Wang Jiemin, Moosavizadeh Seyedmohammad, Jammes Manon, Tabasi Abbas, Bach Trung, Ryan Aideen E, Ritter Thomas
Regenerative Medicine Institute, School of Medicine, University of Galway, Galway, Ireland; CURAM Centre for Research in Medical Devices, University of Galway, Galway, Ireland.
Regenerative Medicine Institute, School of Medicine, University of Galway, Galway, Ireland.
Int Immunopharmacol. 2025 Apr 24;153:114480. doi: 10.1016/j.intimp.2025.114480. Epub 2025 Mar 18.
Mesenchymal stromal cell (MSC) apoptosis is essential for their therapeutic effects, including immunomodulation. Previous studies have shown that MSC-derived apoptotic bodies (ApoBDs) also possess immunomodulatory properties. However, compared to small extracellular vesicles, the preparation, characterization, and biological properties of ApoBDs remain underexplored.
ApoBDs were isolated from the conditioned medium of staurosporine-induced apoptotic human MSCs and categorized into large (∼700 nm) and small (∼500 nm) groups. Both types expressed CD90, CD44, and CD73, with low levels of PD-L1, CD11b, and HLA-DR, mirroring their parental MSCs. Functional assays revealed that both ApoBDs inhibited allogeneic T-cell proliferation, with large ApoBDs demonstrating superior efficacy. In macrophage co-culture experiments, both ApoBDs polarized M1 macrophages toward an M2-like phenotype, with large ApoBDs more effectively upregulating CD163 expression. Additionally, both ApoBDs suppressed the proliferation of murine primary T cells. Furthermore, large ApoBDs exhibited enhanced macrophage uptake, as confirmed by flow cytometry and immunocytochemistry. Importantly, no cytotoxicity was observed for either ApoBD type following staurosporine treatment.
Staurosporine-induced ApoBDs are non-cytotoxic and exhibit significant immunomodulatory potential in vitro. Large ApoBDs are more effective than small ApoBDs in T-cell suppression and M2 macrophage polarization, suggesting their potential as an alternative to MSC-based therapies in future studies.
间充质基质细胞（MSC）凋亡对其治疗效果至关重要，包括免疫调节作用。先前的研究表明，MSC来源的凋亡小体（ApoBDs）也具有免疫调节特性。然而，与小细胞外囊泡相比，ApoBDs的制备、表征和生物学特性仍未得到充分研究。
从星形孢菌素诱导凋亡的人MSC的条件培养基中分离出ApoBDs，并将其分为大（约700nm）和小（约500nm）两组。两种类型均表达CD90、CD44和CD73，PD-L1、CD11b和HLA-DR水平较低，与其亲代MSC相似。功能测定表明，两种ApoBDs均抑制同种异体T细胞增殖，大ApoBDs显示出更高的疗效。在巨噬细胞共培养实验中，两种ApoBDs均使M1巨噬细胞极化为M2样表型，大ApoBDs更有效地上调CD163表达。此外，两种ApoBDs均抑制小鼠原代T细胞的增殖。此外，流式细胞术和免疫细胞化学证实，大ApoBDs表现出增强的巨噬细胞摄取能力。重要的是，星形孢菌素处理后，两种类型的ApoBDs均未观察到细胞毒性。
星形孢菌素诱导的ApoBDs无细胞毒性，在体外具有显著的免疫调节潜力。在T细胞抑制和M2巨噬细胞极化方面，大ApoBDs比小ApoBDs更有效，这表明它们在未来研究中作为基于MSC的治疗方法的替代物的潜力。
