本文選題：轉(zhuǎn)化生長因子β　切入點(diǎn)：白介素2　出處：《中國科學(xué)技術(shù)大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：第一部分細(xì)胞因子對(duì)調(diào)節(jié)性T細(xì)胞的發(fā)育調(diào)節(jié)研究揭示誘導(dǎo)機(jī)體免疫耐受打破的因素對(duì)于理解自身免疫性疾病的發(fā)生發(fā)展有重要作用。Treg細(xì)胞是控制機(jī)體過度免疫應(yīng)答阻止自身免疫性疾病發(fā)生的負(fù)調(diào)性細(xì)胞,越來越多的研究也表明Treg還參與到細(xì)菌及病毒感染、腫瘤、器官移植耐受、組織修復(fù)等多種疾病過程。因此,對(duì)Treg發(fā)育調(diào)節(jié)的研究有利于進(jìn)一步認(rèn)識(shí)Treg的功能,從而將其作為多種疾病的治療靶點(diǎn)。Treg細(xì)胞可由胸腺發(fā)育及外周誘導(dǎo)兩種途徑產(chǎn)生,其發(fā)育分化受到細(xì)胞因子的調(diào)節(jié)。在CD4啟動(dòng)子下表達(dá)顯性失活TGF-β⒏Ⅱ型受體的dnTGF-βRⅡ小鼠會(huì)出現(xiàn)T細(xì)胞過度活化而導(dǎo)致的肝臟和腸道炎性細(xì)胞浸潤,其Treg細(xì)胞的抑制功能降低。Il2ra-/-小鼠由于Treg功能失調(diào),出現(xiàn)多克隆T細(xì)胞和B細(xì)胞的擴(kuò)增而導(dǎo)致多臟器自身免疫性疾病。在本研究中,我們利用Il2ra-/-Tg小鼠,探究TGF-β與IL-2信號(hào)在Treg發(fā)育過程中的共同調(diào)節(jié)作用。我們發(fā)現(xiàn)Il2ra-/-Tg小鼠出現(xiàn)類似于Treg缺陷的scurfy小鼠癥狀,多個(gè)臟器出現(xiàn)淋巴細(xì)胞浸潤與病理損傷。Il2ra-/-Tg小鼠淋巴結(jié)腫大,其中T細(xì)胞明顯增加,而且活化狀態(tài)增強(qiáng),分泌炎性細(xì)胞因子IFN-γ的能力也增強(qiáng)。進(jìn)一步探究Il2ra-/-Tg小鼠淋巴結(jié)腫大的原因時(shí)我們發(fā)現(xiàn),該小鼠Tfh細(xì)胞的比例和數(shù)量明顯增加,同時(shí)生發(fā)中心B細(xì)胞也明顯增多,淋巴結(jié)中漿細(xì)胞明顯增多,而且非淋巴組織中和出現(xiàn)了漿細(xì)胞的浸潤。我們發(fā)現(xiàn)其外周Treg比例降低而胸腺Treg比例升高。I/2ra-/-Tg小鼠Treg細(xì)胞處于活化狀態(tài),表達(dá)更高水平的Treg功能分子和活化性分子。同時(shí),胸腺Treg體外抑制功能降低而外周Treg抑制功能正常。Il2ra-/-Tg小鼠Treg細(xì)胞高表達(dá)Th1型趨化因子受體CXCR3和轉(zhuǎn)錄因子Eomes,同時(shí)具有更強(qiáng)的IFN-γ分泌能力,TSDR區(qū)域的去甲基化水平也升高。我們發(fā)現(xiàn)Il2ra-/-Tg小鼠胸腺及外周Treg上Nrp-1和PD-1的表達(dá)明顯降低,伴隨著的是Tfr細(xì)胞的消失。利用骨髓嵌合實(shí)驗(yàn)補(bǔ)充正常的Tfr細(xì)胞能抑制Il2ra-/-Tg小鼠Tfh細(xì)胞的增加及生發(fā)中心反應(yīng)的增強(qiáng),說明Tfr的缺陷是小鼠淋巴結(jié)腫大及生發(fā)中心反應(yīng)增強(qiáng)的主要原因。綜上所述,我們發(fā)現(xiàn)了 TGF-β和IL-2信號(hào)共同作用調(diào)節(jié)了 Nrp-1+Treg細(xì)胞以及Tfr細(xì)胞的發(fā)育,為生發(fā)中心依賴的自身抗體介導(dǎo)的自身免疫性疾病的治療提供了新靶點(diǎn)。第二部分自身免疫性骨髓纖維化的發(fā)病機(jī)制研究骨髓纖維化是一種髓系細(xì)胞增生性腫瘤,其特征包括干細(xì)胞來源的髓系細(xì)胞克隆增殖、骨髓成纖維細(xì)胞增多和網(wǎng)狀纖維分布導(dǎo)致的骨髓纖維化、貧血、脾腫大、髓外造血等。通常情況下,骨髓纖維化并不被認(rèn)為是一種自身免疫性疾病。但是在一些情況下,骨髓纖維化可能伴隨自身免疫病出現(xiàn),尤其是系統(tǒng)性紅斑狼瘡(SLE),稱之為自身免疫性骨髓纖維化。自身免疫性骨髓纖維化可伴隨原發(fā)性膽汁性膽管炎(PBC),但其發(fā)生機(jī)制并不清楚。我們前期發(fā)現(xiàn)p40-/-IL-2Rαc-/-模型小鼠是很好的PBC模型小鼠,并會(huì)自發(fā)出現(xiàn)肝臟的纖維化。而我們進(jìn)一步發(fā)現(xiàn),模型小鼠會(huì)出現(xiàn)脾臟腫大、貧血等現(xiàn)象,與臨床骨髓纖維化癥狀相似。因此我們探究了p40-/-IL-2Rα-/-模型小鼠作為自身免疫性骨髓纖維化的模型的可能性,并進(jìn)一步探究及其發(fā)病機(jī)制。不同于對(duì)照小鼠,p40-/-IL-2Rα-/-鼠脾臟出現(xiàn)類似CFU-S實(shí)驗(yàn)的結(jié)節(jié),有大量巨核細(xì)胞,并且肝臟出現(xiàn)血島。在小鼠的外周血、肝臟、脾臟中均出現(xiàn)了造血干細(xì)胞(LSK)樣的細(xì)胞,說明髓外造血的存在。這些升高的LSK細(xì)胞與小鼠脾臟重量及肝臟的炎癥程度成正相關(guān)。與對(duì)照鼠小鼠相比,血液中紅細(xì)胞計(jì)數(shù)、血紅蛋白、血球容積、白細(xì)胞計(jì)數(shù)均明顯降低,說明出現(xiàn)了貧血的情況。HE染色發(fā)現(xiàn)p40-/-IL-2Rα-/-鼠骨髓會(huì)出現(xiàn)成纖維細(xì)胞的增多,銀染發(fā)現(xiàn)骨髓會(huì)有大量網(wǎng)狀纖維,說明小鼠出現(xiàn)了骨髓纖維化。骨髓中LSK樣細(xì)胞增多,且造血能力降低。p40-/-IL-2Rα-/-鼠骨髓浸潤大量活化狀態(tài)的CD4+和CD8+ T細(xì)胞,這些T細(xì)胞有更強(qiáng)的IFN-y分泌能力且與LSK細(xì)胞比例呈正相關(guān)。利用轉(zhuǎn)基因小鼠我們發(fā)現(xiàn),敲除CD4并不會(huì)減輕小鼠的骨髓纖維化,但是敲除CD8a或者敲除IFN-γ都可以阻止骨髓纖維化的發(fā)生,表現(xiàn)為髓外造血消失,骨髓造血干細(xì)胞恢復(fù)正常,骨髓成纖維細(xì)胞消失以及無網(wǎng)狀纖維分布。關(guān)鍵的是,在小鼠發(fā)病之后利用抗CD8α抗體清除CD8+T細(xì)胞,能顯著減輕小鼠自身免疫性骨髓纖維化的發(fā)生。總之,我們發(fā)現(xiàn)CD8+ T細(xì)胞及其產(chǎn)生的細(xì)胞因子IFN-γ是導(dǎo)致小鼠自身免疫性骨髓纖維化的關(guān)鍵因素。利用抗體清除CD8+ T細(xì)胞能對(duì)自身免疫性骨髓纖維化進(jìn)行治療,為臨床上治療提供新的思路。
