本文選題：immature + B細(xì)胞��； 參考：《第四軍醫(yī)大學(xué)》2008年博士論文

【摘要】： 目的:B淋巴細(xì)胞起源于多能造血干細(xì)胞,在整個(gè)發(fā)育過程當(dāng)中經(jīng)歷了多個(gè)發(fā)育階段,造血干細(xì)胞必須成功經(jīng)過各個(gè)發(fā)育階段后,才能成為一個(gè)正常發(fā)揮免疫應(yīng)答功能的成熟B淋巴細(xì)胞。但是B細(xì)胞發(fā)育的精確調(diào)控機(jī)制可能會(huì)因許多病理?xiàng)l件而被擾亂,特別值得關(guān)注的是炎癥的影響。Toll樣受體是一種能夠識(shí)別多種病原相關(guān)分子的蛋白,一旦識(shí)別病原相關(guān)分子之后,它們可以啟動(dòng)一系列反應(yīng)激活免疫系統(tǒng)清除入侵病原微生物,從而構(gòu)成機(jī)體免疫的第一道防線;TLR誘導(dǎo)產(chǎn)生炎性細(xì)胞因子,而許多這類因子對B細(xì)胞發(fā)育均有抑制作用。這兩個(gè)原本獨(dú)立的現(xiàn)象提示TLR信號(hào)與B細(xì)胞發(fā)育間存在某種關(guān)聯(lián)。考慮到TLR信號(hào)對成熟B細(xì)胞功能具有直接而深刻的影響,一個(gè)顯而易見的問題是TLR信號(hào)是否對發(fā)育中的B細(xì)胞也有類似的直接作用。在本試驗(yàn)中,我們檢測了TLR4在各個(gè)階段B細(xì)胞中的表達(dá)水平,檢測了正常C3H/He小鼠和C3H/HeJ小鼠之間B細(xì)胞分布的區(qū)別,同時(shí)通過體內(nèi)和體外試驗(yàn)檢測了TLR4信號(hào)對發(fā)育中的B細(xì)胞的影響。 方法:通過實(shí)時(shí)熒光定量PCR檢測TLR4在各個(gè)B細(xì)胞亞群中的表達(dá)水平;對C57BL/6小鼠腹膜內(nèi)注射LPS,在不同時(shí)間點(diǎn)取骨髓通過細(xì)胞計(jì)數(shù)及流式檢測B細(xì)胞及各個(gè)亞群的變化;流式細(xì)胞儀分析正常C3H/He小鼠和C3H/HeJ小鼠骨髓內(nèi)全B細(xì)胞以及B細(xì)胞亞群的比例; FACs分選出的未成熟B細(xì)胞體外培養(yǎng),LPS處理后通過CFSE染色檢測增殖代數(shù),通過流式檢測培養(yǎng)細(xì)胞表面IgD、CD21、CD23、CD86的表達(dá);FACs分選出的未成熟B細(xì)胞體外培養(yǎng),分別用LPS、anti-μ或者LPS+anti-μ處理之后,流式檢測細(xì)胞分化和凋亡的相關(guān)指標(biāo)。 結(jié)果:(1)實(shí)時(shí)熒光定量PCR結(jié)果顯示小鼠骨髓中四個(gè)B細(xì)胞亞群均表達(dá)TLR4,并且mature B細(xì)胞的表達(dá)水平明顯高于前三個(gè)亞群,而脾來源的mature B細(xì)胞的表達(dá)水平又明顯高于骨髓來源的mature B細(xì)胞;(2)對C57BL/6小鼠應(yīng)用LPS處理后,發(fā)現(xiàn)處理后的小鼠骨髓細(xì)胞總數(shù)、全B細(xì)胞、pre B細(xì)胞和immature B細(xì)胞計(jì)數(shù)在第1周會(huì)明顯下降,隨后會(huì)出現(xiàn)反彈性的異常增高,在第3周的時(shí)候逐漸恢復(fù)正常水平;(3)在C3H/HeJ小鼠骨髓中,全B細(xì)胞、pro B、pre B細(xì)胞占總骨髓細(xì)胞的比例均明顯高于正常的C3H/He小鼠,而immature B和mature B細(xì)胞的比例沒有統(tǒng)計(jì)學(xué)意義的差別;(4)Immature B細(xì)胞體外培養(yǎng)體系中應(yīng)用LPS刺激后,IgD、CD21、CD23、CD86的陽性率均明顯高于陰性對照組;(5)Immature B細(xì)胞體外培養(yǎng)體系中應(yīng)用LPS、anti-μ或者LPS+anti-μ刺激后,發(fā)現(xiàn)LPS+anti-μ組的凋亡細(xì)胞比例明顯低于anti-μ組,而其IgD陽性率則明顯低于LPS組。 結(jié)論:(1)TLR4的表達(dá)水平在新生B淋巴細(xì)胞遷出骨髓后會(huì)有一個(gè)大幅度的提高,這種提高可能是受到外周環(huán)境當(dāng)中的微生物組分和各種細(xì)胞因子的影響;(2)當(dāng)小鼠體內(nèi)TLR4信號(hào)過強(qiáng)時(shí),骨髓內(nèi)的B淋巴細(xì)胞也會(huì)出現(xiàn)異常發(fā)育,至于這種異常是由于早期造血干細(xì)胞向淋系的發(fā)育受阻還是由于B淋巴細(xì)胞的發(fā)育加速,經(jīng)過我們的分析,這兩種原因可能在這一過程中都起著一定的作用;(3)在TLR4功能缺失的小鼠體內(nèi)B淋巴細(xì)胞的發(fā)育是異常的,主要是阻滯在pro B、pre B和immature B細(xì)胞三個(gè)早期的階段;(4)TLR4信號(hào)可以使immature B細(xì)胞向成熟方向分化,并可以幫助自身免疫性B淋巴細(xì)胞逃避陰性選擇過程免于凋亡;(5)LPS促進(jìn)immature B細(xì)胞分化成熟這一作用可以被anti-μ抑制掉。以上試驗(yàn)結(jié)果提示TLR4信號(hào)在體內(nèi)外均可促進(jìn)immature B細(xì)胞的發(fā)育,而機(jī)體產(chǎn)生B淋巴細(xì)胞是一個(gè)極其復(fù)雜的過程,受著多種不同因素的影響。
[Abstract]:Objective: B lymphocytes originate from multipotent hematopoietic stem cells and undergo multiple developmental stages during the whole development process. The hematopoietic stem cells must successfully pass through various developmental stages to become a mature B lymphocyte that normally plays the immune response function, but the precise regulation mechanism of B cell development may be caused by many pathological strips. The.Toll like receptor is a protein that recognizes a variety of pathogenic molecules. Once the pathogenic molecules are identified, they can activate a series of responses to activate the immune system to clear the pathogenic microbes, thus forming the first line of defense for the immune system; TLR induced production. Inflammatory cytokines, and many of these factors inhibit the development of B cells. These two original independent phenomena suggest that there is a certain association between the TLR signal and the development of B cells. Considering that the TLR signal has a direct and profound effect on the function of mature B cells, the obvious question is whether the TLR signal is also the B cells in the developing B cells. In this experiment, we detected the expression level of TLR4 in B cells at various stages, detected the difference in the distribution of B cells between normal C3H/He mice and C3H/HeJ mice, and detected the effect of TLR4 signals on the developing B cells by both in vivo and in vitro tests.
