本文選題：瘧疾 + 調(diào)節(jié)性B細(xì)胞��； 參考：《中國科學(xué)技術(shù)大學(xué)》2013年博士論文

【摘要】：瘧疾是由原蟲類寄生蟲——瘧原蟲(Plasmodium spp.)感染引起的流行性疾病,是全球三大主要傳染性疾病之一。流行病學(xué)調(diào)查數(shù)據(jù)表明,目前在世界許多地區(qū),瘧疾仍然呈現(xiàn)高發(fā)病率和高死亡率,特別是在撒哈拉以南的非洲地區(qū)；易感人群多為5歲以下的兒童,其感染后常表現(xiàn)嚴(yán)重的病理變化和癥狀,例如腦瘧、嚴(yán)重貧血和肺功能衰竭等。然而,我們對瘧疾感染所誘導(dǎo)的主要免疫病理變化—腦瘧—及其調(diào)控機(jī)理尚未完全了解。在瘧疾流行區(qū),感染瘧疾后的人群雖然能獲得部分保護(hù)性免疫,但是這種免疫力維持時間短,并且在個體離開瘧疾流行區(qū)后保護(hù)力快速消失。目前治療瘧疾的主要手段是藥物治療,可是由于瘧原蟲耐藥性的產(chǎn)生及藥物的不及時使用嚴(yán)重降低抗瘧藥的效果,所以疫苗被認(rèn)為是目前應(yīng)對瘧疾的最理想方式。然而在瘧疾疫苗研發(fā)方面我們?nèi)匀幻媾R巨大的挑戰(zhàn),其原因之一是我們尚未完全探明瘧原蟲病原與宿主之間的復(fù)雜免疫學(xué)關(guān)系。在本論文報告的研究中,我們采用小鼠伯氏瘧原蟲(Plasmodium berghei)瘧疾模型,(1)分析研究了與瘧疾相關(guān)的主要免疫病理變化—腦瘧的發(fā)生和調(diào)節(jié)機(jī)制；(2)比較和分析了誘導(dǎo)有效免疫保護(hù)力的瘧疾疫苗的不同免疫策略。 1.調(diào)節(jié)性B細(xì)胞與腦瘧 瘧原蟲感染誘發(fā)的神經(jīng)系統(tǒng)功能障礙—腦瘧,是瘧疾感染時產(chǎn)生的主要病理變化和致死原因之一。大約有1%的惡性瘧原蟲感染個體可發(fā)生腦瘧；其中,10%到20%的腦瘧患者發(fā)生死亡。腦瘧是免疫失調(diào)所導(dǎo)致的腦組織免疫病理反應(yīng)。目前認(rèn)為,其主要的誘發(fā)原因與感染瘧原蟲的紅細(xì)胞粘附在腦組織毛細(xì)血管壁,并引起局部毛細(xì)血管堵塞和炎性細(xì)胞招募有關(guān)。淋巴細(xì)胞浸潤、免疫殺傷性NK細(xì)胞和細(xì)胞毒性CD8+T細(xì)胞等破壞毛細(xì)血管壁和血腦屏障,導(dǎo)致腦組織出血和神經(jīng)系統(tǒng)功能受損。 免疫調(diào)節(jié)性細(xì)胞因子IL-10在維持免疫自身穩(wěn)定和控制感染誘導(dǎo)的免疫病理如腦瘧的過程中發(fā)揮關(guān)鍵作用。近年來有大量實驗表明,可分泌IL-10的調(diào)節(jié)性B細(xì)胞具有免疫調(diào)節(jié)功能,并證實其在維持免疫自身耐受和抑制免疫病理中發(fā)揮著重要作用。據(jù)此,本課題采用伯氏瘧原蟲感染C57BL/6小鼠的瘧疾模型,研究了調(diào)節(jié)性B細(xì)胞在瘧疾感染過程中調(diào)控過度免疫反應(yīng)和腦瘧病理的作用。 研究結(jié)果表明,伯氏瘧原蟲感染后可誘導(dǎo)血清中IL-10水平的增高；同時觀察到小鼠脾臟細(xì)胞中可分泌IL-10的調(diào)節(jié)性B細(xì)胞比率顯著增高。體內(nèi)轉(zhuǎn)輸實驗證實,IL-10+調(diào)節(jié)性B細(xì)胞明顯抑制腦組織微血管出血性病理變化,并顯著降低瘧疾感染小鼠的死亡率；進(jìn)一步研究發(fā)現(xiàn),調(diào)節(jié)性B細(xì)胞能抑制免疫殺傷細(xì)胞向腦組織遷移,但對瘧原蟲血蟲水平?jīng)]有顯著影響。用IL-10受體抗體阻斷IL-10信號后,調(diào)節(jié)性B細(xì)胞抑制腦瘧發(fā)生的作用消失,這說明調(diào)節(jié)性B細(xì)胞抑制腦瘧發(fā)生是由IL-10介導(dǎo)的。我們還發(fā)現(xiàn),轉(zhuǎn)輸CD4+CD25+調(diào)節(jié)性T細(xì)胞對腦瘧發(fā)生沒有顯著作用。以上結(jié)果表明調(diào)節(jié)性B細(xì)胞,而不是調(diào)節(jié)性T細(xì)胞,抑制瘧原蟲感染的小鼠腦瘧病理的發(fā)生。此項研究的成果不僅揭示瘧疾病理新的調(diào)控機(jī)制,還對制定腦瘧預(yù)防策略及抗瘧疾藥物研發(fā)提供重要理論依據(jù)。 2.瘧疾活蟲疫苗的免疫效能 目前疫苗被認(rèn)為是控制瘧疾最有效的方式。近半個多世紀(jì)以來人們嘗試用各種疫苗方式來預(yù)防瘧疾,然而到目前為止還沒有成功的抗瘧疾疫苗在臨床上應(yīng)用。亞單位疫苗被人們寄予厚望,但近三十年來許多亞單位瘧疾疫苗臨床試驗都以失敗告終,這可能與這些疫苗缺乏危險信號進(jìn)而不能有效激活免疫應(yīng)答和免疫保護(hù)力相關(guān)。人們發(fā)現(xiàn)放射線致弱和基因改造的減毒子孢子能夠誘導(dǎo)針對瘧疾的完全保護(hù)力,可是這種疫苗難以在體外大規(guī)模獲得從而限制了在臨床上的應(yīng)用。 在本研究中,我們分析兩種疫苗免疫策略誘導(dǎo)的免疫應(yīng)答和保護(hù)力水平。一種是短期活蟲感染加藥物終止模擬活蟲疫苗的方式,另一種是全蟲抗原加佐劑的疫苗方式。研究目的是鑒定兩種疫苗策略的免疫應(yīng)答差異以及與有效保護(hù)力相關(guān)的免疫應(yīng)答標(biāo)志物。實驗結(jié)果表明,短期活蟲感染免疫可誘導(dǎo)較強(qiáng)的免疫保護(hù)力。二全蟲抗原家佐劑僅可誘導(dǎo)部分保護(hù)。與全蟲抗原加佐劑組相比,短期活蟲感染加藥物阻斷組誘導(dǎo)更高的瘧原蟲特異的抗體水平和樹突狀細(xì)胞激活水平以及更強(qiáng)的CD4+T細(xì)胞應(yīng)答。Thl型、Th2型和調(diào)節(jié)性細(xì)胞因子表達(dá)水平在兩種疫苗策略之間都有類似的顯著性差異。進(jìn)一步的研究將驗證這些免疫因素和保護(hù)力之間的相互聯(lián)系。本項研究有助于設(shè)計出更理性的瘧疾疫苗方案。
