發(fā)布時(shí)間：2018-06-02 11:53

  本文選題：新金分枝桿菌 + 調(diào)節(jié)性T細(xì)胞�。� 參考：《吉林農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：近年來(lái),臨床上感染非結(jié)核分枝桿菌的病例不斷增加,非結(jié)核分枝桿菌病已經(jīng)成為危害人類(lèi)生命和健康的重要問(wèn)題。目前尚無(wú)治療非結(jié)核分枝桿菌病的特效藥。調(diào)節(jié)性T細(xì)胞是負(fù)向調(diào)節(jié)機(jī)體免疫反應(yīng)的一類(lèi)T細(xì)胞亞群,它既能夠抑制機(jī)體不恰當(dāng)?shù)拿庖叻磻?yīng),還能限定免疫應(yīng)答的范圍和程度。因此本試驗(yàn)通過(guò)對(duì)感染新金分枝桿菌的C57BL/6小鼠進(jìn)行研究,分析其感染期間對(duì)Tregs亞群表型及功能的影響,使我們對(duì)非結(jié)核分枝桿菌感染期間機(jī)體Tregs亞群免疫機(jī)制有一個(gè)新的認(rèn)識(shí)。本試驗(yàn)選取雌性C57BL/6小鼠,經(jīng)腹腔注射感染新金分枝桿菌。分別于感染后1、2、3、4、5、6、7、9、11、13、16、18、23周采用流式細(xì)胞術(shù)檢測(cè)脾臟CD_4~+CD_(25)~+Fox P3~+Treg與CD8~+CD_(25)~+Fox P3~+Treg比例變化;并用CD152檢測(cè)表型變化明顯的時(shí)間點(diǎn)檢測(cè)其抑制性功能;利用免疫磁珠分選CD_4~+CD_(25)~+Treg與CD_4~+CD_(25)-T細(xì)胞進(jìn)行MTT增殖抑制試驗(yàn);通過(guò)熒光定量PCR技術(shù)檢測(cè)CD_4~+CD_(25)~+Treg中Fox P3、TGF-β、IL-10和IFN-γ4種細(xì)胞因子m RNA水平表達(dá)量的變化。結(jié)果顯示:1、新金分枝桿菌感染的小鼠脾臟中,Tregs亞群主要以CD_4~+CD_(25)~+FoxP3~+Treg表型存在,并在第6周其所占細(xì)胞比例達(dá)到最高,約占2%。CD_4~+CD_(25)~+CD152~+細(xì)胞與CD_4~+CD_(25)~+Fox P3~+細(xì)胞比例呈正相關(guān)。2、利用免疫磁珠分選的CD_4~+CD_(25)-T細(xì)胞及CD_4~+CD_(25)~+Treg細(xì)胞純度較高,CD_4~+CD_(25)-T細(xì)胞為82.5%,CD_4~+CD_(25)~+Treg細(xì)胞可以達(dá)到79.1%。CD_4~+CD_(25)~+Treg對(duì)CD_4~+CD_(25)-T細(xì)胞產(chǎn)生抑制性作用,CD_4~+CD_(25)-T細(xì)胞的抑制率為89.8%。3、CD_4~+CD_(25)~+Treg分泌的FoxP3、IL-10與TGF-β的mRNA相對(duì)表達(dá)量均與對(duì)照組差異顯著,IFN-γm RNA表達(dá)量與對(duì)照組無(wú)統(tǒng)計(jì)學(xué)差異。綜上,新金分枝桿菌感染小鼠脾臟Tregs亞群主要以CD_4~+CD_(25)~+Fox P3存在,發(fā)揮其免疫抑制作用。體外增殖抑制試驗(yàn)中,CD_4~+CD_(25)~+Treg能夠抑制CD_4~+CD_(25)-T細(xì)胞的增殖作用,驗(yàn)證了CD_4~+CD_(25)~+Treg能夠通過(guò)直接接觸發(fā)揮抑制性功能。CD_4~+CD_(25)~+Treg細(xì)胞主要通過(guò)分泌細(xì)胞因子TGF-β與IL-10來(lái)發(fā)揮其免疫抑制作用。
[Abstract]:In recent years, the number of patients infected with non-tuberculous mycobacterium has been increasing, and non-tuberculosis mycoidosis has become an important problem that endangers human life and health. At present, there is no specific drug for the treatment of non-tuberculosis mycobacterium disease. Regulatory T cells are a subgroup of T cells that negatively regulate the immune response of the body. It can not only inhibit the inappropriate immune response, but also limit the scope and degree of immune response. Therefore, through the study of C57BL/6 mice infected with Mycobacterium neoformans, the effects of the infection on the phenotype and function of Tregs subgroup were analyzed, so that we could have a new understanding of the immune mechanism of Tregs subgroup during the period of non-tuberculosis mycobacterium infection. In this study, female C57BL/6 mice were injected intraperitoneally with Mycobacterium neoformans. The changes of CD4 ~ Fox P3 ~ Treg in spleen and CD8 ~ CDT 25 ~ Fox P3 ~ Treg in spleen were detected by flow cytometry at week 1, 2, 2, 2, 3, 5, 5, 5, 9, 11, 13, 16, 18, 23 weeks after infection, and the inhibitory function was detected by CD152 at a time point when phenotypic changes were obvious. The expression of Fox P3TGF- 尾 IL-10 and IFN- 緯 in CD4 ~ CDGF-尾 -IL-10 and IFN- 緯 were detected by fluorescence quantitative PCR technique. The results showed that Tregs subsets in the spleen of mice infected with Mycobacterium neoformans mainly existed in the phenotype of CD4 ~ CDSP-25 ~ FoxP3 ~ Treg, and reached the highest percentage of cells in the 6th week. There was a positive correlation between the proportion of CD4- Fox P3~ cells and the proportion of CD4- Fox P3~ cells. The purity of CD4- CD_(25)-T cells and CD4- Treg cells was 82.5 CD4CD4- CD152- CD_(25)-T cells. The CD4- CD_(25)-T cells could reach 79.1. CD4CD4- Fox P3~ + cells. The CD4- CD_(25)-T cells could inhibit the CD4- CD_(25)-T cells in the cells of 79.1. CD4CD4CDT / Treg. The CD4- CD_(25)-T cells of the population could reach 79.1.CD4CD4CDT / Treg could inhibit the proliferation of CD4- CD_(25)-T cells by using immunomagnetic beads to separate the CD4- CD_(25)-T cells and the CD4- Treg cells with the purity of CD4- CD_(25)-T cells. The inhibitory rate of CD4 ~ CD_(25)-T cells was 89.8.3. there was no significant difference in the relative expression of mRNA between the two groups in the expression of FoxP3- 緯 m RNA and TGF- 尾 secreted by 25 ~ Treg of CD4 ~ CDD. There was no significant difference in the expression of IFN- 緯 m RNA between the two groups. In conclusion, the spleen Tregs subsets of mice infected with Mycobacterium neoformans were mainly CD4 ~ CDSP-25 ~ Fox P3, which played an immunosuppressive effect. In vitro proliferation inhibition test, CD4 ~ CDS ~ Treg can inhibit the proliferation of CD4- CD_(25)-T cells, which proves that CD4 ~ CDS-1 ~ Treg can exert inhibitory function by direct contact. CD4- 尾 and IL-10 can play its immunosuppressive effect mainly by secreting cytokines TGF- 尾 and IL-10.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R52