[Abstract]:The first part studies revealed the cytokines regulate the immune tolerance induced by the factors for the occurrence and development of breaking the understanding of autoimmune diseases have an important role in.Treg cell is negative control cell body immune oerreaction tonality to prevent autoimmune disease of regulatory T cell development, more and more studies show that Treg is also involved in bacteria and virus infection, tumor, organ transplantation tolerance, tissue repair and other diseases. Therefore, is conducive to further understanding of Treg function on Treg development regulation, so it can be used as a therapeutic target in.Treg cells of various diseases can be caused by the development of thymus and peripheral in two ways, the differentiation and development by regulating cell the dominant factor. The inactivation of the TGF- beta type II receptor dnTGF- beta R II in mice caused by excessive activation of T cells in the liver and the expression of CD4 promoter Intestinal inflammatory cell infiltration, inhibit the function of Treg cells in.Il2ra-/- mice reduced disorders due to the Treg function, the amplification of polyclonal T cells and B cells and cause multiple organ autoimmune diseases. In this study, we used Il2ra-/-Tg mice, regulated by exploring the role of TGF- beta and IL-2 signal in Treg development process. We found that Il2ra-/-Tg mice showed symptoms similar to scurfy mice deficient in Treg, multiple organs appeared lymphocytic infiltration and pathological enlargement of lymph nodes of mice.Il2ra-/-Tg, T cells were significantly increased, and the activation state of enhanced secretion of inflammatory cytokines IFN- gamma is also enhanced. To further explore the causes of lymphadenopathy in Il2ra-/-Tg mice we found that the proportion of Tfh cells in mice and significantly increased the number at the same time, the germinal center B cells also increased significantly, the lymph node of plasma cells increased significantly, and And non lymphoid tissue and the infiltration of plasma cells. We found that Treg decreased the proportion of peripheral and thymus Treg increased.I/2ra-/-Tg activation in mouse Treg cells, the expression of Treg molecules and higher levels of activation of molecules. At the same time, the thymus Treg in vitro inhibition function decreased peripheral Treg inhibited expression of Th1 chemokines CXCR3 receptor and transcription factor Eomes of Treg cells in mice with normal.Il2ra-/-Tg function, IFN- also has a stronger gamma secretion capacity, TSDR region demethylation level also increased. We found that the expression of Nrp-1 and PD-1 and Il2ra-/-Tg week Treg mice thymus was significantly decreased, with the disappearance of Tfr cells. The bone marrow chimeric experiments normal Tfr cells can inhibit the Il2ra-/-Tg of mouse Tfh cells increased and the germinal center reaction enhancement, Tfr defects is the lymph nodes and in mice The main reason was enhanced. In summary, we found that the interaction of TGF- beta and IL-2 signal regulated Nrp-1+Treg