Eisenthal A, Marder O, Lifschitz-Mercer B, Skornick Y, Tirosh R, Weinreb A, Deutsch M
Pathology Institute, Sourasky Medical Center, Tel-Aviv, Israel.
Cell Struct Funct. 1996 Jun;21(3):159-66. doi: 10.1247/csf.21.159.
We have previously reported that the exposure of human peripheral blood lymphocytes (PBL) to a variety of stimulants caused rapid changes in intracellular fluorescein fluorescence polarization (IFFP) in the activated cells. In the present study we further analyzed possible mechanisms responsible for the changes in IFFP in PBL exposed to phytohaemagglutinin (PHA) and anti-CD3 antibody. By employing several agents which are known to affect the polymerization of the cytoskeleton we showed that both cytochalasin B, which regulates the microfilaments structure, and vinblastine and colchicine, which affect the microtubules, completely abolished the changes induced in IFFP of human PBL by both PHA and anti-CD3. This effect was dose dependent and was noted at concentrations ranging from 10 to 100 microM of cytochalasin B and 10 microM of vinblastine and colchicine. The effect of these cytoskeleton modulators occurred within 20 minutes after the initiation of activation with PHA. Our results indicate that activation with PHA and anti-CD3 causes early changes in the microtubules and microfilaments components of the cytoskeleton. The possible application of IFFP measurement in analyzing early changes in the cytoskeleton following cell activation is discussed.
我们之前曾报道，人类外周血淋巴细胞（PBL）暴露于多种刺激物会导致活化细胞内的荧光素荧光偏振（IFFP）迅速变化。在本研究中，我们进一步分析了暴露于植物血凝素（PHA）和抗CD3抗体的PBL中IFFP变化的可能机制。通过使用几种已知会影响细胞骨架聚合的药物，我们发现，调节微丝结构的细胞松弛素B以及影响微管的长春花碱和秋水仙碱，都完全消除了PHA和抗CD3诱导的人类PBL的IFFP变化。这种效应呈剂量依赖性，在细胞松弛素B浓度为10至100微摩尔以及长春花碱和秋水仙碱浓度为10微摩尔时即可观察到。这些细胞骨架调节剂的作用在PHA激活开始后20分钟内出现。我们的结果表明，PHA和抗CD3激活会导致细胞骨架的微管和微丝成分发生早期变化。本文还讨论了IFFP测量在分析细胞激活后细胞骨架早期变化中的可能应用。
