Ilvonen Petra, Pusa Reetta, Härkönen Kai, Laitinen Saara, Impola Ulla
Finnish Red Cross Blood Service Helsinki Finland.
J Extracell Biol. 2024 Jan 2;3(1):e130. doi: 10.1002/jex2.130. eCollection 2024 Jan.
Blood-derived extracellular vesicles (EVs) hold great therapeutic potential. As blood contains mixed EV populations, it is challenging to study EVs originating from different cells separately. Blood cell concentrates manufactured in blood banks offer an excellent non-invasive source of blood cell-specific EV populations. To study blood cell-specific EVs, we isolated EVs from platelet (TREVs) and red blood cell (EryEVs) concentrates and characterized them using nanoparticle tracking analysis, imaging flow cytometry, electron microscopy and western blot analysis and co-cultured them with peripheral blood mononuclear cells (PBMCs). Our aim was to use imaging flow cytometry to investigate EV interaction with PBMCs as well as study their effects on T-lymphocyte populations to better understand their possible biological functions. As a conclusion, TREVs interacted with PBMCs more than EryEVs. Distinctively, TREVs were uptaken into CD11c+ monocytes rapidly and into CD19+ B-lymphocytes in 24 h. EryEVs were not uptaken into CD11c+ monocytes before the 24-h time point, and they were only seen on the surface of lymphocytes. Neither TREVs nor EryEV were uptaken into CD3+ T-lymphocytes and no effect on T-cell populations was detected. We have previously seen similar differences in targeting PC-3 cancer cells. Further studies are needed to address the functional properties of blood cell concentrate-derived EVs. This study demonstrates that imaging flow cytometry can be used to study the distinctive differences in the interaction and uptake of EVs. Considering our current and previous results, EVs present a new valuable component for the future development of blood-derived therapeutics.
血液来源的细胞外囊泡（EVs）具有巨大的治疗潜力。由于血液中含有混合的EV群体，单独研究源自不同细胞的EV具有挑战性。血库中制备的血细胞浓缩物提供了血细胞特异性EV群体的优质非侵入性来源。为了研究血细胞特异性EV，我们从血小板浓缩物（TREVs）和红细胞浓缩物（EryEVs）中分离出EV，并使用纳米颗粒跟踪分析、成像流式细胞术、电子显微镜和蛋白质免疫印迹分析对其进行表征，然后将它们与外周血单核细胞（PBMCs）共培养。我们的目的是使用成像流式细胞术研究EV与PBMCs的相互作用，并研究它们对T淋巴细胞群体的影响，以更好地了解它们可能的生物学功能。结论是，TREVs与PBMCs的相互作用比EryEVs更多。值得注意的是，TREVs迅速被CD11c+单核细胞摄取，并在24小时内被CD19+B淋巴细胞摄取。在24小时时间点之前，EryEVs未被CD11c+单核细胞摄取，仅在淋巴细胞表面可见。TREVs和EryEVs均未被CD3+T淋巴细胞摄取，且未检测到对T细胞群体的影响。我们之前在靶向PC-3癌细胞方面也看到了类似的差异。需要进一步研究来探讨血细胞浓缩物来源的EV的功能特性。这项研究表明，成像流式细胞术可用于研究EV相互作用和摄取的显著差异。综合我们目前和之前的结果，EVs为血液衍生疗法的未来发展提供了一种新的有价值的成分。
