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首页> 外文期刊>Biotechnology Progress >Hyperosmotic stress in murine Hybridoma cells: Effects on antibody transcription, translation, posttranslational processing, and the cell cycle
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Hyperosmotic stress in murine Hybridoma cells: Effects on antibody transcription, translation, posttranslational processing, and the cell cycle

机译：鼠杂交瘤细胞中的高渗应激：对抗体转录，翻译，翻译后加工和细胞周期的影响

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Mechanisms for increased antibody production in batch cultures of murine hybridoma cells in response to hyperosmotic stress were investigated. The rates of immunoglobulin transcription and protein translation and posttranslational processing were determined in control and hyperosmotic cultures. Changes in immunoglobulin transcription played a minor role in the increase in antibody production in response to hyperosmotic stress. In contrast, protein translation increased substantially in response to osmotic stress. However, the antibody translation rate remained relatively constant after correcting for the overall increase in protein translation. Cell size and intracellular antibody pool also increased in response to hyperosmolarity. The intracellular antibody pool increased proportionately with the increase in cell size, indicating that hyperosmotic cultures do not selectively increase their intracellular antibody population. Changes in cell cycle distribution in response to osmotic stress and the relationship between the cell cycle and antibody production were also evaluated. Hyperosmotic stress altered the cell cycle distribution, increasing the fraction of the cells in S-phase. However, this change was uncorrelated with the increase in antibody production rate. Immunoglobulin degradation was relatively low (similar to15%) and remained largely unchanged in response to hyperosmotic stress. There was no apparent increase in immunoglobulin stability as a result of osmotic stress. Antibody secretion rates increased approximately 50% in response to osmotic stress, with a commensurate increase in the antibody assembly rate. The rate of transit through the entire posttranslational processing apparatus increased, particularly for immunoglobulin light chains. The levels of endoplasmic reticulum chaperones did not increase as a fraction of the total cellular protein but were increased on a per cell basis as the result of an increase in total cellular protein. A difference in the interactions between the immunoglobulin heavy chains and BiP/GRP78 was observed in response to hyperosmotic conditions. This change in interaction may be correlated with the decrease in transit time through the posttranslational pathways. The increase in the posttranslational processing rate appears to be commensurate with the increase in antibody production in response to hyperosmotic stress.
机译：研究了在高渗胁迫下鼠杂交瘤细胞分批培养中增加抗体产生的机制。在对照和高渗培养物中确定免疫球蛋白转录，蛋白质翻译和翻译后加工的速率。免疫球蛋白转录的变化在响应高渗应激的抗体产生增加中起次要作用。相反，响应于渗透压，蛋白质翻译显着增加。然而，在校正蛋白质翻译的总体增加之后，抗体翻译速率保持相对恒定。细胞的大小和细胞内抗体库也响应高渗性而增加。细胞内抗体库随细胞大小的增加成比例地增加，表明高渗培养物不会选择性地增加其细胞内抗体的数量。还评估了响应渗透压的细胞周期分布的变化以及细胞周期与抗体产生之间的关系。高渗胁迫改变了细胞周期分布，增加了S期细胞的比例。但是，这种变化与抗体产生速率的增加无关。免疫球蛋白的降解相对较低（约15％），并且在高渗胁迫下仍保持基本不变。由于渗透胁迫，免疫球蛋白的稳定性没有明显增加。响应于渗透压力，抗体分泌率提高了约50％，抗体装配率也相应提高。通过整个翻译后加工设备的转运速率增加，特别是对于免疫球蛋白轻链而言。内质网伴侣分子的水平没有作为总细胞蛋白的一部分而增加，而是由于总细胞蛋白的增加而在每个细胞的基础上增加。响应高渗条件，观察到免疫球蛋白重链与BiP / GRP78之间相互作用的差异。相互作用的这种改变可能与通过翻译后途径的转运时间减少有关。翻译后加工速率的增加似乎与响应高渗应激的抗体产生的增加相对应。

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