重组蓖麻毒素B链对免疫细胞活化及信号传导通路的影响

来源 :第三届东北三省暨内蒙古自治区免疫学学术会议 | 被引量 : 0次 | 上传用户:liuyuanshuai
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  目的:探讨RTB对免疫细胞活化及细胞内的信号传导通路的影响.方法:采用CBA液相芯片检测Thl和Th2型细胞因子的分泌;流式细胞术检测小鼠CD4+和CD8+T细胞亚群数量;RT-PCR及Western blot检测iNOS基因与蛋白表达;荧光微球结合流式细胞术检测细胞吞噬功能;Elisa方法检测细胞因子分泌;蛋白质芯片检测磷酸化蛋白组学变化.结果:RTB可显著促进小鼠脾脏淋巴细胞增殖;促进淋巴细胞培养上清中Th1型细胞因子的分泌;促进CD4+T细胞的增殖和活化.RTB可促进RAW246.7细胞NO的分泌;促进iNOS基因与蛋白的表达;增强巨噬细胞的吞噬能力;促进IL-6和TNF-α基因与蛋白的表达.采用小鼠蛋白芯片抗体阵列对巨噬细胞与炎症相关的40种蛋白的变化进行检测,结果发现MCP-1、TNF-α、 MIP-1α,MIP-1β,MIP-2,RANTES,TIMP-1,CD54的表达增高,G-CSF、KC的表达下调.RTB对RAW246.7巨噬细胞活化过程中的信号转导通路的影响研究表明:酪氨酸蛋白激酶、蛋白激酶C和核因子κB信号通路可能参与RTB诱导巨噬细胞活化.提示RTB通过影响RAW246.7细胞酪氨酸蛋白激酶信号途径SRC、JAK-STAT和核因子κB信号途径IκB-α蛋白磷酸化,激活酪氨酸蛋白激酶和NF-κB信号转导途径.结论:RTB可以调节免疫细胞活化;对巨噬细胞的活化可通过细胞内的多条信号转导途径参与.本研究为RTB免疫生物学活性研究及其免疫佐剂等方面的应用奠定了新的科研依据与理论基础.
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