Frequency and reactivity of antigen-specific T cells were concurrently measured through the combination of artificial antigen-presenting cell, MACS and ELISPOT.
Frequency and reactivity of antigen-specific T cells were concurrently measured through the combination of artificial antigen-presenting cell, MACS and ELISPOT.
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通过人工抗原呈递细胞、MACS和ELISPOT的组合，同时测量抗原特异性T细胞的频率和反应性。
DOI：
10.1038/s41598-017-16549-1
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发表时间：
2017-11-27
影响因子：
4.6
通讯作者：
Qiu J
中科院分区：
文献类型：
--
作者：
Shen C;Xu T;Wu Y;Li X;Xia L;Wang W;Shahzad KA;Zhang L;Wan X;Qiu J
传统的肽-主要组织相容性复合物（pMHC）多聚体染色，细胞内细胞因子染色和酶联免疫斑点（ELISPOT）测定不能同时确定抗原特异性T细胞（AST）的频率和反应性在一个单一的测定。在这份报告中，pMHC多聚体，磁激活细胞分选（MACS），ELISPOT技术已被集成到一个微孔耦合pMHC多聚体上的细胞大小的磁珠，以表征AST细胞群在96孔微孔板，预先包被的精氨酸捕获抗体。该方法无需流式细胞仪或荧光强度扫描，可在一次检测中对AST细胞进行计数和局部细胞因子的产生，具有广泛的应用前景。在此，测量来自OT-1 T细胞受体（TCR）转基因小鼠的卵清蛋白257 -264特异性CD 8 + T细胞。计数AST细胞的方法学准确性、特异性、可重复性和灵敏度与常规pMHC多聚体染色相比良好。同时，应用AAPC-微孔板检测患者外周血B病毒（HBV）核心抗原18 -27和表面抗原183 -191特异性CD 8 + T细胞的频率和反应性，并与常规方法进行比较。该方法不需要高端仪器，可便于转化研究中对给定抗原的患者特异性细胞免疫应答模式的常规分析。
Conventional peptide-major histocompatibility complex (pMHC) multimer staining, intracellular cytokine staining, and enzyme-linked immunospot (ELISPOT) assay cannot concurrently determine the frequency and reactivity of antigen-specific T cells (AST) in a single assay. In this report, pMHC multimer, magnetic-activated cell sorting (MACS), and ELISPOT techniques have been integrated into a micro well by coupling pMHC multimers onto cell-sized magnetic beads to characterize AST cell populations in a 96-well microplate which pre-coated with cytokine-capture antibodies. This method, termed AAPC-microplate, allows the enumeration and local cytokine production of AST cells in a single assay without using flow cytometry or fluorescence intensity scanning, thus will be widely applicable. Here, ovalbumin257–264-specific CD8+ T cells from OT-1 T cell receptor (TCR) transgenic mice were measured. The methodological accuracy, specificity, reproducibility, and sensitivity in enumerating AST cells compared well with conventional pMHC multimer staining. Furthermore, the AAPC-microplate was applied to detect the frequency and reactivity of Hepatitis B virus (HBV) core antigen18–27- and surface antigen183–191-specific CD8+ T cells for the patients, and was compared with conventional method. This method without the need of high-end instruments may facilitate the routine analysis of patient-specific cellular immune response pattern to a given antigen in translational studies.
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影响因子：
15.8
作者：
Chen, DS;Soen, Y;Stuge, TB;Lee, PP;Weber, JS;Brown, PO;Davis, MM
通讯作者：
Davis, MM
影响因子：
1.2
作者：
Axmann, Markus;Schuetz, Gerhard J.;Huppa, Johannes B.
通讯作者：
Huppa, Johannes B.
DOI：
10.1073/pnas.0407019102
发表时间：
2005-03-08
影响因子：
11.1
作者：
Stone, JD;Demkowicz, WE;Stern, LJ
通讯作者：
Stern, LJ
影响因子：
4.4
作者：
Yue, C.;Oelke, M.;Schneck, J. P.
通讯作者：
Schneck, J. P.
影响因子：
56.9
作者：
Altman, JD;Moss, PAH;Davis, MM
通讯作者：
Davis, MM