Comparative analysis of activation induced marker (AIM) assays for sensitive identification of antigen-specific CD4 T cells
Comparative analysis of activation induced marker (AIM) assays for sensitive identification of antigen-specific CD4 T cells
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DOI：
10.1371/journal.pone.0186998
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发表时间：
2017-10-24
期刊：
影响因子：
3.7
通讯作者：
Kaufmann, Daniel E.
中科院分区：
文献类型：
--
作者：
Reiss, Samantha;Baxter, Amy E.;Kaufmann, Daniel E.
外周血和组织中抗原特异性CD4 T细胞的鉴定和研究是广泛免疫学研究的关键，包括疫苗反应和传染病。这些细胞的检测受到其稀有性和异质性的阻碍，特别是在细胞因子分泌谱方面。这些因素阻碍了通过经典方法鉴定抗原特异性CD4 T细胞的总池。我们通过测量表面激活诱导标记（AIM）的上调，开发了对此类细胞进行高灵敏度检测的检测方法。在这里，我们比较了两种基于CD69 + CD40L （CD154）或OX40 + CD25同时表达的检测方法，并开发了基于OX40 + PD-L1或4-1BB的额外AIM检测方法。我们比较了这些检测方法检测疫苗和自然感染诱导的CD4 T细胞反应的相对敏感性，并表明这些检测方法识别出不同的，但重叠的抗原特异性CD4 T细胞群，其中一个亚群也可以在细胞因子合成的基础上检测到。旁观者激活对AIM标记的影响最小。然而，一些T调节性细胞在抗原刺激下上调CD25。因此，我们验证了旨在排除大多数T调节细胞的AIM测定，用于人类和非人灵长类动物（NHP, Macaca mulatta）研究。总的来说，通过头对头比较和方法改进，我们表明AIM检测是检测抗原特异性CD4 T细胞的一种敏感和有价值的方法。
The identification and study of antigen-specific CD4 T cells, both in peripheral blood and in tissues, is key for a broad range of immunological research, including vaccine responses and infectious diseases. Detection of these cells is hampered by both their rarity and their heterogeneity, in particular with regards to cytokine secretion profiles. These factors prevent the identification of the total pool of antigen-specific CD4 T cells by classical methods. We have developed assays for the highly sensitive detection of such cells by measuring the upregulation of surface activation induced markers (AIM). Here, we compare two such assays based on concurrent expression of CD69 plus CD40L (CD154) or expression of OX40 plus CD25, and we develop additional AIM assays based on OX40 plus PD-L1 or 4-1BB. We compare the relative sensitivity of these assays for detection of vaccine and natural infection-induced CD4 T cell responses and show that these assays identify distinct, but overlapping populations of antigen-specific CD4 T cells, a subpopulation of which can also be detected on the basis of cytokine synthesis. Bystander activation had minimal effect on AIM markers. However, some T regulatory cells upregulate CD25 upon antigen stimulation. We therefore validated AIM assays designed to exclude most T regulatory cells, for both human and non-human primate (NHP, Macaca mulatta) studies. Overall, through head-to-head comparisons and methodological improvements, we show that AIM assays represent a sensitive and valuable method for the detection of antigen-specific CD4 T cells.