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REAL-TIME DETECTION OF THE EFFECTS OF ENVIRONMENTAL STRESS TO CELLULAR IMMUNITY

实时检测环境压力对细胞免疫的影响

基本信息

批准号：
13672348
负责人：
HIRANO Kazuya
金额：
$ 1.98万
依托单位：
TOKYO UNIVERSITY OF PHARMACY AND LIFE SCIENCE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672348/
关键词：
macrophage endocrine disrupting chemicals differentiation GFP 内分泌攪乱化学物質

项目摘要

This study examines the effects of environmental stress on the function of the immunological cells. First, effects of estrogenic compounds (bisphenol A, genistein and 17β-estradiol) on the ability of macrophages to produce superoxide and nitric oxide in response to stimulants was investigated. Thioglycolate-induced mouse peritoneal macrophages incubated with the estrogenic compounds were stimulated with phorbol 12-myristate 13-acetate (PMA) and lipopolysaccharide (LPS). The ability of the macrophage to produce superoxide was increased when treated with bisphenol A or genistein, but 17β-estradiol affected little. Conversely, the ability of the macrophages to produce nitric oxide in response to LPS was strongly suppressed by high concentrations of bisphenol A and genistein, 17β-estradiol was ineffective. Second; effects of bisphenol A and bisphenol A diglicidyl ether (BADGE) on the ability of monocytes to differentiate to macrophages in response to PMA and ATRA was investigated. Human mo … More nocytic leukemia THP-1 cells incubated for 7 days with bisphenol A and BADGE were stimulated with PMA and ATRA. The ability of the differentiation of THP-1 to macrophage was decreased when treated with BADGE, but bisphenol A affected little. Third; to facilitate the study of the recognition and uptake of macrophage, we developed the cell lines of THP-1 that expressed green fluorescent protein (GFP) which is a modified form of EGFP that remains bound to the plasma membrane, and that expressed cell surface recognition molecules fused to green fluorescent protein (GFP) for use in cell culture recognition models. It should be useful for a variety of cell biological studies. Finally, to establish a stable and real-time method to detect the effects of environmental stress to cellular immunity, Zeiss Axiovert 200M fluorescence microscope, CCD-camera and CO_2 incubator unit was used as a detection system. The method can be used to detect the recognition of macrophage to apoptotic cells in real time, and it can also be used to study the mechanism related to effects induced by environmental stress Less

本研究探讨了环境应激对免疫细胞功能的影响。首先，研究了雌激素化合物（双酚A、染料木黄酮和17β-雌二醇）对巨噬细胞响应刺激物产生超氧化物和一氧化氮的能力的影响。巯基乙酸诱导的小鼠腹腔巨噬细胞与雌激素化合物孵育，刺激佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）和脂多糖（LPS）。双酚A和染料木黄酮可增强巨噬细胞产生超氧阴离子的能力，而17β-雌二醇对巨噬细胞产生超氧阴离子的能力影响不大。相反，高浓度的双酚A和染料木黄酮强烈抑制巨噬细胞对LPS产生一氧化氮的能力，17β-雌二醇无效。第二个;研究了双酚A和双酚A二缩水甘油醚（BADGE）对单核细胞响应PMA和ATRA分化为巨噬细胞的能力的影响。人魔 ...更多信息用PMA和ATRA刺激用双酚A和BADGE孵育7天的黑色素白血病THP-1细胞。BADGE可降低THP-1向巨噬细胞分化的能力，而双酚A对其影响不大。第三;为了促进巨噬细胞的识别和摄取的研究，我们开发了表达绿色荧光蛋白（GFP）的THP-1细胞系，所述GFP是保持与质膜结合的EGFP的修饰形式，并且所述THP-1细胞系表达与绿色荧光蛋白（GFP）融合的细胞表面识别分子，用于细胞培养识别模型。它应该是有用的各种细胞生物学研究。最后，为了建立一种稳定、实时的检测环境应激对细胞免疫功能影响的方法，本研究采用Zeiss Axiovert 200 M荧光显微镜、CCD摄像机和CO_2培养箱作为检测系统。该方法可用于真实的实时检测巨噬细胞对凋亡细胞的识别，也可用于研究环境应激效应的相关机制。