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 汪煉超;李莉;;肺結(jié)核患者血清與痰上清液中TNF-α、IL-4、IFN-γ、TGF-β的表達(dá)及意義[J];實(shí)用臨床醫(yī)藥雜志;2016年09期

2 陳曉紅;吳迪;;肺外非結(jié)核分枝桿菌病的臨床特點(diǎn)和診斷[J];中國(guó)醫(yī)刊;2016年03期

3 唐神結(jié);;非結(jié)核分枝桿菌病診斷與治療專(zhuān)家共識(shí)解讀[J];中國(guó)醫(yī)刊;2016年03期

4 張哲峰;梁華剛;薛雷;杜海榮;張璐;賈瑞;高鯤;;重癥肌無(wú)力患者外周血Foxp3和CD152表達(dá)變化及其相關(guān)性分析[J];中風(fēng)與神經(jīng)疾病雜志;2016年02期

5 李紅;;非結(jié)核分枝桿菌肺病患者血清細(xì)胞因子測(cè)定臨床意義分析[J];當(dāng)代醫(yī)學(xué);2016年06期

6 莊戰(zhàn)強(qiáng);許文華;吳元波;張翠萍;李慶;任明山;;糖皮質(zhì)激素對(duì)重癥肌無(wú)力患者外周血調(diào)節(jié)性T細(xì)胞中Foxp3及其胞內(nèi)CTLA-4表達(dá)的影響[J];安徽醫(yī)科大學(xué)學(xué)報(bào);2015年09期

7 齊花蕊;邱佳熠;王春芳;錢(qián)愛(ài)東;;調(diào)節(jié)性T細(xì)胞及在結(jié)核分支桿菌感染中的作用研究進(jìn)展[J];動(dòng)物醫(yī)學(xué)進(jìn)展;2014年12期

8 陳興年;葉志堅(jiān);吳妹英;;非結(jié)核分枝桿菌肺病血清中細(xì)胞因子的檢測(cè)及臨床意義[J];臨床肺科雜志;2014年03期

9 張志強(qiáng);賀杰峰;張瑜;蘭志偉;趙浩亮;;大鼠肝癌免疫微環(huán)境中Treg與IL-10水平的變化及意義[J];當(dāng)代醫(yī)學(xué);2014年04期

10 蒯守剛;戴亞萍;何琳靜;王旭;劉君;劉斌;裴豪;;IL-17、IL-1β和IL-10在結(jié)核病患者中的表達(dá)及臨床意義[J];檢驗(yàn)醫(yī)學(xué);2012年11期

相關(guān)碩士學(xué)位論文 前2條

1 岳麗敏;非結(jié)核分枝桿菌對(duì)小鼠CD4~+T細(xì)胞分化影響的研究[D];吉林農(nóng)業(yè)大學(xué);2015年

2 陳書(shū)平;Cd8~+Cd25~+FOXP3~+調(diào)節(jié)性T淋巴細(xì)胞在惡性胸腔積液中的表達(dá)及其臨床意義[D];廣西醫(yī)科大學(xué);2011年

，

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