foxp3+Treg/CD4+ cell ratio In cases of hydatid infection group, sensitized group in cyst fluid decreased (P0.001), shows the first increased and then decreased. But the other T cells, such as CD4+CD25+Foxp3- cell ratio, CD4+Foxp3+CD25- cell ratio,.CD11c+IA/IE+CD86+ cell ratio is no significant difference between groups, the proportion of CD11c+IA/IE+CD80+ cells and the ratio of CD4/CD8, ratio between groups There are no significant difference; 3) severe allergic reactions in mice in dexamethasone pretreatment group is 12.5%, the Dan Chunnang solution sensitized group is 37.5%. with simple cystic fluid sensitized group, dexamethasone pretreatment of inflammatory cells in lung tissues of mice in BALF and decreased the expression level of histamine decreased, the decrease of IgE level, the level of serum IL-13 lower serum I (P0.001). L-10 and TGF- beta 1 level and the percentage of CD4+CD25+Foxp3+Treg/CD4+ cells in the same trend, both in pure cystic fluid sensitized group decreased, increased in the dexamethasone treated group. Conclusion: 1) a comprehensive evaluation system to optimize the mouse allergic reaction caused by cystic hydatid disease in mice animal model, to explore the immune induced allergic reaction in hydatid disease resistance By laying the foundation mechanism; 2) to Treg cells as the core of the "tolerance regulation network" to maintain the infection after immune tolerance, sensitization of Echinococcus granulosus cyst fluid, network imbalance, cause an allergic reaction; 3) hydatid after infection with Treg cells as the core of the immune tolerance regulation network by CD25. The high expression of Foxp3, IL-10, TGF- beta 1 High level, maintain tolerance function. The cysts after sensitization, immune tolerance state changes, the proportion of Treg cells decreased IL-10, TGF- beta 1 reduced the level of IL-17A, IL-13 increased the level of allergic reaction; 4) in hydatid disease caused by allergic reaction, dexamethasone can reduce IgE level, reduce inflammatory cell infiltration in lung tissue and BALF Out, reduce target organ allergy mediators histamine release, reduce allergic reactions; 5) dexamethasone through upregulation of Treg cell proportion and maintain the function of "immune tolerance regulation network function. Its mechanism is upregulated IL-10 and TGF- beta 1 level, the level of Th2 cell inhibitory factor IL-13.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R532.32
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本文編號：2047935