细胞外囊泡(EV)是由几乎所有活着的王国产生的,在细胞间通讯过程中起着至关重要的作用。电动汽车对病原体尤其重要,作为恶性疟原虫(Pf)寄生虫,导致人类疟疾的主要物种。疟疾寄生虫能够通过在电动汽车内输送不同的货物成分,从远处调节宿主的免疫反应。如蛋白质和核酸。我们先前已经表明,成像流式细胞术(IFC)可以有效地用于监测宿主人单核细胞对疟疾衍生的EV的不同货物成分的摄取。这里,我们进一步采取这种方法,并证明我们可以通过监测其在免疫系统的两个不同的受体细胞内的分布来直接调查货物分配模式随时间的动态。单核细胞与巨噬细胞。通过染色囊泡的RNA货物并监测信号,我们能够评估其递送的动力学并测量内化后货物分布的不同参数。有趣的是,我们发现,虽然电动汽车的摄取水平相似,RNA货物分布的信号模式在这两种受体免疫细胞之间显著不同。我们的结果表明,该方法可用于研究吸收到不同类型细胞后囊泡货物的分布动力学。这对于我们理解病原体来源的囊泡与其宿主受体细胞之间的复杂界面中囊泡内化后货物成分的命运可以显着有益。
Extracellular vesicles (EVs) are produced by across almost all the living kingdoms and play a crucial role in cell-cell communication processes. EVs are especially important for pathogens, as Plasmodium falciparum (Pf) parasite, the leading causing species in human malaria. Malaria parasites are able to modulate the host immune response from a distance via delivering diverse cargo components inside the EVs, such as proteins and nucleic acids. We have previously shown that imaging flow cytometry (IFC) can be effectively used to monitor the uptake of different cargo components of malaria derived EVs by host human monocytes. Here, we take this approach one step further and demonstrate that we can directly investigate the dynamics of the cargo distribution pattern over time by monitoring its distribution within two different recipient cells of the immune system, monocytes vs macrophages. By staining the RNA cargo of the vesicles and monitor the signal we were able to evaluate the kinetics of its delivery and measure different parameters of the cargo\'s distribution post internalization. Interestingly, we found that while the level of the EV uptake is similar, the pattern of the signal for RNA cargo distribution is significantly different between these two recipient immune cells. Our results demonstrate that this method can be applied to study the distribution dynamics of the vesicle cargo post uptake to different types of cells. This can benefit significantly to our understanding of the fate of cargo components post vesicle internalization in the complex interface between pathogen-derived vesicles and their host recipient cells.