Methods: the expression level of TLR4 in each B cell subgroup was detected by real time fluorescence quantitative PCR; LPS was injected into the peritoneum of C57BL/6 mice. The changes of the bone marrow through cell count and flow cytometry were taken at different time points. The flow cytometry was used to analyze the total B and B cells in the bone marrow of normal C3H/He mice and C3H/HeJ mice. The proportion of subgroups; the immature B cells selected by FACs were cultured in vitro, and the proliferation algebra was detected by CFSE staining after LPS treatment. The expression of IgD, CD21, CD23 and CD86 on the surface of the cells was cultured by flow cytometry, and the immature B cells isolated by FACs were cultured in vitro, and the cell differentiation and withering were detected with LPS, anti-, or micron, respectively. Related indicators of death.
Results: (1) real time fluorescence quantitative PCR results showed that four B cell subsets in mouse bone marrow expressed TLR4, and the expression level of mature B cells was significantly higher than that of the first three subgroups, while the expression level of mature B cells in the spleen was significantly higher than that of mature B cells derived from bone marrow; (2) after the application of LPS treatment to C57BL/6 mice, the treatment was found after treatment. The total number of bone marrow cells, total B cells, pre B cells and immature B cells decreased significantly at first weeks, followed by an abnormal increase in anti elasticity, and gradually restored to normal levels at third weeks. (3) the proportion of all B cells in the bone marrow of C3H/HeJ mice, pro B, pre B cells were significantly higher than that of normal C3H/He. There was no significant difference in the proportion of immature B and mature B cells; (4) the positive rates of IgD, CD21, CD23, CD86 were significantly higher than those in the negative control group after the application of LPS stimulation in the culture system of Immature B cells. The percentage of apoptotic cells was significantly lower than that of group anti-, while the positive rate of IgD was significantly lower than that of LPS group.
Conclusion: (1) the expression level of TLR4 can be greatly improved after the migration of B lymphocytes from the newborn cells, which may be influenced by the microbiological components and various cytokines in the peripheral environment. (2) the abnormal development of the B lymphocytes in the bone marrow is also developed when the TLR4 signal is too strong in the mice, as for this abnormality. It is due to the obstruction of the development of the early hematopoietic stem cells to the drenching system or the acceleration of the development of B lymphocytes. Through our analysis, these two reasons may play a role in this process. (3) the development of B lymphocytes in the mice with TLR4 dysfunction is abnormal, mainly blocking the Pro B, pre B and immature B fine. Three early stages of the cell; (4) the TLR4 signal can differentiate the immature B cells from the mature direction and help the autoimmune B lymphocytes escape the negative selection process from apoptosis; (5) the effect of LPS to promote the differentiation and maturation of immature B cells can be suppressed by anti- us. The results suggested that the TLR4 signal can be promoted in vivo and in vitro. The development of immature B cells and the generation of B lymphocytes in the body are very complex processes, which are affected by many different factors.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳國慶;;運(yùn)動(dòng)對骨骼肌衛(wèi)星細(xì)胞的影響及作用機(jī)制[J];四川解剖學(xué)雜志;2011年02期

