Zmievskaya Ekaterina A, Mukhametshin Sabir A, Ganeeva Irina A, Gilyazova Elvina M, Siraeva Elvira T, Kutyreva Marianna P, Khannanov Artur A, Yuan Youyong, Bulatov Emil R
Institute of Fundamental Medicine and Biology, Kazan Federal University, 420008 Kazan, Russia.
A.M. Butlerov Institute of Chemistry, Kazan Federal University, 420008 Kazan, Russia.
Biomedicines. 2024 Apr 20;12(4):919. doi: 10.3390/biomedicines12040919.
Cell therapy is at the forefront of biomedicine in oncology and regenerative medicine. However, there are still significant challenges to their wider clinical application such as limited efficacy, side effects, and logistical difficulties. One of the potential approaches that could overcome these problems is based on extracellular vesicles (EVs) as a cell-free therapy modality. One of the major obstacles in the translation of EVs into practice is their low yield of production, which is insufficient to achieve therapeutic amounts. Here, we evaluated two primary approaches of artificial vesicle induction in primary T cells and the SupT1 cell line-cytochalasin B as a chemical inducer and ultrasonication as a physical inducer. We found that both methods are capable of producing artificial vesicles, but cytochalasin B induction leads to vesicle yield compared to natural secretion, while ultrasonication leads to a three-fold increase in particle yield. Cytochalasin B induces the formation of vesicles full of cytoplasmic compartments without nuclear fraction, while ultrasonication induces the formation of particles rich in membranes and membrane-related components such as CD3 or HLAII proteins. The most effective approach for T-cell induction in terms of the number of vesicles seems to be the combination of anti-CD3/CD28 antibody activation with ultrasonication, which leads to a seven-fold yield increase in particles with a high content of functionally important proteins (CD3, granzyme B, and HLA II).
细胞疗法在肿瘤学和再生医学的生物医学领域处于前沿地位。然而，其更广泛的临床应用仍面临重大挑战，如疗效有限、副作用和后勤困难等。一种可能克服这些问题的潜在方法是基于细胞外囊泡（EVs）作为一种无细胞治疗方式。将EVs转化为实际应用的主要障碍之一是其产量低，不足以达到治疗剂量。在此，我们评估了在原代T细胞和SupT1细胞系中诱导人工囊泡的两种主要方法——细胞松弛素B作为化学诱导剂和超声处理作为物理诱导剂。我们发现这两种方法都能够产生人工囊泡，但与自然分泌相比，细胞松弛素B诱导导致囊泡产量增加，而超声处理导致颗粒产量增加三倍。细胞松弛素B诱导形成充满细胞质成分而无核成分的囊泡，而超声处理诱导形成富含膜和膜相关成分（如CD3或HLAII蛋白）的颗粒。就囊泡数量而言，T细胞诱导的最有效方法似乎是抗CD3/CD28抗体激活与超声处理相结合，这导致富含功能重要蛋白质（CD3、颗粒酶B和HLA II）的颗粒产量增加七倍。
