Enumerating antigen-specific T-Cell responses in peripheral blood - A comparison of peptide MHC tetramer, ELISpot, and intracellular cytokine analysis
Enumerating antigen-specific T-Cell responses in peripheral blood - A comparison of peptide MHC tetramer, ELISpot, and intracellular cytokine analysis
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DOI：
10.1097/00002371-200501000-00008
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发表时间：
2005-01-01
影响因子：
3.9
通讯作者：
Clay, TM
中科院分区：
文献类型：
--
作者：
Hobeika, AC;Morse, MA;Clay, TM
检测循环抗原特异性T细胞对免疫的反应是癌症疫苗临床试验的重要生物学终点。通常使用的分析方法是多肽MHC四聚体、ELISpot和细胞内细胞因子分析。尽管在这些检测中对阳性反应的定义没有达成一致意见，但许多小组已经选择了一些T细胞，这些T细胞的数量比阴性对照的平均值高出2个标准差。作者希望确定在检测模型抗原(巨细胞病毒(CMV)pp65的免疫优势抗原-A*0201限制性表位)的阳性和阴性T细胞反应方面，这一分界点在每一种检测中的表现如何。对于每一项检测，CMV血清阴性反应的平均+2标准差是最好区分两组的点。使用该值，每种检测方法的敏感性为87.5%，特异性为95%-100%，与其他两种方法显示出高度的一致性(kappa 0.76-0.9)。作者总结说，目前可用的免疫学检测方法在检测与生物相关的抗原特异性T细胞水平方面表现良好。这些检测将更好地确定针对病毒疾病的保护性免疫反应的数量和质量，并提供对保护性抗癌免疫要求的洞察。
Detection of the circulating antigen-specific T-cell response to immunization is an important biologic end point in clinical trials of cancer vaccines. Typically employed assays are peptide MHC tetramer, ELISpot, and intracellular cytokine analysis. Although there is no agreement on the definition of a positive response in these assays, many groups have chosen a number of T cells greater than 2 standard deviations above the mean of the negative controls. The authors wished to determine how well this cutoff performed for each of these assays in detecting positive and negative T-cell responses to a model antigen, the immunodominant HLA-A*0201-restricted epitope of cytomegalovirus (CMV) pp65. For each assay, the mean + 2 standard deviations of the response for CMV seronegatives was the point that best separated the two groups. Using this value, each assay had a sensitivity of 87.5% and specificity of 95% to 100% and exhibited a high degree of concordance (kappa 0.76-0.9) with the other two. The authors conclude that currently available immunologic assays perform well in detecting biologically relevant levels of antigen-specific T cells. These assays will better define the quantity and quality of protective immune responses to viral disease and offer insight into the requirements for protective anti-cancer immunity.