Westermann J, Puskas Z, Pabst R
Centre of Anatomy, Medical School of Hanover, FRG.
Scand J Immunol. 1988 Aug;28(2):203-10. doi: 10.1111/j.1365-3083.1988.tb02432.x.
Thoracic duct lymphocytes (TDL) were labelled with fluorescein isothiocyanate (FITC) and injected into normal, unanaesthetized rats with either a central venous catheter or a thoracic duct cannula. The blood transit time and the appearance in the lymph was calculated and then the percentages of B, T, T helper, and T suppressor lymphocytes were determined with monoclonal antibodies (Ox12, Ox19, W3/25, and Ox8, respectively). The blood transit time of all subsets was about 30 min. However, the percentage of B lymphocytes from 5 min after injection onwards is reduced (14.5 +/- 1.9%) compared to the injected TDL (32.7 +/- 1.7%). These cells are not in the lung vascular pool. The recovery of FITC-labelled TDL in the thoracic duct within 48 h is much higher (56.8 +/- 5.3% of the injected lymphocytes) than in previous studies using radioactive markers. B lymphocytes appear later and in a reduced number in the thoracic duct. The mean transit time is 26 h and the recovery 31.5 +/- 4.2% in contrast to T lymphocytes (18 h and 66.3 +/- 6.5%, respectively). The technique of combining FITC in vitro labelling with surface staining after completion of migration does not interfere with lymphocyte migration. It can therefore be used to study the migration of lymphocyte subsets in normal, untreated animals.
用异硫氰酸荧光素（FITC）标记胸导管淋巴细胞（TDL），并通过中心静脉导管或胸导管插管将其注入正常、未麻醉的大鼠体内。计算血液通过时间和在淋巴中的出现情况，然后分别用单克隆抗体（分别为Ox12、Ox19、W3/25和Ox8）测定B淋巴细胞、T淋巴细胞、辅助性T淋巴细胞和抑制性T淋巴细胞的百分比。所有亚群的血液通过时间约为30分钟。然而，与注入的TDL（32.7±1.7%）相比，注射后5分钟起B淋巴细胞的百分比降低（14.5±1.9%）。这些细胞不在肺血管池中。48小时内胸导管中FITC标记的TDL的回收率（占注入淋巴细胞的56.8±5.3%）比以前使用放射性标记的研究高得多。B淋巴细胞在胸导管中出现较晚且数量减少。平均通过时间为26小时，回收率为31.5±4.2%，而T淋巴细胞分别为18小时和66.3±6.5%。将体外FITC标记与迁移完成后的表面染色相结合的技术不会干扰淋巴细胞迁移。因此，它可用于研究正常未处理动物中淋巴细胞亚群的迁移。
