本文選題：系統(tǒng)性紅斑狼瘡 + B細(xì)胞��； 參考：《南京大學(xué)》2016年碩士論文

【摘要】：系統(tǒng)性紅斑狼瘡(Systemic lupus erythematosus, SLE)是一種慢性的、累及多器官多系統(tǒng)的自身免疫性疾病,其主要特征是患者血清中存在大量的IFN-α和自身抗體。B細(xì)胞作為抗體產(chǎn)生細(xì)胞,其異常活化在SLE的發(fā)病過程中發(fā)揮關(guān)鍵作用,然而其異�；罨姆肿訖C(jī)制仍不完全清楚。許多研究報(bào)道,SLE的B細(xì)胞中IFN-α通路是高度活化的,且IFN-α可通過直接或間接的方式參與B細(xì)胞的生存與功能,進(jìn)而參與SLE的病程發(fā)展；在SLE患者體內(nèi),CD180-B細(xì)胞數(shù)量明顯增加,并且參與自身抗體的產(chǎn)生,提示CD180可能參與SLE中B細(xì)胞的異�；罨�。那么,CD180的表達(dá)變化是否影響IFN-α通路活化?導(dǎo)致CD180表達(dá)變化的原因又是什么?因此,本研究針對(duì)上述問題,分析了CD180在SLE來源B細(xì)胞的表達(dá)變化,鑒定了CD180-B細(xì)胞的表型,探討了CD180調(diào)控IFN-α通路影響B(tài)細(xì)胞的分子機(jī)制。1.發(fā)現(xiàn)SLE來源B細(xì)胞低表達(dá)CD180,且CD180-B細(xì)胞可能由生發(fā)中心B細(xì)胞和漿細(xì)胞組成；我們首先各取5例正常人與SLE患者外周血進(jìn)行流式分析,驗(yàn)證SLE患者外周血CD180-B細(xì)胞異常增多；從分選出B細(xì)胞后檢測(cè)CD180 mRNA水平,發(fā)現(xiàn)CD180表達(dá)水平在SLE患者中是顯著降低的。表明CD180-B細(xì)胞的增多可能是由于某些原因?qū)е碌腂細(xì)胞CD180表達(dá)降低有關(guān)。接下來我們?cè)诶钳從Ｐ托∈笾羞M(jìn)行了驗(yàn)證,發(fā)現(xiàn)在狼瘡小鼠脾臟、外周血和骨髓中,CD180-B細(xì)胞數(shù)量都是明顯增加,且分選出的脾臟B細(xì)胞的CD180表達(dá)降低。我們進(jìn)一步研究了CD180-B細(xì)胞的表型,發(fā)現(xiàn)CD180-B細(xì)胞可能由生發(fā)中心B細(xì)胞和漿細(xì)胞組成。2.發(fā)現(xiàn)激活CD180能夠顯著抑制B細(xì)胞的IFN-α通路活化,提示anti-CD180可能是治療SLE的潛在策略：我們探討了CD180與IFN-α通路的關(guān)系及影響其表達(dá)變化的原因,發(fā)現(xiàn)激活CD180在體外和體內(nèi)均能顯著抑制B細(xì)胞IFN-α通路的活化并降低IFN-α下游基因的表達(dá)。其具體機(jī)制是CD180通過lyn-PI3K-BTK這一級(jí)聯(lián)通路引起IFN-α下游STAT-2磷酸化水平下降,進(jìn)而影響IFN-α通路的活化。進(jìn)一步發(fā)現(xiàn),用TLR7配體R848和TLR9配體CpG刺激小鼠脾臟B細(xì)胞能顯著抑制CD180的表達(dá),并調(diào)節(jié)CD180對(duì)IFN-α通路的抑制作用。綜上所述,我們證實(shí)了MRL/lpr小鼠中CD180-B細(xì)胞顯著增多；發(fā)現(xiàn)激活CD180能夠顯著抑制IFN-α通路的活化；還發(fā)現(xiàn)SLE來源B細(xì)胞CD180的表達(dá)變化可能歸因于TLR7和TLR9通路活化。由于靶向IFN-α治療SLE具有很大的前景,我們的發(fā)現(xiàn)提示,anti-CD 180可能是治療SLE的潛在策略。
[Abstract]:Systemic lupus erythematosus (lupus erythematosus, SLE) is a chronic, multi-organ and multi-system autoimmune disease characterized by the presence of a large number of IFN- 偽 and autoantibody. B cells as antibody producing cells. Its abnormal activation plays a key role in the pathogenesis of SLE, but the molecular mechanism of its abnormal activation is still unclear. Many studies have reported that IFN- 偽 pathway is highly activated in B cells of SLE, and IFN- 偽 can participate in the survival and function of B cells directly or indirectly, and then participate in the course of SLE, and the number of CD180-B cells in patients with SLE increases obviously. It is suggested that CD180 may be involved in the abnormal activation of B cells in SLE. Does the expression of CD180 affect the activation of IFN- 偽 pathway? What is the cause of the change in CD180 expression? Therefore, in this study, we analyzed the expression of CD180 in SLE derived B cells, identified the phenotype of CD180-B cells, and explored the molecular mechanism of CD180 regulating IFN- 偽 pathway affecting B cells. It was found that SLE derived B cells were low expressed CD180, and CD180-B cells might be composed of germinal center B cells and plasma cells. Firstly, we took 5 normal persons and 5 SLE patients for flow analysis to verify the abnormal increase of CD180-B cells in peripheral blood of SLE patients. The level of CD180 mRNA was detected from B cells, and the expression of CD180 was significantly decreased in SLE patients. The results suggest that the increase of CD180-B cells may be related to the decrease of CD180 expression in B cells. Then we demonstrated that the number of CD180-B cells in spleen, peripheral blood and bone marrow of lupus mice was significantly increased, and the CD180 expression of selected B cells in spleen was decreased. We further studied the phenotype of CD180-B cells and found that CD180-B cells may consist of germinal center B cells and plasma cells. It was found that activation of CD180 could significantly inhibit the activation of IFN- 偽 pathway in B cells, suggesting that anti-CD180 might be a potential strategy for the treatment of SLE. We investigated the relationship between CD180 and IFN- 偽 pathway and the causes of its expression changes. It was found that activation of CD180 could significantly inhibit the activation of IFN- 偽 pathway and decrease the expression of IFN- 偽 downstream gene in B cells in vitro and in vivo. The mechanism is that CD180 leads to the decrease of phosphorylation level of STAT-2 downstream of IFN- 偽 through lyn-PI3K-BTK, which affects the activation of IFN- 偽 pathway. It was further found that TLR7 ligand R848 and TLR9 ligand CpG could significantly inhibit the expression of CD180 and regulate the inhibitory effect of CD180 on IFN- 偽 pathway. In conclusion, we confirmed that the number of CD180-B cells in MRL/lpr mice was significantly increased, the activation of CD180 could significantly inhibit the activation of IFN- 偽 pathway, and the change of CD180 expression in SLE derived B cells might be attributed to the activation of TLR7 and TLR9 pathway. Our findings suggest that anti-CD180 may be a potential strategy for the treatment of SLE.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R593.241

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