2 盧明;;神經(jīng)膠質(zhì)細(xì)胞對神經(jīng)干細(xì)胞增殖和分化影響的研究進(jìn)展[J];湖南師范大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2011年02期

3 翟遠(yuǎn)坤;李志忠;陳克明;張娜;程國政;朱瑞清;;淫羊藿苷對體外培養(yǎng)乳鼠顱骨成骨細(xì)胞增殖、分化及成熟的影響[J];中藥材;2011年06期

4 杞少華;武棟成;李東升;;MicroRNA在干細(xì)胞增殖與分化過程中的調(diào)控作用[J];中國組織工程研究與臨床康復(fù);2011年27期

5 劉偉;趙勁民;蘇偉;李曉峰;秦義武;;骨碎補(bǔ)總黃酮可促進(jìn)兔骨髓間充質(zhì)干細(xì)胞的增殖和分化[J];中國組織工程研究與臨床康復(fù);2011年32期

6 謝興文;宋敏;金成強(qiáng);;接骨散含藥血清對骨髓間充質(zhì)干細(xì)胞增殖與成骨性分化影響[J];中國中醫(yī)骨傷科雜志;2011年06期

7 張小潤;張程;王曉宇;張湘燕;李敏;葉賢偉;夏婧;;L-氨基酸氧化酶對人肺腺癌A549細(xì)胞增殖、凋亡的影響研究[J];中國藥房;2011年25期

8 郭玲;鄭立舸;王敏;郝亮;;小鼠成骨細(xì)胞Wnt信號(hào)通路相關(guān)因子表達(dá)與胰島素樣生長因子1的影響[J];中國組織工程研究與臨床康復(fù);2011年28期

9 劉鈺瑜;姚衛(wèi)民;徐道華;崔燎;;卡托普利對MC3T3-E1細(xì)胞增殖、分化及Ⅰ型膠原mRNA表達(dá)的影響[J];中國藥理學(xué)通報(bào);2011年09期

10 陳良建;汪瑞芳;郭小平;李益民;何浩;李挺;;載TGF-β_1明膠微球涂層多孔鈦對MG63細(xì)胞功能的影響[J];中南大學(xué)學(xué)報(bào)(自然科學(xué)版);2011年08期

