ELISPOT assay to measure antigen-specific murine CD8+ T cell responses
ELISPOT assay to measure antigen-specific murine CD8+ T cell responses
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DOI：
10.1016/s0022-1759(01)00331-3
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发表时间：
2001-06-01
影响因子：
2.2
通讯作者：
Zavala, F
中科院分区：
文献类型：
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作者：
Carvalho, LH;Hafalla, JCR;Zavala, F
酶联免疫斑点技术（ELISPOT）依赖于在用抗原进行体内刺激后单个T细胞的细胞因子分泌的可视化。该测定法已被开发和标准化，用于定量检测经受不同免疫方案的小鼠中的抗原特异性CD 8（+）T细胞[J. Immunol. Methods 181（1995）45]。我们已经确定了影响ELISPOT测定效力的重要变量，在本方案中，我们详细描述了这种方法。作为模型，我们使用了表达H-2K（d）限制性SYVPSAEQI的疟疾子孢子免疫的小鼠外周血、脾脏和肝脏中的CDS' T细胞产生干扰素γ。该方案也已成功地用于检测Th 1和Th 2表位特异性CD 4 + T细胞。(C)2001 Elsevier Science B. V.保留所有权利。
The enzyme-linked immunospot technique (ELISPOT) relies on the visualization of cytokine secretion by individual T cells following in vivo stimulation with antigen. This assay has bren developed and standardized for the quantitative detection of antigen-specific CD8(+) T cells in mice subjected to different immunization protocols [J. Immunol. Methods 181 (1995) 45]. We have identified important variables that affect the efficacy of the ELISPOT assay and in this protocol we describe this methodology in detail. As a model, we used the production of interferon-gamma by CDS' T cells from peripheral blood, spleen and Liver of mice immunized with malaria sporozoites expressing the H-2K(d)-restricted SYVPSAEQI. This protocol has also been used successfully to detect Th1 and Th2 epitope specific CD4+ T cells. (C) 2001 Elsevier Science B.V. All rights reserved.