The CIMT-monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8+ T lymphocytes by structural and functional assays.
The CIMT-monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8+ T lymphocytes by structural and functional assays.
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CIMT监测面板：通过结构和功能测定法对抗原特异性CD8+ T淋巴细胞进行列举的两步方法。
DOI：
10.1007/s00262-007-0378-0
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发表时间：
2008-03
影响因子：
5.8
通讯作者：
van der Burg, S. H.
中科院分区：
文献类型：
--
作者：
Britten, C. M.;Gouttefangeas, C.;Welters, M. J. P.;Pawelec, G.;Koch, S.;Ottensmeier, C.;Mander, A.;Walter, S.;Paschen, A.;Mueller-Berghaus, J.;Haas, I.;Mackensen, A.;Kollgaard, T.;thor Straten, P.;Schmitt, M.;Giannopoulos, K.;Maier, R.;Veelken, H.;Bertinetti, C.;Konur, A.;Huber, C.;Stevanovic, S.;Woelfel, T.;van der Burg, S. H.
由于可用于检测疫苗特异性T细胞应答的方法和方案多种多样，对临床试验免疫监测结果的解释是一项艰巨的任务。这种异质性以及缺乏标准导致了对公布结果的重大怀疑。因此，2005年2月，在癌症免疫治疗协会（“CIMT”）的主持下成立了一个工作组，以比较用于计数抗原特异性T细胞应答的技术和方案。在这里，我们介绍了一个国际实验室间测试项目的两个连续阶段的结果，该项目被称为“CIMT监测小组”。来自六个欧洲国家的总共13个中心参与了这项研究，其中集中制备了预测试的PBMC样品、合成肽和PE缀合的HLA四聚体并分发给参与者。所有人都被要求使用四聚体染色和一种功能测定来确定每个样品中抗原特异性T细胞的数量。第一轮测试的结果显示，分析的细胞总数是通过四聚体染色灵敏检测抗原特异性CD 8 + T细胞的最重要决定因素。ELISPOT分析受外周血单核细胞解冻后细胞数量和静息期的组合影响。因此，在第二阶段重复了这些实验，但现在要求参与者根据从第一阶段结果中提取的新指南改变他们的方案。这些建议提高了检测到的抗原特异性T细胞应答的数量，并降低了实验室之间的差异性。我们得出结论，实验室间检测的两步法允许识别影响不同T细胞检测灵敏度的不同变量，并正式表明对方案的定义校正成功地提高了灵敏度并降低了中心间变异性。这种“两步走”的实验室间项目可以为公认的国际准则确定合理的基础，从而统一用于免疫监测的技术。本文的在线版本（doi：10.1007/s 00262 -007-0378-0）包含补充材料，可供授权用户使用。
The interpretation of the results obtained from immunomonitoring of clinical trials is a difficult task due to the variety of methods and protocols available to detect vaccine-specific T-cell responses. This heterogeneity as well as the lack of standards has led to significant scepticism towards published results. In February 2005, a working group was therefore founded under the aegis of the Association for Immunotherapy of Cancer (“CIMT”) in order to compare techniques and protocols applied for the enumeration of antigen-specific T-cell responses. Here we present the results from two consecutive phases of an international inter-laboratory testing project referred to as the “CIMT monitoring panel”. A total of 13 centers from six European countries participated in the study in which pre-tested PBMC samples, synthetic peptides and PE-conjugated HLA-tetramers were prepared centrally and distributed to participants. All were asked to determine the number of antigen-specific T-cells in each sample using tetramer staining and one functional assay. The results of the first testing round revealed that the total number of cells analyzed was the most important determinant for the sensitive detection of antigen-specific CD8+ T-cells by tetramer staining. Analysis by ELISPOT was influenced by a combination of cell number and a resting phase after thawing of peripheral blood mononuclear cells. Therefore, the experiments were repeated in a second phase but now the participants were asked to change their protocols according to the new guidelines distilled from the results of the first phase. The recommendations improved the number of antigen-specific T-cell responses that were detected and decreased the variability between the laboratories. We conclude that a two-step approach in inter-laboratory testing allows the identification of distinct variables that influence the sensitivity of different T-cell assays and to formally show that a defined correction to the protocols successfully increases the sensitivity and reduces the inter-center variability. Such “two-step” inter-laboratory projects could define rational bases for accepted international guidelines and thereby lead to the harmonization of the techniques used for immune monitoring. The online version of this article (doi:10.1007/s00262-007-0378-0) contains supplementary material, which is available to authorized users.
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影响因子：
2.2
作者：
Devevre, Estelle;Romero, Pedro;Mahnke, Yolanda D.
通讯作者：
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