体外诱导生成树突状细胞的成熟度和激活性研究.docVIP

体外诱导生成树突状细胞的成熟度和激活性研究 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：体外诱导生成树突状细胞的成熟度和激活性研究 2 1 材料与方法 3 12 方法 3 121 外周血单核细胞（PBM）的分离 3 122 DC的培养 3 123 细胞形态学观察 3 124 细胞表型测定 4 125 统计学方法 4 2 结果 4 21 培养细胞的形态学 4 22 培养细胞的免疫表型 5 3 讨论 5 文2：内毒素激活巨噬细胞的信号传导 6 1 LBP的结构和功能 7 2 CD14介导细胞激活的信号传导 8 3 内毒素激活巨噬细胞的胞内信号传导 9 4 与内毒素激活细胞有关的转录因子和反应元件 10 5 结语 11 参考文摘引言： 11 原创性声明（模板） 12 文章致谢（模板） 13 正文 体外诱导生成树突状细胞的成熟度和激活性研究 文1：体外诱导生成树突状细胞的成熟度和激活性研究 【ABSTRACT】 Objective: To study maturity and activation of dendritic cells (DC) induced from human peripheral blood monocytes（PBM）. Methods: The monocytes collected from nomal peripheral blood were cultured with GMCSF (1000U/ml) and IL4 (500U/ml) for 7 days, then cultured with TNFα (100ng/ml) for 3 days. MHCⅡand costimulatory molecules of DC were analyzed by flow cytometry for two times, so maturity　and activation of DC were known by them. Results: CD83、CD86、CD11c、CD64、HLADR expressio in PBM were 226%、555％、361％、32％、134％ after PBM were cultured with GM-CSF and IL4 for 7 days, but their expressio in PBM were 815%、993%、988％、34％、833％ after PBM were cultured with TNFαfor 3 days. Conclusion: DC induced from PBM with GMCSF and IL4 for 7days were not mature, but the system inducement was beneficial to expaion of DC. Cultured with TNFαfor 3 days , DC induced form PBM were very good in maturity and activation. 【KEY WORDS】 Dendritic cells; Culture in vitro; Maturity; Activation 树突状细胞（dendritic cell,DC）是体内最强的抗原提呈细胞 ，免疫反应的产生首先是由抗原提呈细胞 （APC）捕获抗原，经其加工处理后将抗原信号传递给T、B淋巴细胞，从而引发一系列的免疫应答。因此，APC是机体免疫反应的首要环节，能否进行有效的抗原呈递直接关系到免疫激活和免疫耐受的诱导。成熟的DC启动免疫应答，而不成熟的DC参与免疫耐受的形成。DC通常从骨髓CD34+细胞和PBM培养获得。在此，我们探索了从PBM培养DC的成熟度和激活性的方法。 1 材料与方法 11 材料 淋巴细胞分离液购自Sigma公司，丝裂酶素C购自Sigma公司，RPMI1640购自GIBCO公司，胎牛血清购自GIBCO公司，rhGMCSF、rhIL4、hTNFa购自RD公司，CD83、CD11c、CD64、CD86、HLADR、FITC标记兔抗鼠IgG荧光二抗购自华美生物工程公司。 12 方法 121 外周血单核细胞（PBM）的分离 采集健康人外周血100ml，肝素抗凝，用淋巴细胞分离液分离出单个核细胞，RPMI1640悬浮后种于24孔板，置于37℃，5%的CO2培养箱中，培养3h，弃去悬浮细胞，贴壁细胞即为PBM，用于DC培养。 122 DC的培养 含10%胎牛血清的RBMI