相關(guān)會(huì)議論文 前10條

1 康樂;姚東璧;胡庭俊;曾蕓;韋英益;李躍華;;天門冬多糖對小鼠脾臟淋巴細(xì)胞體外增殖的影響[A];紀(jì)念中國畜牧獸醫(yī)學(xué)會(huì)中獸醫(yī)學(xué)分會(huì)成立30周年中國畜牧獸醫(yī)學(xué)會(huì)中獸醫(yī)學(xué)分會(huì)2009年學(xué)術(shù)年會(huì)、華東區(qū)第十九次中獸醫(yī)科研協(xié)作與學(xué)術(shù)研討會(huì)論文集[C];2009年

2 朱玲玲;范明;;低氧在中樞神經(jīng)系統(tǒng)發(fā)育中的作用[A];提高全民科學(xué)素質(zhì)、建設(shè)創(chuàng)新型國家——2006中國科協(xié)年會(huì)論文集[C];2006年

3 陳玉;張勇剛;吳秦潔;胡火珍;;小鼠神經(jīng)干細(xì)胞與骨髓間充質(zhì)干細(xì)胞的共培養(yǎng)[A];中國細(xì)胞生物學(xué)學(xué)會(huì)第九次會(huì)員代表大會(huì)暨青年學(xué)術(shù)大會(huì)論文摘要集[C];2007年

4 高瑞蘭;陳小紅;林筱潔;錢煦岱;周郁鴻;徐衛(wèi)紅;;三七皂苷在造血細(xì)胞內(nèi)MAPK信號(hào)傳遞途徑的研究[A];第11次中國實(shí)驗(yàn)血液學(xué)會(huì)議論文匯編[C];2007年

5 姜俊;周小秋;;Gln對鯉魚腸上皮細(xì)胞增殖和分化的影響[A];動(dòng)物營養(yǎng)與飼料研究——第五屆全國飼料營養(yǎng)學(xué)術(shù)研討會(huì)論文集[C];2006年

6 高瑞蘭;林筱潔;陳小紅;徐衛(wèi)紅;錢煦岱;吳超群;;三七皂苷誘導(dǎo)造血細(xì)胞基因表達(dá)譜的實(shí)驗(yàn)研究[A];第11次中國實(shí)驗(yàn)血液學(xué)會(huì)議論文匯編[C];2007年

7 祝青;刁慶春;;聚焦超聲對慢性濕疹模型豚鼠皮膚淋巴細(xì)胞和角質(zhì)形成細(xì)胞凋亡的影響[A];中華醫(yī)學(xué)會(huì)第14次全國皮膚性病學(xué)術(shù)年會(huì)論文匯編[C];2008年

8 高福祿;牛嗣云;龐小靜;陳龍;李秀華;;胎兒睪丸發(fā)生過程中生殖細(xì)胞的增殖與凋亡[A];解剖學(xué)雜志——中國解剖學(xué)會(huì)2002年年會(huì)文摘匯編[C];2002年

9 李奎;李明;康相濤;劉英;;固始雞脾臟內(nèi)淋巴細(xì)胞自然凋亡超微結(jié)構(gòu)研究[A];中國畜牧獸醫(yī)學(xué)會(huì)動(dòng)物解剖學(xué)及組織胚胎學(xué)分會(huì)第十五次學(xué)術(shù)研討會(huì)論文集[C];2008年

10 任思沖;陳鳳秀;尉艷霞;潘虹;何宣霖;王若菡;劉戟;;洋蔥中的黃酮類物質(zhì)對人結(jié)腸癌HCT116細(xì)胞增殖及凋亡的影響[A];第九屆西南三省一市生物化學(xué)與分子生物學(xué)學(xué)術(shù)交流會(huì)論文集[C];2008年

