Salameire D, Le Bris Y, Fabre B, Fauconnier J, Solly F, Pernollet M, Bonnefoix T, Leroux D, Plumas J, Jacob M C
Université Joseph Fourier Grenoble 1, F-38000, France.
Cytometry A. 2009 Sep;75(9):743-51. doi: 10.1002/cyto.a.20767.
Analysis of the T-cell receptor (TCR) repertoire by flow cytometry proved to be relevant for investigating T-cell diversity and detecting reactive cells in blood samples. We used this approach to characterize non-malignant T-lymphocytes in lymph nodes and give insights into their origin. The TCR repertoire of CD4+ and CD8+ T-cells from 81 lymph nodes was analyzed with a four-color flow cytometer using a wide panel of 25 anti-Vbeta monoclonal antibodies. Flow cytometry proved to be a useful and informative technique. We demonstrated a diversified TCR-Vbeta repertoire, and only low level expansions, in 53% of the samples. They involved nearly all Vbeta families, were more frequent in the CD8+ subset of older patients, but were not related to pathology. No evidence could be demonstrated in favor of stimulation by common antigens. Interestingly, the TCR-Vbeta repertoire proved to be very similar in lymph nodes and blood samples. Our results argue that in the cases studied, lymph node enlargement is mainly due to an increased homing of circulating T-cells. They also provide reference values for expression of 25 TCR-Vbeta in lymph nodes, which could serve as a basis for further applications in diagnosis of T-cell lymphoproliferative disorders.
通过流式细胞术分析T细胞受体（TCR）库被证明与研究T细胞多样性以及检测血液样本中的反应性细胞相关。我们采用这种方法来表征淋巴结中的非恶性T淋巴细胞，并深入了解其来源。使用25种抗Vβ单克隆抗体的广泛面板，通过四色流式细胞仪分析了来自81个淋巴结的CD4 +和CD8 + T细胞的TCR库。流式细胞术被证明是一种有用且信息丰富的技术。我们在53%的样本中证明了多样化的TCR-Vβ库，且只有低水平的扩增。它们涉及几乎所有的Vβ家族，在老年患者的CD8 +亚群中更常见，但与病理学无关。没有证据表明存在共同抗原刺激。有趣的是，TCR-Vβ库在淋巴结和血液样本中被证明非常相似。我们的结果表明，在所研究的病例中，淋巴结肿大主要是由于循环T细胞归巢增加所致。它们还提供了25种TCR-Vβ在淋巴结中表达的参考值，这可为T细胞淋巴增殖性疾病诊断的进一步应用提供基础。
