Ding M, Robinson J M, Behrens B C, Vandré D D
Department of Cell Biology, Neurobiology, and Anatomy, Ohio State University, Columbus 43210-1239, USA.
Eur J Cell Biol. 1995 Mar;66(3):234-45.
The microtubule cytoskeleton of human leukocytes has been difficult to study, in part, due to the lack of a reliable protocol for the indirect immunofluorescence staining of microtubules in these cells. We report here the development of a simple and reliable immunocytochemical labeling protocol for the examination of microtubules in leukocytes including monocytes, neutrophils, and eosinophils. The dynamic properties of microtubules in both monocytes and neutrophils were examined by indirect immunofluorescence staining of cells following exposure to nocodazole. Nocodazole-induced depolymerization is extremely rapid in both cell types, as is the regrowth of microtubules following removal of the nocodazole. Rapid reorganization of the microtubule cytoskeleton was also observed in neutrophils undergoing chemotactic stimulation. Bundling of microtubules was observed in both monocytes and neutrophils isolated from patients undergoing taxol infusion chemotherapy. The taxol-induced bundles were transient in nature as they were absent from samples collected 48 h following the completion of the taxol infusion. These results demonstrate the unique dynamic properties of leukocyte microtubules and indicate that they can be altered in vivo. The development of this staining protocol should allow for the further analysis of leukocyte microtubules as related to the normal functional response of these cells and form the basis for correlating alterations in microtubule dynamics with the effects of taxol on leukocyte function.
人类白细胞的微管细胞骨架一直难以研究，部分原因是缺乏一种可靠的方案来对这些细胞中的微管进行间接免疫荧光染色。我们在此报告一种简单可靠的免疫细胞化学标记方案的开发，用于检查包括单核细胞、中性粒细胞和嗜酸性粒细胞在内的白细胞中的微管。通过对暴露于诺考达唑后的细胞进行间接免疫荧光染色，研究了单核细胞和中性粒细胞中微管的动态特性。在这两种细胞类型中，诺考达唑诱导的解聚极其迅速，去除诺考达唑后微管的重新生长也是如此。在经历趋化刺激的中性粒细胞中也观察到微管细胞骨架的快速重组。在接受紫杉醇输注化疗的患者分离出的单核细胞和中性粒细胞中均观察到微管的成束现象。紫杉醇诱导的微管束本质上是短暂的，因为在紫杉醇输注完成后48小时采集的样本中不存在这种微管束。这些结果证明了白细胞微管独特的动态特性，并表明它们在体内可以被改变。这种染色方案的开发应该有助于进一步分析与这些细胞正常功能反应相关的白细胞微管，并为将微管动力学的改变与紫杉醇对白细胞功能的影响相关联奠定基础。