相關(guān)重要報(bào)紙文章 前10條

1 記者　馬霞 實(shí)習(xí)生 張學(xué)琦;專家研討共軛亞油酸免疫功效[N];科技日報(bào);2006年

2 靳共元;關(guān)于“增值”與“增殖”[N];山西經(jīng)濟(jì)日報(bào);2002年

3 盧蘇燕;T淋巴細(xì)胞“作戰(zhàn)”路線被成功拍攝[N];醫(yī)藥經(jīng)濟(jì)報(bào);2007年

4 西南證券 毛祖宏;反彈后的分化[N];中國有色金屬報(bào);2002年

5 ;曾凡勝 窄幅震蕩 板塊分化[N];中國信息報(bào);2000年

6 李紅;互聯(lián)網(wǎng)面臨第二次分化？[N];中國企業(yè)報(bào);2001年

7 國信證券趙秋燕;震蕩加劇　分化明顯[N];中華工商時(shí)報(bào);2002年

8 陸文;凋亡細(xì)胞 被擠出上皮組織[N];醫(yī)藥經(jīng)濟(jì)報(bào);2002年

9 韓朝華;“增值”與“增殖”[N];中國社會(huì)科學(xué)院院報(bào);2003年

10 上證聯(lián);熱點(diǎn)分化 震蕩加劇[N];山西經(jīng)濟(jì)日報(bào);2001年

相關(guān)博士學(xué)位論文 前10條

1 韓冬梅;TLR4信號(hào)調(diào)節(jié)小鼠B淋巴細(xì)胞發(fā)育的實(shí)驗(yàn)研究[D];第四軍醫(yī)大學(xué);2008年

2 仇麗鴻;靜磁場對大鼠成骨細(xì)胞及其牙周炎組織影響的研究[D];中國醫(yī)科大學(xué);2004年

3 羅曉光;小膠質(zhì)細(xì)胞對神經(jīng)損傷環(huán)境中MSCs生物學(xué)行為的影響及其信號(hào)傳導(dǎo)通路初探[D];中國醫(yī)科大學(xué);2006年

4 王文清;棉酚對血液腫瘤細(xì)胞系的殺傷分化效應(yīng)及機(jī)制研究[D];第四軍醫(yī)大學(xué);2006年

5 田海濱;小鼠和山羊胚胎干細(xì)胞建系及其分化為神經(jīng)元細(xì)胞的基礎(chǔ)研究[D];山東大學(xué);2005年

6 張文瑾;早期凋亡細(xì)胞炎癥及免疫抑制性質(zhì)的實(shí)驗(yàn)研究[D];浙江大學(xué);2005年

7 鎮(zhèn)海文;LPS調(diào)節(jié)TLR4介導(dǎo)促進(jìn)心肌細(xì)胞存活時(shí)間的實(shí)驗(yàn)研究[D];南京醫(yī)科大學(xué);2011年

8 王居平;TLR4乙酰化/甲基化修飾激活對炎癥免疫的調(diào)節(jié)[D];浙江大學(xué);2011年

9 吳雄志;骨髓間充質(zhì)干細(xì)胞在肝損傷模型小鼠體內(nèi)向肝細(xì)胞分化[D];四川大學(xué);2005年

10 張學(xué)進(jìn);眼眶前脂肪細(xì)胞在甲狀腺相關(guān)眼病發(fā)病機(jī)理中作用的研究[D];四川大學(xué);2004年

相關(guān)碩士學(xué)位論文 前10條

1 陳步遠(yuǎn);黃芩苷對HL-60細(xì)胞增殖、凋亡、分化的影響及其作用機(jī)制的探討[D];福建醫(yī)科大學(xué);2006年

2 劉蓓;活化TLR4誘導(dǎo)單核細(xì)胞性白血病THP-1細(xì)胞凋亡機(jī)制的研究[D];中南大學(xué);2007年

3 劉陽;ConA的胰蛋白酶消化產(chǎn)物對培養(yǎng)家兔靜息軟骨細(xì)胞的增殖分化影響[D];吉林大學(xué);2006年

4 李麗;褪黑素對人骨髓間充質(zhì)干細(xì)胞的作用研究及其機(jī)制初步探討[D];山西醫(yī)科大學(xué);2007年

5 羅婷婷;維胺酯對角質(zhì)形成細(xì)胞增殖和分化的影響及光致產(chǎn)物的LC-MS分析[D];中南大學(xué);2007年

6 林紅;黃芩苷對CEM白血病細(xì)胞株增殖、凋亡的影響及其機(jī)制的初步研究[D];福建醫(yī)科大學(xué);2008年

7 陳粉粉;EGCG對豬前體脂肪細(xì)胞增殖分化和脂解的影響[D];西北農(nóng)林科技大學(xué);2006年

8 吳東;高壓氧對來源于牙槽骨的成骨細(xì)胞增殖和分化的影響[D];福建醫(yī)科大學(xué);2007年

9 張琨;血管緊張素Ⅱ?qū)?T3-L1前脂肪細(xì)胞增殖及分化影響的研究[D];華中農(nóng)業(yè)大學(xué);2007年

10 吳軍兵;H-IL-6對神經(jīng)干細(xì)胞增殖、分化與遷移的作用及其分子機(jī)制[D];浙江大學(xué);2006年

，

本文編號(hào)：1968901