• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Clinical immunology: The official journal of the Clinical Immunology Society >Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8(+) T cells and MHC class I expressing antigen presenting cell lines
【24h】

Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8(+) T cells and MHC class I expressing antigen presenting cell lines

机译：ELISpot检测方法的可变性：抗原反应性细胞的特异性检测需要纯化的CD8（+）T细胞和表达MHC I类的抗原呈递细胞系

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Quantification of antigen-specific CD8(+) T cells is important for monitoring infection, vaccination, and response to therapy in cancer and immune-mediated diseases. Cytokine enzyme-linked-immunospot (ELISpot) assays are often used for this purpose. We found that substantial spot formation in IFN gamma ELISpot assays occurred independently of CD8(+) T cells even when classical MHC class I restricted peptides are used for stimulation. Using fractionated cells and intracellular cytokine staining, the non-CD8(+) T cell IFN-gamma production was attributed to the CD4(+) T cell fraction. We therefore refined a cell line-based ELISpot assay combining HLA-A*0201 expressing K562 cells for antigen presentation with purified CD8(+) T cells and demonstrated that it specifically detected CD8(+) T cell responses with detection limits comparable to traditional ELISpot assays and dextramer-based quantification. The assay was further adapted to whole antigen responses with antigen (pre-proinsulin)-expressing HLA-A*0201K562 cells. Thus, we revealed and corrected a weak spot of the CD8(+) ELISpot assay. (C) 2015 Elsevier Inc. All rights reserved.
机译：抗原特异性CD8（+）T细胞的定量对于监测感染，疫苗接种以及对癌症和免疫介导疾病的治疗反应至关重要。细胞因子酶联免疫斑点法（ELISpot）通常用于此目的。我们发现，即使经典的MHC I类限制性肽用于刺激，IFNγELISpot分析中的大量斑点形成也独立于CD8（+）T细胞而发生。使用分级细胞和细胞内细胞因子染色，非CD8（+）T细胞IFN-γ产生归因于CD4（+）T细胞部分。因此，我们改进了基于细胞系的ELISpot检测方法，将表达HLA-A * 0201的K562细胞用于抗原呈递与纯化的CD8（+）T细胞相结合，并证明了它可以特异性检测CD8（+）T细胞反应，且检测限可与传统ELISpot媲美分析和基于右旋体的定量。该测定进一步适用于表达抗原（胰岛素原前）的HLA-A * 0201K562细胞的全抗原反应。因此，我们揭示并纠正了CD8（+）ELISpot分析的弱点。 （C）2015 Elsevier Inc.保留所有权利。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号