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

Research Project

Project/Area Number	13672348
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Environmental pharmacy
Research Institution	TOKYO UNIVERSITY OF PHARMACY AND LIFE SCIENCE
Principal Investigator	HIRANO Kazuya SHOOL OF PHARMACY, LECTURER, 薬学部, 講師 (80251221)
Co-Investigator(Kenkyū-buntansha)	BEPPU Masatoshi SHOOL OF PHARMACY,PROFESSOR, 薬学部, 教授 (60114633)
Project Period (FY)	2001 – 2002
Project Status	Completed (Fiscal Year 2002)
Budget Amount *help	¥3,100,000 (Direct Cost: ¥3,100,000) Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000) Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywords	macrophage / endocrine disrupting chemicals / differentiation / GFP / 内分泌攪乱化学物質
Research Abstract	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