CD4+ and CD8+ cells in cryopreserved human PBMC maintain full functionality in cytokine ELISPOT assays
CD4+ and CD8+ cells in cryopreserved human PBMC maintain full functionality in cytokine ELISPOT assays
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DOI：
10.1016/s0022-1759(03)00226-6
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发表时间：
2003-07-01
影响因子：
2.2
通讯作者：
Tary-Lehmann, M
中科院分区：
文献类型：
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作者：
Kreher, CR;Dittrich, MT;Tary-Lehmann, M
血液中抗原特异性T细胞的频率和细胞因子特征反映了体内T细胞免疫的程度和质量。最近，对新鲜分离的外周血单核细胞（PBMC）进行的细胞因子酶联免疫斑点（ELISPOT）测定成为监测这些关键参数的有前途的工具，提供有关疫苗接种和免疫治疗疗效的直接反馈信息。然而，用新鲜分离的细胞进行ELISPOT测定在临床试验的背景下并不容易可行。获得冷冻保存样品的有效ELISPOT数据的能力将大大提高离体免疫监测能力。因此，我们系统地研究了新鲜分离的PBMC和冷冻保存后的抗原特异性T细胞反应。选择四名健康供体，其显示对六种回忆抗原的T细胞应答。将抗原反应性T细胞定义为CD 4或CD 8细胞，并通过测量干扰素（IFN）-γ、白细胞介素（IL）-2、IL-4和IL-5来确定其细胞因子效应物类别。在三个不同的时间点对供体采血，并在新鲜和冻融后测试其PBMC。结果表明，冷冻保存后，召回抗原特异性CD 4和CD 8细胞的频率和1型/2型细胞因子特征不受影响。与在人PBMC上获得的这些数据相反，鼠脾细胞的冷冻保存引起细胞因子分泌的减少。(C)2003 Elsevier B. V.保留所有权利。
The frequency and the cytokine signature of antigen-specific T cells in the blood reflect the magnitude and the quality of T cell immunity in vivo. Recently, cytokine enzyme-linked immunospot (ELISPOT) assays performed on freshly isolated peripheral blood mononuclear cells (PBMC) emerged as a promising tool for monitoring these key parameters, providing direct feedback information on the efficacy of vaccinations and immune therapies. However, performing ELISPOT assays with freshly isolated cells is not readily feasible in the context of clinical trials. The ability to obtain valid ELISPOT data on cryopreserved samples would greatly enhance ex vivo immune monitoring capabilities. We have therefore systematically studied antigen-specific T cell responses in freshly isolated PBMC and after cryopreservation. Four healthy donors were selected that displayed T cell responses to six recall antigens. The antigen reactive T cells were defined as CD4 or CD8 cells, and their cytokine effector class was established measuring interferon (IFN)-gamma, interleukin (IL)-2, IL-4 and IL-5. The donors were bled at three different time points, and their PBMC were tested fresh and after freeze-thawing. The results showed that the frequencies and type 1/type 2 cytokine signatures of recall antigen-specific CD4 and CD8 cells are unaffected after cryopreservation. in contrast to these data obtained on human PBMC, cryopreservation of murine spleen cells causes a decrease in cytokine secretion. (C) 2003 Elsevier B.V. All rights reserved.