[Abstract]:Malaria is a protozoan parasite Plasmodium (Plasmodium spp.) infection caused by epidemic disease, is one of the three major infectious diseases worldwide. Epidemiological data show that in many parts of the world, malaria still has high morbidity and mortality, especially in sub Saharan Africa; the susceptible population is children under the age of 5, after its infection often show pathological changes and serious symptoms, such as cerebral malaria, severe anemia and lung failure. However, we change to the main immune pathology of malaria infection induced by cerebral malaria and its regulation mechanism is not yet fully understood. In malaria endemic areas, malaria infection after the crowd although we can obtain some protective immunity, but the immunity of short duration, and in individual leaves in malaria endemic area after the protection force quickly disappeared. The main therapy for the treatment of malaria The drug treatment, but because of the drug resistance of Plasmodium and not using seriously reduce the antimalarial effect, so the vaccine is considered to be the ideal way to deal with malaria at present. However, in the aspect of malaria vaccine development we still face enormous challenges, one of the reasons is that we have not yet fully proved the complex immunological relationship between Plasmodium the pathogen and host. In this research report, we used mouse Plasmodium berghei malaria model (Plasmodium berghei), (1) analysis of the occurrence of cerebral malaria changes mainly immunopathology associated with malaria and regulatory mechanisms; (2) comparison and analysis of different immunization strategies effective malaria vaccine induced immune protection force.
1. regulatory B cells and brain malaria
The dysfunction of brain nervous system induced by malaria Plasmodium infection, is the main pathological changes in malaria infection and cause of death. The occurrence of cerebral malaria infected individuals about 1% of Plasmodium falciparum; among them, 10% to 20% of the deaths occurred in patients with cerebral malaria. Malaria is the brain pathological immune response immune disorders caused by brain tissue. At present, red cell adhesion and its main causes and infection of Plasmodium in brain capillary wall, and cause local capillary congestion and inflammatory cell recruitment. Lymphocyte infiltration, immune killer NK cells and cytotoxic CD8+T cells destroy the capillary wall and the blood-brain barrier, leading to bleeding and neurological damage of brain.
Immunoregulatory cytokine IL-10 play a key role in the process of maintaining immune homeostasis and immune control of infection induced such as cerebral malaria. In recent years a large number of experiments show that the secretion of IL-10 regulatory B cells have immune regulatory function, and confirmed in maintaining immunological self tolerance and plays an important role in inhibiting immune pathology in this paper. Accordingly, the malaria model C57BL/6 mice infected with Plasmodium berghei, studied the regulatory B cells control excessive immune response and pathological cerebral malaria in malaria infection process.
