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作 者:许钰铃 杨婷钰 宋楠 李洁琼 XU Yuling;YANG Tingyu;SONG Nan;LI Jieqiong(Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China)
机构地区:[1]首都医科大学附属北京朝阳医院,北京100020
出 处:《标记免疫分析与临床》2024年第9期1698-1703,共6页Labeled Immunoassays and Clinical Medicine
基 金:首都医科大学附属北京朝阳医院金种子科研基金(编号:CYJZ202312)。
摘 要:目的细胞内逆向运输系统是微生物毒素蛋白导致细胞毒性的关键。KELED序列是肺炎支原体CARDS毒素逆向转运至内质网,而后发挥细胞毒性作用的关键氨基酸基序。在小鼠模型中,体外重组的CARDS蛋白可以重现肺炎支原体肺炎患者的特征性症状。然而,CARDS是否依赖于KELED序列调控肺组织炎性反应仍不清楚。方法在大肠杆菌中表达并纯化CARDS及其KELED序列突变蛋白;随后在细胞水平验证其致空泡化活性;最后,在小鼠模型中评价KELED序列突变的CARDS蛋白是否影响炎性因子的表达以及是否造成肺组织病理学损伤。结果KELED序列氨基酸发生突变,减弱CARDS蛋白致细胞空泡化效应,下调肺组织炎性因子mRNA表达,减轻肺组织的病理学损伤。结论KELED序列对CARDS蛋白致细胞空泡化及炎性损伤活性的维持至关重要。Objective Intracellular retrograde transport system is critical for cytotoxicity of microbial toxin proteins.KELED sequence is a novel key amino acid fingerprint for retrograde transport and cytotoxicity of CARDS toxin.The purified recombinant CARDS protein was administered to Balb/c mice to reproduce symptoms of human diseases caused by M.pneumoniae infections.However,it remains unclear whether CARDS relies on KELED sequences to regulate pulmonary immune responses.We aims to explore related underlying mechanism in this study.Methods CARDS and KELED mutation proteins were expressed and purified in E.coil,and then their vacuolation activities were verified with LLC cells.The Balb/c mice were then exposed to recombinant protein by intratracheal injections to evaluate its effect and function.Results KELED sequence mutation attenuated the vacuolation effect caused by CARDS protein,down-regulated the mRNA expression of inflammatory factors and alleviated the pathological damage of lung tissue.Conclusion KELED sequence is essential for the effect of vacuolation and inflammation of CARDS toxin.
关 键 词:肺炎支原体 CARDS毒素 KELED序列 炎性反应
分 类 号:R375.2[医药卫生—病原生物学]
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