Pfeifer R W, Irons R D
Int J Immunopharmacol. 1983;5(5):463-70. doi: 10.1016/0192-0561(83)90023-1.
The cytoskeleton plays a pivotal role in lectin-induced lymphocyte blastogenesis. Microtubule disrupting agents, many of which are sulfhydryl (SH) reagents, interfere with cytoskeletal-dependent cell functions including lymphocyte blastogenesis and agglutination. For example, hydroquinone (HQ) and N-ethylmaleimide (NEM) inhibit lectin-stimulated lymphocyte blastogenesis and agglutination at concentrations (10(-5)M) that do not reduce cell viability or ATP production. Indicative of the SH-specificity of these effects, only L-cysteine protects against HQ or NEM inhibition of blastogenesis and agglutination. Other compounds, including L-serine, DL-lysine and imidazole, have no protective effect. These and other findings previously reported suggest a selective interaction of HQ, or its oxidation product, p-benzoquinone (p-BQ) with SH groups critical to early G1 events associated with lymphocyte activation. These compounds show similar SH specificity in inhibiting microtubule assembly in vitro. The subcellular target specificity (cytoskeleton) exhibited by these compounds was compared to that of Adriamycin (ADR), a complex polycyclic quinone with known immunotoxic activity. ADR inhibited microtubule assembly in vitro and inhibited lymphocyte blastogenesis, however, these effects were not correlated with a loss of agglutination nor was toxicity protected against by the addition of SH compounds. The combination of cell culture methods together with application of techniques to measure microtubule assembly in vitro provides an effective means to discriminate between agents that selectively interfere with cytoskeletal-dependent function and those producing non-specific effects associated with cell death, such as decreased energy production or increased membrane permeability.
细胞骨架在凝集素诱导的淋巴细胞增殖中起关键作用。许多破坏微管的试剂（其中许多是巯基（SH）试剂）会干扰包括淋巴细胞增殖和凝集在内的细胞骨架依赖性细胞功能。例如，对苯二酚（HQ）和N-乙基马来酰亚胺（NEM）在不降低细胞活力或ATP生成的浓度（10⁻⁵M）下抑制凝集素刺激的淋巴细胞增殖和凝集。这些效应具有SH特异性的一个标志是，只有L-半胱氨酸能防止HQ或NEM对增殖和凝集的抑制。其他化合物，包括L-丝氨酸、DL-赖氨酸和咪唑，没有保护作用。先前报道的这些及其他发现表明，HQ或其氧化产物对苯醌（p-BQ）与对淋巴细胞激活相关的早期G1事件至关重要的SH基团存在选择性相互作用。这些化合物在体外抑制微管组装时表现出类似的SH特异性。将这些化合物表现出的亚细胞靶点特异性（细胞骨架）与阿霉素（ADR）进行了比较，阿霉素是一种具有已知免疫毒性活性的复杂多环醌。ADR在体外抑制微管组装并抑制淋巴细胞增殖，然而，这些效应与凝集的丧失无关，添加SH化合物也不能防止其毒性。细胞培养方法与体外测量微管组装技术相结合，为区分选择性干扰细胞骨架依赖性功能的试剂和产生与细胞死亡相关的非特异性效应（如能量生成减少或膜通透性增加）的试剂提供了一种有效方法。