cell and Tfr cell development, provides a new therapeutic target for autoantibody induced germinal center dependent mediated autoimmune diseases. The second part of the pathogenesis of autoimmune bone marrow fibrosis fibrosis is a myeloid proliferative tumor, characterized by clonal proliferation of myeloid cells from stem cells, bone marrow fibroblasts increased and reticular fiber distribution leads to bone marrow fibrosis, anemia, splenomegaly, extramedullary hematopoiesis. Typically, bone marrow fibrosis is not considered an autoimmune disease but in some cases, bone marrow fibrosis may be associated with autoimmune diseases, especially systemic lupus erythematosus (SLE), called autoimmune bone marrow fibrosis itself. Autoimmune myelofibrosis with primary biliary cholangitis (PBC), but its pathogenesis is not clear. We previously found that the p40-/-IL-2R alpha c-/- mouse model is a good model of PBC mice, and spontaneously liver fibrosis. And we further found that the mouse model will appear splenomegaly, anemia and other phenomena. Similar to the clinical symptoms of bone marrow fibrosis. So we study the p40-/-IL-2R - / - mice alpha possibility as autoimmune bone marrow fibrosis model, and further explore its pathogenesis. Different from control mice, spleen p40-/-IL-2R alpha - rat CFU-S like experimental nodules, there are a large number of megakaryocytes, and liver blood liver island. In the peripheral blood of mice, spleen occurred in hematopoietic stem cells (LSK) like cells, indicating extramedullary hematopoiesis. These elevated LSK cells and spleen weight in mice The degree of inflammation and liver are positively correlated. Compared with control mice, red blood cell count, blood hemoglobin, hematocrit and leukocyte count were significantly decreased, indicating the anemia.HE staining indicated that p40-/-IL-2R alpha - there will be increased into bone marrow fibroblasts, silver staining showed that bone marrow may have a large network fiber, that mice showed bone marrow fibrosis. LSK like cells in the bone marrow of.P40-/-IL-2R rats was increased, a large bone marrow infiltration of the activation state of CD4+ and CD8+ in T cells decreased and hematopoietic ability, these T cells have stronger IFN-y secretion ability and positively related with the percentage of LSK cells in transgenic mice. We found that knockdown of CD4 and do not reduce the mice bone marrow fibrosis, but CD8a knockdown or knockout of IFN- gamma can prevent bone marrow fibrosis, manifested as extramedullary hematopoiesis disappeared, bone marrow hematopoietic stem cells to restore normal bone. Pulp fibroblast cells disappeared and no reticular fiber distribution. The key is that the use of anti CD8 antibody depletion of CD8+T cells in mice after onset, can significantly reduce the mice autoimmune bone marrow fibrosis. In short, we found that the cytokines IFN- CD8+ T cells and the key factors leading to autoimmune myelofibrosis the use of antibodies to remove CD8+ T. Cells can treat autoimmune myelofibrosis, to provide new ideas for clinical treatment.

