一种纯化绵羊红细胞玫瑰花结淋巴细胞的新方法。

A novel method for the purification of sheep red cell rosetting lymphocytes.

作者信息

Mikamo S

机构信息

Department of Clinical Oncology, University of Tokyo, Japan.

出版信息

J Immunol Methods. 1988 Mar 16;107(2):189-96. doi: 10.1016/0022-1759(88)90217-7.

DOI:10.1016/0022-1759(88)90217-7

PMID:3346550

Abstract

A rapid, direct method for the purification of sheep red cell rosetting lymphocytes (ERFC) was developed. The whole procedure, including rosette formation, density separation and hemolysis could be completed within 10 min. A mixture of human peripheral blood mononuclear cells (PBMC) and 2-aminoethylisothiouronium bromide hydrobromide-treated sheep erythrocytes (EAET) was layered onto Ficoll-Paque without any pretreatment and centrifuged at 600 X g for 2.5 min. The pellet was then immediately treated with an NH4Cl solution containing 10% FCS and hemolysis was completed within 1 min. The purity of ERFC separated in one cycle of the procedure was 98%, the viability 99% and the yield 56% of the initial lymphocyte count. The re-rosetting ability of the prepared cells, after hemolysis, was 95%. The lymphocytes in the fraction prepared by the same method contained 94.3% CD2(OKT11)+ cells, 90% of which were CD3(OKT3)+ cells (T cells) and 9% were CD16(Leu11a)+ cells (NK cells).

摘要

开发了一种快速、直接的纯化绵羊红细胞玫瑰花结形成淋巴细胞（ERFC）的方法。整个过程，包括玫瑰花结形成、密度分离和溶血，可在10分钟内完成。将人外周血单个核细胞（PBMC）和经氢溴酸2-氨基乙基异硫脲处理的绵羊红细胞（EAET）的混合物无需任何预处理就铺在Ficoll-Paque上，并以600×g离心2.5分钟。然后立即用含有10%胎牛血清的氯化铵溶液处理沉淀，溶血在1分钟内完成。该方法一个循环中分离出的ERFC纯度为98%，活力为99%，产量为初始淋巴细胞计数的56%。溶血后制备的细胞的再玫瑰花结形成能力为95%。用相同方法制备的组分中的淋巴细胞含有94.3%的CD2(OKT11)+细胞，其中90%是CD3(OKT3)+细胞（T细胞），9%是CD16(Leu11a)+细胞（NK细胞）。