人源单核细胞系28SC体外热原检测体系的可行性研究

Feasibility study of an in vitro pyrogen detection system for human-derived monocyte cell line 28SC

关键词:  28SC  IL-6  calcitriol  LPS  LTA  酵母多糖

Key words:  28SC  IL-6  calcitriol  LPS  LTA  yeast polysaccharide

摘要:

摘要：目的 研究人源单核细胞系28SC体外检测热原的可行性。方法 用28SC模拟人体，将其分别与空白对照（IMDM）、革兰氏阴性菌（LPS）共同孵育，以酶联免疫分析（ELISA）法检测孵育体系中相关细胞因子IL-6的含量变化。活化物calcitriol对28SC孵育体系检测热原的影响，建立28SC热原检测体系；考察28SC体外检测热原体系最低检测限以及线性范围。结果及结论 活化物calcitriol浓度为500 ng·mL -1浓度时，28SC- IL-1β、TNF-α法对LPS可检测到0.5 EU·mL -1，对LTA和Zymosan无反应，28SC-IL-6法对LPS可检测到0.125 EU·mL -1，确定28SC热原检测体系的活化物calcitriol浓度为500 ng·mL -1，检测指标为细胞因子IL-6；LPS（0.0312、0.0625、0.125、0.25、0.5、1.0 EU·mL -1）刺激活化后的28SC细胞，以IL-6为检测指标，该体系对LPS的最低检测限可达到0.125 EU·mL -1，线性范围为0~0.5 EU·mL -1，R2=0.9884。

Abstract:

Abstract: OBJECTIVE To investigate the feasibility of in vitro detection of pyrogens by the human-derived monocyte cell line 28SC. METHODS 28SC were used to simulate human body, and incubated with blank control (IMDM) and Gram-negative bacteria (LPS), respectively, and changes in the content of relevant cytokine IL-6 in the incubation system were detected by enzyme-linked immunoassay (ELISA). The effect of activator calcitriol on the detection of pyrogens in the 28SC incubation system was investigated, and the 28SC pyrogens detection system was established; the lowest detection limit and the linear range of the 28SC in vitro pyrogens detection system were investigated. Results and conclusions When the concentration of activator calcitriol was 500 ng-mL -1, the 28SC- IL-1β and TNF-α assay could detect 0.5EU-mL -1 for LPS, and did not react to LTA and Zymosan, and the 28SC-IL-6 assay could detect 0.125EU-mL -1 for LPS, and the detection limit of the 28SC pyrogen assay system was determined to be 0.5EU-mL -1. system with an activator calcitriol concentration of 500 ng-mL -1 and a detection index of cytokine IL-6; LPS (0.031, 0.0625, 0.125, 0.25, 0.5, and 1.0EU-mL -1) stimulated the activated 28SC cells, and with IL-6 as the detection index, the lowest detection limit of this system for LPS could reach 0.125EU-mL -1 with a linear range of 0~0.5EU-mL -1 and R2=0.9884.