【學(xué)位授予單位】：中國科學(xué)技術(shù)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R593.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 鄒小立 ,黃梓倫;骨髓纖維化[J];新醫(yī)學(xué);2001年11期

2 王玉林,李剛;骨髓纖維化合并食管靜脈曲張破裂出血1例[J];廣東醫(yī)學(xué);2001年07期

3 周家善,任紅波,吳斌,周正菊,袁承泰;高嗜酸粒細(xì)胞綜合征并骨髓纖維化一例[J];中華醫(yī)學(xué)雜志;2002年06期

4 居乃新,丁明巖;以腹水為首發(fā)癥狀的骨髓纖維化1例[J];中國實(shí)用內(nèi)科雜志;2003年04期

5 賴洵;聞艷;虞利群;;原發(fā)性骨髓纖維化合并自體免疫性溶血性貧血1例分析[J];中國誤診學(xué)雜志;2006年09期

6 孫志新;;急性粒細(xì)胞白血病并骨髓纖維化一例報(bào)告[J];青海醫(yī)藥;1984年04期

7 陳輝樹;甘芾;梁曉嵐;儲(chǔ)榆林;;骨髓纖維化臨床與病理分析—關(guān)于分型問題的討論[J];天津醫(yī)藥;1985年12期

8 郭瑞官;林莘野;李健;陳銑;黃明清;;小兒骨髓纖維化(附2例報(bào)告)[J];福建醫(yī)學(xué)院學(xué)報(bào);1988年02期

9 高翔;鄭元龍;;骨髓纖維化二例[J];泰山醫(yī)學(xué)院學(xué)報(bào);1988年04期

10 孫桂珍;周建國;;原發(fā)性血小板增多癥轉(zhuǎn)化骨髓纖維化及急性粒一單核細(xì)胞白血病1例報(bào)告[J];上海醫(yī)學(xué);1992年07期

相關(guān)會(huì)議論文 前7條

1 楊波;高芳;李建蘭;魏艷紅;喬巨;;骨髓增生異常綜合征合并骨髓纖維化的臨床及形態(tài)學(xué)分析[A];第一屆全國難治性淋巴瘤學(xué)術(shù)研討會(huì)暨第二屆全國多發(fā)性骨髓瘤學(xué)術(shù)研討會(huì)（國家級(jí)血液系統(tǒng)惡性腫瘤繼續(xù)醫(yī)學(xué)教育學(xué)習(xí)班）繼續(xù)教育集[C];2007年

