Santhi G, Ramanadham M
Department of Biochemistry, University of Hyderabad, India.
Indian J Exp Biol. 1995 Dec;33(12):926-30.
A modified procedure for isolation of intraepithelial lymphocytes (IEL) from rat small intestine was developed which makes use of a protease inhibitor-phenyl methyl sulfonyl fluoride (PMSF) in the isolation medium. Yield and viability of IEL isolated in presence of PMSF were significantly higher as compared to those isolated in absence of PMSF. IEL isolated in presence PMSF demonstrated a significantly higher proliferative response to concanavalin A (con A) compared to those isolated in absence of PMSF. The natural killer cell activity of IEL population isolated in presence of PMSF using YAC-1 lymphoma cells as targets was also significantly higher compared to those isolated in absence of PMSF. Phenotypic analysis using monoclonal antibodies specific for rat T lymphocyte subsets revealed that majority of IEL were cytotoxic/suppressor phenotype with a minor subset of helper/inducer phenotype. The method described yielded a population of IEL which is suitable for further functional studies in vitro.
开发了一种从大鼠小肠中分离上皮内淋巴细胞（IEL）的改良方法，该方法在分离培养基中使用了蛋白酶抑制剂——苯甲基磺酰氟（PMSF）。与未使用PMSF分离的IEL相比，在PMSF存在下分离的IEL的产量和活力显著更高。与未使用PMSF分离的IEL相比，在PMSF存在下分离的IEL对刀豆球蛋白A（Con A）表现出显著更高的增殖反应。以YAC-1淋巴瘤细胞为靶标，在PMSF存在下分离的IEL群体的自然杀伤细胞活性也显著高于未使用PMSF分离的IEL。使用针对大鼠T淋巴细胞亚群的单克隆抗体进行的表型分析显示，大多数IEL为细胞毒性/抑制表型，一小部分为辅助/诱导表型。所描述的方法产生了适合于进一步体外功能研究的IEL群体。
