Whiteside T L
University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
Curr Protoc Immunol. 2001 May;Chapter 7:Unit 7.18. doi: 10.1002/0471142735.im0718s35.
In this unit, methodology is presented for measuring the capacity of human natural killer (NK) or lymphokine-activated killer (LAK) cells to lyse tumor cell targets. Cytotoxicity of these effector cells is evaluated in a short-term (51)Cr-release assay using NK-sensitive tumor cells as the targets for NK cells or NK-resistant tumor cells as the targets for LAK cells. In the basic protocol, a generic (51)Cr-release assay is described in which PBMC, purified NK cells or interleukin 2 (IL-2)-activated lymphocyte populations (LAK cells) are utilized as effector cells. Support protocols describe preparation of nonadherent tumor cells, cells obtained from malignant effusions, trypsinized tumor cells from adherent monolayer culture, or freshly isolated tumor cells from surgical specimens. All of these cell types can serve as the (51)Cr-labeled target cells in the basic protocol. The procedure for labeling target cells from any of these sources with (51)Cr is also provided.
在本单元中,介绍了用于测量人类自然杀伤(NK)细胞或淋巴因子激活的杀伤(LAK)细胞裂解肿瘤细胞靶标的能力的方法。这些效应细胞的细胞毒性在短期(51)铬释放试验中进行评估,使用对NK细胞敏感的肿瘤细胞作为NK细胞的靶标,或使用对NK细胞有抗性的肿瘤细胞作为LAK细胞的靶标。在基本方案中,描述了一种通用的(51)铬释放试验,其中外周血单核细胞(PBMC)、纯化的NK细胞或白细胞介素2(IL-2)激活的淋巴细胞群体(LAK细胞)用作效应细胞。支持方案描述了非贴壁肿瘤细胞、从恶性积液中获得的细胞、来自贴壁单层培养的胰蛋白酶消化的肿瘤细胞或来自手术标本的新鲜分离的肿瘤细胞的制备。在基本方案中,所有这些细胞类型都可以用作(51)铬标记的靶细胞。还提供了用(51)铬标记来自这些来源中任何一种的靶细胞的程序。