2 宋國緒;;血清陰性脊柱關(guān)節(jié)病合并骨髓纖維化1例報(bào)告[A];海峽兩岸中醫(yī)藥發(fā)展大會(huì)風(fēng)濕論文集[C];2009年

3 錢美華;;骨髓纖維化診治進(jìn)展[A];2009年浙江省中醫(yī)藥學(xué)會(huì)血液病學(xué)術(shù)年會(huì)、浙江省中西醫(yī)結(jié)合學(xué)會(huì)血液病學(xué)術(shù)年會(huì)暨國家級(jí)中西醫(yī)結(jié)合血液病新進(jìn)展繼續(xù)教育學(xué)習(xí)班論文匯編[C];2009年

4 楊波;高芳;李建蘭;楊林花;喬振華;;骨髓增生異常綜合征伴骨髓纖維化的形態(tài)學(xué)與臨床分析[A];中華醫(yī)學(xué)會(huì)第七次全國檢驗(yàn)醫(yī)學(xué)學(xué)術(shù)會(huì)議資料匯編[C];2008年

5 陳幸華;張曦;張誠;高蕾;孔佩艷;彭賢貴;劉紅;王慶余;;自體外周血干細(xì)胞移植治療急性單核細(xì)胞白血病并骨髓纖維化1例報(bào)道[A];第11次中國實(shí)驗(yàn)血液學(xué)會(huì)議論文匯編[C];2007年

6 劉鳳珍;孫熠鋒;靳洪濤;郭惠芳;邵福靈;;結(jié)締組織病相關(guān)性骨髓纖維化[A];第十屆全國風(fēng)濕病學(xué)學(xué)術(shù)會(huì)議論文集[C];2005年

7 羊紅玉;;骨髓纖維化新藥Ruxolitinib藥理及臨床評(píng)價(jià)[A];2011年浙江省醫(yī)學(xué)會(huì)臨床藥學(xué)分會(huì)學(xué)術(shù)年會(huì)論文匯編[C];2011年

相關(guān)重要報(bào)紙文章 前4條

1 ;中藥可抑制骨髓纖維化[N];河北科技報(bào)(農(nóng)村版);2000年

2 吳維海;從肝論治骨髓纖維化[N];健康報(bào);2004年

3 石家莊平安醫(yī)院主任醫(yī)師 吳維海;我們用中醫(yī)治療骨髓纖維化[N];健康報(bào);2005年

4 辛藍(lán);諾華與Incyte公司達(dá)成藥物授權(quán)協(xié)議[N];醫(yī)藥經(jīng)濟(jì)報(bào);2009年

相關(guān)博士學(xué)位論文 前2條

1 李亮;細(xì)胞因子對(duì)調(diào)節(jié)性T細(xì)胞發(fā)育調(diào)節(jié)及自身免疫性骨髓纖維化發(fā)病機(jī)制研究[D];中國科學(xué)技術(shù)大學(xué);2017年

2 徐俊卿;1、原發(fā)性骨髓纖維化患者脾腫大的預(yù)后意義 2、真性紅細(xì)胞增多癥和原發(fā)性血小板增多癥后骨髓纖維化患者的臨床特征研究 3、小劑量沙利度胺和潑尼松聯(lián)合或不聯(lián)合達(dá)那唑治療原發(fā)性骨髓纖維化患者貧血的療效比較 4、骨髓增殖性腫瘤患者生活質(zhì)量評(píng)估[D];北京協(xié)和醫(yī)學(xué)院;2014年

相關(guān)碩士學(xué)位論文 前3條

1 潘成云;乏氧環(huán)境下LOXL-2刺激MSC活化為腫瘤相關(guān)成纖維細(xì)胞促進(jìn)MPN骨髓纖維化進(jìn)程研究[D];南方醫(yī)科大學(xué);2017年

2 邢麗娜;原發(fā)性血小板增多癥發(fā)病機(jī)理與診斷治療研究進(jìn)展[D];河北醫(yī)科大學(xué);2009年

3 楊海英;一例骨髓纖維化轉(zhuǎn)化為急性髓細(xì)胞白血病的診斷思路[D];山西醫(yī)科大學(xué);2007年

，

本文編號(hào)：1561518