本文選題：調(diào)節(jié)性樹(shù)突狀細(xì)胞　切入點(diǎn)：內(nèi)皮型脾臟基質(zhì)細(xì)胞　出處：《內(nèi)蒙古醫(yī)學(xué)院》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：目的通過(guò)用體內(nèi)和體外兩種方法分別來(lái)誘導(dǎo)調(diào)節(jié)性樹(shù)突狀細(xì)胞的生成,同時(shí)比較了C57BL/6和NOD/LtJ兩種小鼠在誘導(dǎo)調(diào)節(jié)性樹(shù)突狀細(xì)胞能力方面的差異。方法體內(nèi)方法主要是通過(guò)用小鼠腹腔注射細(xì)菌,然后通過(guò)流式細(xì)胞術(shù)檢測(cè)脾臟內(nèi)調(diào)節(jié)性樹(shù)突狀細(xì)胞生成。首先觀察了小鼠腹腔注射細(xì)菌數(shù)在不同的數(shù)量時(shí)對(duì)調(diào)節(jié)性樹(shù)突狀細(xì)胞生成的影響;其次觀察了調(diào)節(jié)性樹(shù)突狀細(xì)胞在注射細(xì)菌后產(chǎn)生的不同的時(shí)間效應(yīng);同時(shí)也觀察了小鼠腹腔注射LPS不同劑量時(shí)對(duì)調(diào)節(jié)性樹(shù)突狀細(xì)胞的誘導(dǎo)效應(yīng);隨后又觀察了小鼠腹腔分別注射細(xì)菌和改良布氏硫乙醇酸鹽培養(yǎng)基溶液時(shí),檢測(cè)了小鼠脾臟內(nèi)調(diào)節(jié)性樹(shù)突狀細(xì)胞生成的差異;接著比較了C57BL/6和NOD/LtJ兩種小鼠注射細(xì)菌后在誘導(dǎo)調(diào)節(jié)性樹(shù)突狀細(xì)胞能力方面的差異;最后通過(guò)胞內(nèi)因子染色觀察了CD4~+Foxp3~+T細(xì)胞是否隨調(diào)節(jié)性樹(shù)突狀細(xì)胞的產(chǎn)生而產(chǎn)生。體外方法是通過(guò)把連續(xù)傳代的內(nèi)皮型脾臟基質(zhì)細(xì)胞與骨髓單個(gè)核細(xì)胞連續(xù)培養(yǎng)14天后,可使骨髓中的造血前體細(xì)胞分化成樹(shù)突樣細(xì)胞,然后通過(guò)流式細(xì)胞術(shù)對(duì)細(xì)胞表面分子進(jìn)行鑒定。結(jié)果體內(nèi)方法通過(guò)腹腔注射細(xì)菌可以誘導(dǎo)出調(diào)節(jié)性樹(shù)突狀細(xì)胞的產(chǎn)生,其中以注射細(xì)菌數(shù)為1×10~8時(shí)誘導(dǎo)效果最為明顯;而且調(diào)節(jié)性樹(shù)突狀細(xì)胞的產(chǎn)生有一定的時(shí)間效應(yīng),其在注射細(xì)菌5天后產(chǎn)生的尤為顯著;同時(shí)腹腔注射布氏硫乙醇酸鹽培養(yǎng)基溶液不會(huì)誘導(dǎo)出調(diào)節(jié)性樹(shù)突狀細(xì)胞的產(chǎn)生;而NOD/LtJ小鼠與C57BL/6小鼠相比,更易誘導(dǎo)出調(diào)節(jié)性樹(shù)突狀細(xì)胞;只是調(diào)節(jié)性樹(shù)突狀細(xì)胞不會(huì)促進(jìn)CD4~+Foxp3~+T細(xì)胞的產(chǎn)生。體外方法也同樣使骨髓中的造血前體細(xì)胞分化成了調(diào)節(jié)性樹(shù)突狀細(xì)胞。結(jié)論通過(guò)體內(nèi)和體外兩種方法均可以誘導(dǎo)出調(diào)節(jié)性樹(shù)突狀細(xì)胞。
[Abstract]:Objective to induce the formation of regulatory dendritic cells in vitro and in vivo. At the same time, the difference between C57BL / 6 and NOD/LtJ mice in inducing regulatory dendritic cells was compared. Then flow cytometry was used to detect the formation of regulatory dendritic cells in the spleen. Firstly, the effect of the number of bacteria injected into the abdominal cavity on the formation of regulatory dendritic cells was observed. The time effect of regulatory dendritic cells after bacterial injection and the induction of regulatory dendritic cells by intraperitoneal injection of LPS in mice were also observed. Then we observed the difference of regulatory dendritic cells in the spleen of mice after intraperitoneal injection of bacteria and modified brucelanolide medium respectively. Then the differences in the ability of inducing regulatory dendritic cells between C57BL / 6 and NOD/LtJ mice were compared. Finally, the intracellular factor staining was used to observe whether CD4 ~ Foxp3~ T cells were produced by regulatory dendritic cells. In vitro, the endothelial splenic stromal cells and bone marrow mononuclear cells were cultured continuously for 14 days. The hematopoietic precursor cells in bone marrow were differentiated into dendritic cells and then identified by flow cytometry. Results the regulatory dendritic cells could be induced by intraperitoneal injection of bacteria in vivo. Among them, the number of injected bacteria was 1 脳 10 ~ 8:00, the induction effect was the most obvious, and the production of regulatory dendritic cells had a certain time effect, especially after 5 days of injection. At the same time, the regulatory dendritic cells were not induced by intraperitoneal injection of bruclear thioglycolate medium solution, but the regulatory dendritic cells were more easily induced in NOD/LtJ mice than in C57BL / 6 mice. Only regulatory dendritic cells did not promote the production of CD4 ~ Foxp3 ~ T cells. In vitro, the hematopoietic precursor cells in bone marrow were also differentiated into regulatory dendritic cells. Conclusion both in vivo and in vitro methods can be used to produce regulatory dendritic cells. Regulatory dendritic cells were induced.
【學(xué)位授予單位】：內(nèi)蒙古醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R392.1

【共引文獻(xiàn)】

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本文編號(hào)：1579494