摘要
本文采用正交设计对人外周血淋巴细胞(PBL)化学发光(CL)测定系统中的PBL、Luminol及ConA浓度进行分析比较,并探讨了新生牛血清(NCS)、牛血清白蛋白(BSA)、细胞保存温度与时间等对淋巴细胞化学发光(Ly-CL)的影响。结果表明,Ly-CL测定的最优实验条件为:(1)测定系统含1.0ml PBL悬液(1.4×10°/ml)、0.2ml Lumionl溶液(7×10^(-4)M)及0.2ml ConA溶液(700μg/ml);(2)分离的PBL悬于0.1％BSA-无酚红Hanks液中,4℃冰浴保存,2小时内完成测定。
Lymphocyte chemiluminescence (Ly-CL) is one of the early events involved in the activation of lymphocytes. This CL is thought to be result ed from the reactive oxygen species (O_2, H_2 O_2, OH, 'O_2) produced by the lymphocytes, the consequent oxidation of luminol and emission of light. In this paper we investigated the experimental condition for ConA-induced human peripheral blood lymphocyte (PBL) CL by using orthogonal design. The effects of new born calf serum (NCS), bovine serum albumin (BSA), temperature and time of cell storage on CL were also studied. The results show that the optimal condition of Ly-CL assay was 1.0 ml PBL suspension (1.4 × 10~6 cell/ml ), 0.2 ml Luminol solution ( 7 × 10^(-4)M) and 0.2 ml ConA ( 700 μg /ml ). The isolated PBL were suspended in phenol red-free Hanks solution containing 0.1% BSA and can be kept for 2 hr at 4℃ before being used without adversely affecting cell viability and CL.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
1991年第1期49-52,共4页
Chinese Journal of Immunology
关键词
外周血
淋巴细胞
化学发光
CONA
human peripheral blood lymphocyte
chemiluminescence
concanavalinA
othogonal design
