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首页> 外文期刊>Cellular immunology >Induced cytokine response of human PMBC-cultures: correlation of gene expression and secretion profiling and the effect of cryopreservation.
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Induced cytokine response of human PMBC-cultures: correlation of gene expression and secretion profiling and the effect of cryopreservation.

机译：诱导人类PMBC培养的细胞因子反应：基因表达和分泌概况以及冷冻保存作用的相关性。

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摘要

The immune system is regulated by the complex interaction of multiple cytokines, which are secreted signaling molecules affecting other cells. In this work, we studied the cytokine response to several well-known stimulants, such as OKT-3, Con A, PWM, and SEB. Healthy donor cells (PBMCs) were cultivated for up to 72 h and the mRNA levels and cytokine release of four key cytokines (IL-2, IL-4, IFN-γ, and TNF-α) were analyzed by RT-PCR and bead-based multiplex analyses. The generated cytokine profiles showed characteristic expression patterns and secretion kinetics for each cytokine and substance. PWM/SEB and OKT-3 led to a very fast and long-lasting immune response, whereas Con A induced the slowest cytokine production. Cytokine concentrations also differed greatly. The highest IFN-γ concentration was 1000 times higher than the respective IL-4 concentration. Gene expression and cytokine concentration profiles were strongly correlated during the time course. The chronological response of the donors' cytokine profiles coincided, but showed individual characteristics regarding the strength of the cytokine release. The comparison of stimulation experiments using freshly isolated and cryopreserved PBMCs showed that, for the observation of an immunological response at early points in time, gene expression experiments are more reliable than the measurement of cytokines in the cell culture supernatant. However, the freezing of cells influences the response significantly. The measurement of secreted proteins is the superior method at later points in time.
机译：免疫系统受到多种细胞因子复杂相互作用的调节，这些因子是分泌的影响其他细胞的信号分子。在这项工作中，我们研究了细胞因子对几种众所周知的兴奋剂的反应，例如OKT-3，Con A，PWM和SEB。培养健康的供体细胞（PBMC）长达72小时，并通过RT-PCR和磁珠分析四种关键细胞因子（IL-2，IL-4，IFN-γ和TNF-α）的mRNA水平和细胞因子释放基于多元的分析。产生的细胞因子概况显示每种细胞因子和物质的特征性表达模式和分泌动力学。 PWM / SEB和OKT-3导致非常快速和持久的免疫反应，而Con A诱导最慢的细胞因子产生。细胞因子的浓度也相差很大。最高的IFN-γ浓度比相应的IL-4浓度高1000倍。在时间过程中，基因表达和细胞因子浓度分布密切相关。供体细胞因子分布的时间响应一致，但显示出有关细胞因子释放强度的个体特征。使用新鲜分离的和冷冻保存的PBMC进行刺激实验的比较表明，为了在早期观察免疫反应，基因表达实验比测量细胞培养上清液中的细胞因子更可靠。但是，细胞的冻结会显着影响反应。在以后的时间点，分泌蛋白的测量是一种更好的方法。

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