PROTEIN-SYNTHESIS AND RELEASE BY CULTURED SCHISTOSOMULA OF SCHISTOSOMA-MANSONI
PROTEIN-SYNTHESIS AND RELEASE BY CULTURED SCHISTOSOMULA OF SCHISTOSOMA-MANSONI
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DOI：
10.1017/s0031182000079245
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发表时间：
1993-09-01
期刊：
影响因子：
2.4
通讯作者：
WILSON, RA
中科院分区：
文献类型：
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作者：
HARROP, R;WILSON, RA
曼氏血吸虫肺囊蚴是照射尾蚴单次免疫小鼠保护性免疫的靶点。由于效应子反应是T细胞介导的，它们的启动需要从完整的寄生虫释放抗原。我们已经使用了生物合成标记技术与[S-35]蛋氨酸，前和后的转化的尾蚴，分析蛋白质合成和释放的动力学的棘球蚴。此外，寄生虫的可溶性部分中存在的蛋白质和在体外培养过程中释放的蛋白质已被表征。在为期7天的培养期间，来自标记的尾蚴的童虫在转化后的第一个3小时内蛋白质损失最快。两种蛋白质的分子量为61和20 kDa的占主导地位，并可能对应于蛋白水解活性的领域。分析转化后标记的胚泡的蛋白质合成速率，揭示了四个不同的阶段，这可能与体内发生的发育过程有关。在第一个24小时，合成是非常低的，增加到一个平台，然后上升到一个高峰，在第8天，此后的速度迅速下降。虽然在寄生虫体的可溶性组分中检测到一些阶段特异性的蛋白质合成，但培养的幼虫释放的蛋白质的模式非常均匀。通过放射自显影法检测到至少15种蛋白质，其中61、45和20 kDa处的条带特别突出。这些蛋白质作为保护性免疫应答的潜在介质值得进一步研究。
The lung schistosomulum of Schistosoma mansoni is the target of protective immunity in mice singly vaccinated with irradiated cercariae. Since the effector responses are T cell-mediated, their initiation requires the release of antigens from the intact parasite. We have used the technique of biosynthetic labelling with [S-35]methionine, before and after transformation of the cercariae, to analyse the kinetics of protein synthesis and release by the schistosomulum. In addition, the proteins present in the soluble fraction of the parasite and those released during in vitro culture have been characterized. During a 7-day culture period schistosomula derived from labelled cercariae lost proteins most rapidly within the first 3 h after transformation. Two proteins of molecular weight 61 and 20 kDa were dominant and may correspond to areas of proteolytic activity. Analysis of the rate of protein synthesis of schistosomula labelled after transformation revealed four different phases, which may relate to the developmental processes occurring in vivo. During the first 24 h, synthesis was very low, increasing to a plateau and then rising to a peak at day 8; thereafter the rate declined rapidly. Whilst some stage-specific synthesis of proteins was detected in the soluble fractions of the parasite bodies, the pattern of proteins released by cultured larvae was remarkably uniform. At least 15 proteins were detected by autoradiography with bands at 61, 45 and 20 kDa being particularly prominent. These proteins merit further study as potential mediators of the protective immune response.