Tassignon Joël, Burny Wivine, Dahmani Samira, Zhou Lin, Stordeur Patrick, Byl Baudouin, De Groote Donat
CYPRO SA, Brussels, Belgium.
J Immunol Methods. 2005 Oct 30;305(2):188-98. doi: 10.1016/j.jim.2005.07.014. Epub 2005 Aug 31.
One of the challenges for immunomonitoring in clinical trials is to detect an antigen specific T cell-mediated immune response. In an attempt to define the most suitable assay, tetanus toxoid was used to compare the capacity of 4 different methods to detect cytokine responses, before and after recall vaccination, in peripheral blood mononuclear cells (PBMC) of 14 healthy volunteers. ELISA, ELISPOT, intracytoplasmic detection and real-time RT-PCR were chosen to measure IFN-gamma production before and after vaccination. As far as the detection of memory T cell status (before vaccination) was concerned, we found that ELISPOT was the most sensitive method to discriminate TT-induced from spontaneous responses. On the other hand, intracytoplasmic cytokine detection was the most efficient method to detect the restimulating effect of TT vaccination.
临床试验中免疫监测面临的挑战之一是检测抗原特异性T细胞介导的免疫反应。为了确定最合适的检测方法，在14名健康志愿者的外周血单核细胞（PBMC）中，使用破伤风类毒素比较4种不同方法在回忆接种前后检测细胞因子反应的能力。选择ELISA、ELISPOT、胞内检测和实时RT-PCR来测量接种前后的IFN-γ产生。就记忆T细胞状态（接种前）的检测而言，我们发现ELISPOT是区分TT诱导反应和自发反应最敏感的方法。另一方面，胞内细胞因子检测是检测TT接种再刺激效应最有效的方法。
