Graziano M, St-Pierre Y, Beauchemin C, Desrosiers M, Potworowski E F
Centre de Recherche en Immunologie, Institut Armand-Frappier, Laval, Quebec, Canada.
Exp Cell Res. 1998 Apr 10;240(1):75-85. doi: 10.1006/excr.1997.3900.
The fate of thymic emigrants had so far been studied using a variety of markers, each of which had inherent limitations as to stability, toxicity, or selectivity. We describe a new technique which relies on the in vivo injection of CFSE, an esterified vital dye hitherto used at 80 times lower concentrations for in vitro cell labeling. We show that CFSE labels a representative sample of all thymocyte subsets and that these migrate at a rate of approximately 2-3 x 10(6) cells/day to peripheral lymphoid organs. We show that they enter lymph nodes at day 1 postinjection and stay for at least 21 days, whereas the turnover in the spleen is more rapid. We also show by immunohistochemistry, using peroxidase-labeled anti-FITC antibodies, that CFSE-labeled thymic emigrants are confined to T-dependent areas of peripheral lymphoid organs.
迄今为止，人们使用了多种标记物来研究胸腺迁出细胞的命运，每种标记物在稳定性、毒性或选择性方面都有其固有的局限性。我们描述了一种新技术，该技术依赖于体内注射CFSE，CFSE是一种酯化活性染料，迄今为止在体外细胞标记中使用的浓度要低80倍。我们发现CFSE标记了所有胸腺细胞亚群的代表性样本，并且这些细胞以大约2 - 3×10⁶个细胞/天的速度迁移到外周淋巴器官。我们发现它们在注射后第1天进入淋巴结，并停留至少21天，而在脾脏中的更新速度更快。我们还通过免疫组织化学方法，使用过氧化物酶标记的抗FITC抗体，表明CFSE标记的胸腺迁出细胞局限于外周淋巴器官的T细胞依赖区。
