Laboratorio de Inmunología y estrés de Organismos Acuáticos, Instituto de Patología Animal, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Valdivia, Chile.
Laboratorio de Fisiología de peces, Instituto de Ciencias Marinas y Limnológicas, Facultad de Ciencias, Universidad Austral de Chile, Valdivia, Chile.
Front Immunol. 2022 Apr 14;13:849752. doi: 10.3389/fimmu.2022.849752. eCollection 2022.
The innate immune system can limit the growth of invading pathogens by depleting micronutrients at a cellular and tissue level. However, it is not known whether nutrient depletion mechanisms discriminate between living pathogens (which require nutrients) and pathogen-associated molecular patterns (PAMPs) (which do not). We stimulated SHK-1 cells with different PAMPs (outer membrane vesicles of "OMVs", protein extract of "TP" and lipopolysaccharides of "LPS") isolated from and evaluated transcriptional changes in nutritional immunity associated genes. Our experimental treatments were: Control (SHK-1 stimulated with bacterial culture medium), OMVs (SHK-1 stimulated with 1μg of outer membrane vesicles), TP (SHK-1 stimulated with 1μg of total protein extract) and LPS (SHK-1 stimulated with 1μg of lipopolysaccharides). Cells were sampled at 15-, 30-, 60- and 120-minutes post-stimulation. We detected increased transcription of , , , and in all three experimental conditions and increased transcription of in cells stimulated with OMVs and TP, but not LPS. Additionally, we observed generally increased transcription of , , , and in all three experimental conditions, but we also detected decreased transcription of these markers in cells stimulated with TP and LPS at specific time points. Our results demonstrate that SHK-1 cells stimulated with PAMPs increase transcription of markers involved in the transport, uptake, storage and regulation of micronutrients such as iron, manganese and zinc.
固有免疫系统可以通过消耗细胞和组织水平的微量营养素来限制入侵病原体的生长。然而，目前尚不清楚营养物质耗竭机制是否区分活病原体（需要营养物质）和病原体相关分子模式（PAMPs）（不需要）。我们用不同的 PAMPs（“OMVs”的外膜囊泡、“TP”的蛋白质提取物和“LPS”的脂多糖）刺激 SHK-1 细胞，并评估与营养免疫相关基因的转录变化。我们的实验处理为：对照（用细菌培养基刺激的 SHK-1）、OMVs（用 1μg 外膜囊泡刺激的 SHK-1）、TP（用 1μg 总蛋白提取物刺激的 SHK-1）和 LPS（用 1μg 脂多糖刺激的 SHK-1）。在刺激后 15、30、60 和 120 分钟采集细胞样本。我们在所有三种实验条件下都检测到 、 、 、 和 的转录增加，在 OMVs 和 TP 刺激的细胞中检测到 的转录增加，但 LPS 刺激的细胞中没有。此外，我们观察到所有三种实验条件下普遍增加了 、 、 、 和 的转录，但也检测到在特定时间点用 TP 和 LPS 刺激的细胞中这些标志物的转录减少。我们的结果表明，用 PAMPs 刺激的 SHK-1 细胞增加了参与铁、锰和锌等微量营养素的运输、摄取、储存和调节的标志物的转录。
