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首页> 美国卫生研究院文献>British Journal of Cancer >The effect of ricin B chain on the intracellular trafficking of an A chain immunotoxin.
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The effect of ricin B chain on the intracellular trafficking of an A chain immunotoxin.

机译：蓖麻毒素B链对A链免疫毒素的细胞内运输的影响。

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摘要

Covalent linkage of the A chain of ricin to the LICR-LOND-Fib75 monoclonal antibody produced an immunotoxin, Fib75-SS-ricin A, which demonstrated immunospecific toxicity to human bladder carcinoma cells in tissue culture (Forrester et al., 1984). The present studies have shown that ricin B chain potentiates the toxicity of the immunotoxin by two orders of magnitude and also significantly increases the rate of protein synthesis inhibition. Using immunoelectron microscopy, the receptor-mediated endocytosis and intracellular routing of the immunotoxin was studied with and without ricin B chain treatment after immunolocalisation of the conjugate. Fib75-SS-ricin A was internalised by the EJ cells predominantly in uncoated pits and vesicles and directed to the endosomes. Some degradation of the complex appeared to take place in multivesicular endosomes at early timepoints and 24 h after internalisation, most of the immunotoxin was found in lysosomes. Some ricin A chain epitopes were detected in Golgi vesicles. Cells treated with immunotoxin and ricin B chain endocytosed the complex predominantly in coated pits and coated vesicles. Using pre-embedding immunoperoxidase techniques, ricin chains were found in the whole Golgi complex and most of the conjugate escaped lysosomal degradation. Internalised immunotoxin was recycled back to the plasma membrane in an active form associated with vesicles which appeared to be derived predominantly from multivesicular endosomes. A similar mode of recycling has recently been reported (McIntosh et al., 1990) for ricin holotoxin in the same cell line. These observations may explain the potentiating effect of toxin B chains in the antibody-directed targeting of toxin A chains.
机译：蓖麻毒蛋白的A链与LICR-LOND-Fib75单克隆抗体的共价连接产生了一种免疫毒素，即Fib75-SS-蓖麻毒蛋白A，它在组织培养中显示出对人膀胱癌细胞的免疫特异性毒性（Forrester等，1984）。目前的研究表明，蓖麻毒蛋白B链将免疫毒素的毒性增强了两个数量级，并且还显着提高了蛋白质合成抑制率。使用免疫电子显微镜，研究了结合物免疫定位后有无蓖麻毒素B链处理的受体介导的内吞作用和免疫毒素的细胞内途径。 Fib75-SS-蓖麻毒蛋白A主要由未包被的凹坑和囊泡中的EJ细胞内在化，并指向内体。复合物的某些降解似乎在早期的时间点和内化后24小时内在多囊泡内体中发生，大部分免疫毒素在溶酶体中发现。在高尔基囊泡中检测到一些蓖麻毒蛋白A链表位。用免疫毒素和蓖麻毒素B链处理的细胞主要在包被的凹坑和包被的囊泡中内吞复合物。使用包埋前免疫过氧化物酶技术，在整个高尔基体中发现了蓖麻毒蛋白链，大多数结合物都逃脱了溶酶体降解。内在的免疫毒素以与囊泡有关的活性形式再循环回质膜，囊泡似乎主要来自多囊泡内体。最近已经报道了在同一细胞系中蓖麻毒素全毒素的类似回收方式（McIntosh等，1990）。这些观察结果可以解释毒素B链在抗体定向的毒素A链靶向中的增强作用。

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