Sepsis is systemic inflammatory response syndrome(SIRS)associated with the presence of pathogenic microorganisms or their ***(LPS)is an important component of the outer membrane of Gram negative bacteria and has a piv...
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Sepsis is systemic inflammatory response syndrome(SIRS)associated with the presence of pathogenic microorganisms or their ***(LPS)is an important component of the outer membrane of Gram negative bacteria and has a pivotal role in inducing Gram negative *** play an essential role in infection and *** recognition of LPS is of particular importance in the defense system and pattern recognition receptors(PRR)have been considered to be important in initial steps for cellular recognition of LPS and consequence initiation of LPS *** the past few years,intense research in the fields of PRR and their recognition mechanism has been *** this review,we attempt to expound recent research advances of macrophage PRRs for LPS recognition and their mechanism.
目的:通过对影响细胞ELISA的关键条件和步骤进行优化和改进,以降低实验假阳性率,提高敏感性。方法:THP-1细胞接种96孔培养板,分别用脂多糖(LPS)刺激或不刺激作为处理组和对照组,将通过噬菌体展示技术筛选获得的噬菌体分别与处理组及对照组细胞作用,洗脱未结合的噬菌体,加入HRP标记的抗M13单克隆抗体,分别用酶标仪和Kodak IS 2000R数码成像系统检测细胞与短肽的结合情况。并针对细胞的特性,在细胞的准备、固定、孵育和洗涤等关键步骤进行改进。结果:酶标仪和数码成像系统检测分别得到6个和8个可与细胞高亲和力结合的阳性噬菌体克隆,经过免疫荧光实验证实数码成像系统检测得到的8个阳性克隆均与细胞结合,并且数码成像系统多次测量值具有很好的重复性。结论:本方法为蛋白质与完整细胞的亲和活性研究提供了有效的实验方案,具有较好的应用前景。
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