The results show that the increase of Plasmodium berghei infection can induce IL-10 level in serum; at the same time observed in mouse spleen cells can secrete IL-10 regulatory B cell ratio was significantly increased. In vivo transfer experiment confirmed that IL-10+ regulatory B cells inhibit hemorrhagic cerebral microvascular pathological changes, and significantly reduce malaria infection the death rate of mice; further study found that regulatory B cells can inhibit the immune cell migration into the brain, but had no significant effect on parasite blood worms level. Blocking IL-10 signal with IL-10 receptor antibodies after disappearance of regulatory B cells inhibit cerebral malaria occurred, suggesting that regulatory B cells inhibit cerebral malaria occurs mediated by IL-10. We also found that the transfer of CD4+CD25+ regulatory T cells on cerebral malaria has no significant effect. The above results show that regulatory B cells, rather than inhibition of regulatory T cells. The occurrence of cerebral malaria in mice infected with Plasmodium falciparum can not only reveal the new regulation mechanism of malaria pathology, but also provide important theoretical basis for formulating strategies for preventing cerebral malaria and developing anti malaria drugs.
2. immune efficacy of a live worm vaccine
The vaccine is considered to be the most effective way to control malaria. For more than half a century of people trying to use various methods to prevent malaria vaccine, but so far there is no anti malaria vaccine successfully applied in clinic. The subunit vaccine were high hopes, but in the past thirty years many subunit malaria vaccine clinical trials ended in failure, which may be related to the lack of effective vaccine danger signals and can not activate the immune response and protective immunity. People find complete protection of radiation induced reduction of sub toxic spores and weak genetic modification can induce the vaccine against malaria, but difficult to obtain large scale in vitro which limits the clinical application.
In this study, we analyzed two kinds of strategies of vaccine induced immune immune response and protection level. One is to terminate the simulation of living vaccine and drug short live insect infection, the other is a vaccine plus whole worm antigen adjuvant. The purpose of the study is to identify two kinds of differences in immune response and immune vaccine strategies the response associated with the effective protection of the marker. The experimental results show that the short live insects infection can induce strong immune protection. Two whole antigens can only be induced home adjuvant protection. Compared with the whole antigens plus adjuvant group, short live insects infected with drugs blocking antibody level and dendritic cells to induce specific higher group the activation level of Plasmodium and stronger CD4+T cell response to.Thl, the expression level of Th2 and regulatory cytokines have significant differences between the two kinds of similar vaccine strategy further. The study will verify the interlink between these immune factors and protective power. This study helps to design a more rational scheme for malaria vaccine.

【學(xué)位授予單位】：中國科學(xué)技術(shù)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R531.3

【共引文獻(xiàn)】

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相關(guān)碩士學(xué)位論文 前10條

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