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氨基葡萄糖预防自然杀伤细胞系NK-92细胞介导的细胞毒性

Glucosamine prevents cell mediated cytotoxicity of natural killer cell line NK-92
课程网址: http://videolectures.net/ipssc2017_bozic_cytotoxicity/  
主讲教师: Janja Božič
开课单位: 约瑟夫·斯特凡研究所
开课时间: 2017-05-23
课程语种: 英语
中文简介:
自然杀伤(NK)细胞是先天免疫系统中细胞毒性淋巴细胞的一个子集,在抵抗转化细胞和病毒感染细胞的早期防御中具有重要作用。在识别适当的靶细胞后,NK细胞和识别的靶细胞之间形成免疫突触,随后细胞毒性颗粒聚集在微管组织中心(MTOC)周围。细胞毒性颗粒的主要成分穿孔素和颗粒酶随后从NK细胞释放,导致靶细胞凋亡。 氨基葡萄糖是天然多糖混合物的一种成分。它被广泛用于治疗骨关节炎的疼痛,但被观察到会影响不同的细胞过程。例如,人们早就知道葡萄糖胺会影响NK细胞的活性,但其作用机制尚不清楚。为了分析细胞毒性过程,我们研究了NK-92细胞系,该细胞系对K562细胞系具有较强的细胞毒性活性。我们通过流式细胞术分析NK-92和K562细胞直接共培养的细胞毒性,使用两种标记染料DiOC18区分细胞,并使用碘化丙啶检测死亡细胞。为了激活NK-92细胞,我们使用了不同浓度的白细胞介素2(IL-2)。我们发现,细胞生长培养基中高浓度的IL-2瞬时增加ERK磷酸化。ERK磷酸化伴随着穿孔素颗粒向免疫突触的极化。为了研究氨基葡萄糖对颗粒迁移的影响,我们用不同浓度的氨基葡萄糖预处理NK-92细胞。观察到NK-92对K562细胞的细胞毒性呈剂量依赖性降低。此外,穿孔素迁移的定位显示,向培养基中添加葡萄糖胺可诱导颗粒的重定向交通,从而使细胞毒性丧失。此外,发现葡萄糖胺改变ERK磷酸化的动力学。
课程简介: Natural killer (NK) cells are a subset of cytotoxic lymphocytes of the innate immune system, which have important role in early defense against transformed and virus-infected cells. After recognition of an appropriate target cell, an immunological synapse is formed between the NK cell and the recognized target cell, followed by clustering the cytotoxic granules around the microtubule-organizing center (MTOC). The main components of cytotoxic granules, perforin and the granzymes, are then released from the NK cells, resulting in apoptosis of the target cell. Glucosamine is a component of natural polysaccharide mixtures. It is used extensively for the management of pain in osteoarthritis, but was observed to affect different cellular processes. For example, it has long been known that glucosamine effects NK cells activity, but the mechanism of action is still unknown. In order to analyze the cytotoxic process, we investigated the NK-92 cell line that possesses strong cytotoxic activity against the K562 cell line. We analyzed cytotoxicity of direct co-culture of NK-92 and K562 cells by flow cytometry, using two labelling dyes, DiOC18 to distinguish cells, and propidium iodide to detect dead cells. For activation of NK-92 cells we used different concentrations of interleukin 2 (IL-2). We showed that high concentration of IL-2 in the cell’s growth media transiently increase the ERK phosphorylation. ERK phosphorylation was accompanied by polarization of granules with perforin towards the immunological synapse. In order to study the impact of glucosamine on migration of granules, NK-92 cells were pre-treated with different concentrations of glucosamine. A dose-dependent decrease in cytotoxicity of NK-92 towards K562 cells was observed. Moreover, localization of perforin migration showed that addition of glucosamine to the medium induced redirected traffic of granules, disabling cytotoxicity towards the target cells. In addition, glucosamine was found to change dynamics of ERK phosphorylation.
关 键 词: 细胞毒性淋巴细胞; 氨基葡萄糖; 细胞毒性过程
课程来源: 视频讲座网
数据采集: 2021-11-05:zkj
最后编审: 2021-11-05:zkj
阅读次数: 75