Vaquero C, Sanceau J, Sondermeyer P, Falcoff R
Nucleic Acids Res. 1984 Mar 26;12(6):2629-40. doi: 10.1093/nar/12.6.2629.
Exposure of human lymphocytes to a mitogen induces the appearance of newly synthesized RNAs and proteins. This study describes the changes in overall synthesis as measured by pulse labelling of PHA treated lymphocytes as well as a qualitative analysis of the protein synthetic patterns "in vivo" and "in vitro". Both the levels of RNA and protein synthesis increase drastically in PHA stimulated cells, while cultures incubated without mitogen remained at background levels. The low translational activity in control cells is not due to the absence of messengers since the extracted RNAs clearly direct the synthesis of high molecular weight proteins when translated "in vitro". A number of qualitative differences are seen in the "in vitro" translation of RNA extracted from induced and non-induced lymphocytes, although the apparent protein synthetic pattern "in vivo" remains identical. The secretion of IFN- gamma is one of the newly expressed functions in stimulated lymphocytes and therefore has been studied more detailed in a time-course of the messenger level compared to the secreted activity of the medium. A specific probe was used to quantitate in Northern blot's the accumulation of mRNA coding for IFN- gamma.
将人淋巴细胞暴露于促有丝分裂原会诱导新合成的RNA和蛋白质出现。本研究描述了通过对PHA处理的淋巴细胞进行脉冲标记测量的总体合成变化，以及对“体内”和“体外”蛋白质合成模式的定性分析。在PHA刺激的细胞中，RNA和蛋白质合成水平均急剧增加，而未用促有丝分裂原培养的细胞则维持在背景水平。对照细胞中低翻译活性并非由于缺乏信使RNA，因为提取的RNA在“体外”翻译时能清楚地指导高分子量蛋白质的合成。从诱导和未诱导的淋巴细胞中提取的RNA在“体外”翻译时存在一些定性差异，尽管“体内”明显的蛋白质合成模式保持相同。γ干扰素的分泌是受刺激淋巴细胞中新表达的功能之一，因此与培养基的分泌活性相比，在信使水平的时间进程中对其进行了更详细的研究。使用特异性探针在Northern印迹中定量编码γ干扰素的mRNA的积累。
