骨髓瘤来源的微颗粒对人外周血同种免疫单核细胞体外增殖和活力的影响

S. Milani, F. Yari
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摘要

背景:微颗粒(MPs)是从细胞膜释放出来的小囊泡。因此,这些含有活性分子来介导生物过程,包括细胞增殖和细胞周期进程。骨髓瘤细胞系与免疫B淋巴细胞的融合是杂交技术的关键步骤。MPs调节淋巴细胞增殖,从而促进B细胞扩增和成功永生化。人类同种免疫B细胞被认为是单克隆抗体的重要来源,外周血是B淋巴细胞的储存库。本研究旨在探讨骨髓瘤源性MPs在同种免疫外周血单个核细胞(PBMCs)增殖中的作用。材料和方法:在本实验研究中,我们采用超离心方法从三种不同的骨髓瘤细胞系U266、P3x63Ag8和SP2/0中分离MPs。从暴露于MPs的地中海贫血同种免疫患者的全血中提取PBMCs。然后用倒置显微镜和台盼蓝染色法检测细胞增殖和细胞活力。结果:来源于P3X63Ag8骨髓瘤细胞系的MPs对细胞增殖和活力的影响最大(p=0.0005)。SP2/0细胞系的MPs最初增加了pbmc的增殖,但在接下来的几周内活性细胞急剧减少。同样,U266细胞来源的MPs增加了PBMCs的增殖(p=0.0001),但这是接受P3x63Ag8来源MPs的组的一半。然而,处理过的细胞的活力在5周内几乎保持不变。结论:综上所述,P3X63Ag8和U266衍生的MPs可提高PBMCs的生存能力。如果补充检查得到证实,这些MPs也可以作为B淋巴细胞增殖的潜在因子,从而有助于永生化和抗体的产生。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Effect of Myeloma-Derived Microparticles on in Vitro Proliferation and Viability of Alloimmune Human Peripheral Blood Mononuclear Cells
Background: Microparticles (MPs) are small vesicles released from the cell membrane. Accordingly, these contain active molecules to mediate biological processes, including cell proliferation and cell cycle progression. The fusion of myeloma cell lines with immunized B lymphocytes is a critical step of hybridoma technology. MPs modulate lymphoproliferation, thereby facilitating B cell expansion and successful immortalization. Human alloimmune B cells are considered valuable sources of monoclonal antibodies, and peripheral blood is a pool of B lymphocytes. The present study aimed to investigate the role of myeloma-derived MPs in the proliferation of alloimmune peripheral blood mononuclear cells (PBMCs). Materilas and Methods: In the current experimental study, ultracentrifugation isolated MPs from three different myeloma cell lines, including U266, P3x63Ag8, and SP2/0. PBMCs were extracted from the whole blood of a patient with thalassemia alloimmune exposed to MPs. Thereafter, both the proliferation and cell viability were evaluated using inverted microscopy and the trypan blue staining method. Results: MPs derived from the P3X63Ag8 myeloma cell line were shown to have the most effects on cell proliferation and viability (p=0.0005). MPs of the SP2/0 cell line initially increased the proliferation of PBMCs, but viable cells were drastically reduced in the following weeks. As well, U266 cell-derived MPs increased the proliferation of PBMCs (p=0.0001), but it was half of the group receiving P3x63Ag8 derived MPs. However, the viability of treated cells remained almost constant for 5-weeks. Conclusion: Altogether, the obtained data indicated that P3X63Ag8 and U266 derived MPs could increase the viability of PBMCs. If the complimentary examination is confirmed, these MPs could also be used as the potential agents in B lymphocyte proliferation, thereby helping in immortalization and antibody production.
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