公开(公告)号：US20070224660A1

公开(公告)日：2007-09-27

申请号：US11578477

申请日：2005-04-14

申请人： Yan Cui ,  Kong Chen

发明人： Yan Cui ,  Kong Chen

IPC分类号： C12Q1/06

CPC分类号： C12N5/0636 ,  C12N2503/00 ,  G01N33/5005 ,  G01N33/505

摘要： A new cytotoxic T lymphocyte (CTL) assay has been discovered using two cell lines that stably express either green fluorescent protein (GFP) or red fluorescent protein (DsRed), which are distinguishable by FACS, fluorescence microplate reader, or fluorescence microscopy. Using one cell line as a target (T) to present antigen and the other, at the same number, as an internal control (reference, R), a new CTL assay (named fluorolysometric (FL)-CTL assay) way developed based on cytolysis of these fluorescent protein-expressing targets detectable by FACS. This FL-CTL assay was further extended for use with a fluorescent microplate reader. This FL-CTL assay was reproducibly used to determine primary CTL activity at high sensitivity when compared to other conventional assays with in vivo activated T cells against different antigens. This new reliable, sensitive, convenient, and economical CTL assay has broad application potentials for experimental and clinical use in different antigen and effector-target systems.

摘要翻译： 已经发现使用稳定表达绿色荧光蛋白（GFP）或红色荧光蛋白（DsRed）的两种细胞系的新的细胞毒性T淋巴细胞（CTL）测定法，其可通过FACS，荧光酶标仪或荧光显微镜鉴别。 使用一种细胞系作为靶标（T），以相同的数量，作为内部对照（参考文献R），以新的CTL测定（称为荧光分析（FL）-CTL测定）方式，基于 通过FACS检测的这些荧光蛋白表达靶点的细胞溶解。 该FL-CTL测定进一步扩展以用于荧光酶标仪。 与其他针对不同抗原的体内激活的T细胞的常规测定相比，该FL-CTL测定可重复地用于确定高灵敏度的初级CTL活性。 这种新的可靠，灵敏，方便和经济的CTL测定在不同的抗原和效应靶系统中具有广泛的实验和临床应用潜力。