Nagel Angela, Möbs Christian, Raifer Hartmann, Wiendl Heinz, Hertl Michael, Eming Rüdiger
Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany; Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany.
PLoS One. 2014 Oct 16;9(10):e110138. doi: 10.1371/journal.pone.0110138. eCollection 2014.
The majority of clinical studies requires extensive management of human specimen including e.g. overnight shipping of blood samples in order to convey the samples in a central laboratory or to simultaneously analyze large numbers of patients. Storage of blood samples for periods of time before in vitro/ex vivo testing is known to influence the antigen expression on the surface of lymphocytes. In this context, the present results show for the first time that the T cell antigen CD3 can be substantially detected on the surface of human B cells after ex vivo storage and that the degree of this phenomenon critically depends on temperature and duration after blood withdrawal. The appearance of CD3 on the B cell surface seems to be a result of contact-dependent antigen exchange between T and B lymphocytes and is not attributed to endogenous production by B cells. Since cellular subsets are often classified by phenotypic analyses, our results indicate that ex vivo cellular classification in peripheral blood might result in misleading interpretations. Therefore, in order to obtain results reflecting the in vivo situation, it is suggested to minimize times of ex vivo blood storage after isolation of PBMC. Moreover, to enable reproducibility of results between different research groups and multicenter studies, we would emphasize the necessity to specify and standardize the storage conditions, which might be the basis of particular findings.
大多数临床研究需要对人体样本进行广泛管理，包括例如将血样隔夜运送，以便将样本送达中心实验室或同时分析大量患者。已知在体外/离体测试前将血样储存一段时间会影响淋巴细胞表面的抗原表达。在此背景下，本研究结果首次表明，离体储存后可在人B细胞表面大量检测到T细胞抗原CD3，且这种现象的程度严重依赖于采血后的温度和时长。B细胞表面CD3的出现似乎是T淋巴细胞与B淋巴细胞之间接触依赖性抗原交换的结果，而非B细胞内源性产生。由于细胞亚群通常通过表型分析进行分类，我们的结果表明外周血离体细胞分类可能会导致误导性的解读。因此，为了获得反映体内情况的结果，建议在分离外周血单个核细胞（PBMC）后尽量缩短血样离体储存时间。此外，为了确保不同研究团队和多中心研究结果的可重复性，我们强调必须明确并规范储存条件，这可能是特定研究结果的基础。
