Department of Pediatrics, Marcus Center for Pediatric Cellular Therapy, Aflac Cancer & Blood Disorders Center, Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia, USA.
Department of Pediatrics, Marcus Center for Pediatric Cellular Therapy, Aflac Cancer & Blood Disorders Center, Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, Georgia, USA; Division of Biologic and Biomedical Sciences, Laney Graduate School, Emory University Atlanta, Georgia, USA.
Cytotherapy. 2023 Dec;25(12):1277-1284. doi: 10.1016/j.jcyt.2023.08.014. Epub 2023 Oct 10.
Mesenchymal stromal cells (MSCs) are polymorphic, adherent cells with the capability to stimulate tissue regeneration and modulate immunity. MSCs have been broadly investigated for potential therapeutic applications, particularly immunomodulatory properties, wound healing and tissue regeneration. The exact physiologic role of MSCs, however, remains poorly understood, and this gap in knowledge significantly impedes the rational development of therapeutic cells. Here, we considered interferon γ (IFN-γ) and tumor necrosis factor alpha (TNF-α), two cytokines likely encountered physiologically and commonly used in cell manufacturing. For comparison, we studied interleukin-10 (IL-10) (anti-inflammatory) and interleukin-4 (IL-4) (type 2 cytokine).
We directly assessed the effects of these cytokines on bone marrow MSCs by comparing RNA Seq transcriptional profiles. Western blotting and flow cytometry were also used to evaluate effects of cytokine priming.
The type 1 cytokines (IFN-γ and TNF-α) induced striking changes in gene expression and remarkably different profiles from one another. Importantly, priming MSCs with either of these cytokines did not increase variability among multiple donors beyond what is intrinsic to non-primed MSCs from different donors. IFN-γ-primed MSCs expressed IDO1 and chemokines that recruit activated T cells. In contrast, TNF-α-primed MSCs expressed genes in alternate pathways, namely PGE2 and matrix metalloproteinases synthesis, and chemokines that recruit neutrophils. IL-10 and IL-4 priming had little to no effect.
Our data suggest that IFN-γ-primed MSCs may be a more efficacious immunosuppressive therapy aimed at diseases that target T cells (ie, graft-versus-host disease) compared with TNF-α-primed or non-primed MSCs, which may be better suited for therapies in other disease settings. These results contribute to our understanding of MSC bioactivity and suggest rational ex vivo cytokine priming approaches for MSC manufacturing and therapeutic applications.
间充质基质细胞（MSCs）是多形的、贴壁细胞，具有刺激组织再生和调节免疫的能力。MSCs 被广泛研究用于潜在的治疗应用，特别是免疫调节特性、伤口愈合和组织再生。然而，MSCs 的确切生理功能仍知之甚少，这一知识空白严重阻碍了治疗细胞的合理开发。在这里，我们考虑了干扰素γ（IFN-γ）和肿瘤坏死因子α（TNF-α），这两种细胞因子可能在生理上遇到，并常用于细胞制造。为了比较，我们研究了白细胞介素-10（IL-10）（抗炎）和白细胞介素-4（IL-4）（2 型细胞因子）。
我们通过比较 RNA 测序转录谱，直接评估这些细胞因子对骨髓间充质基质细胞的影响。还使用 Western blot 和流式细胞术来评估细胞因子引发的作用。
1 型细胞因子（IFN-γ 和 TNF-α）诱导基因表达发生显著变化，彼此之间的表达谱差异显著。重要的是，用这些细胞因子中的任何一种引发 MSC 不会增加多个供体之间的变异性，超过来自不同供体的未引发 MSC 的固有变异性。IFN-γ 引发的 MSC 表达 IDO1 和招募激活 T 细胞的趋化因子。相比之下，TNF-α 引发的 MSC 表达不同途径的基因，即 PGE2 和基质金属蛋白酶合成以及招募中性粒细胞的趋化因子。IL-10 和 IL-4 引发的作用很小或没有。
我们的数据表明，与 TNF-α 引发或未引发的 MSC 相比，IFN-γ 引发的 MSC 可能是一种更有效的免疫抑制疗法，适用于靶向 T 细胞的疾病（即移植物抗宿主病），而 TNF-α 引发或未引发的 MSC 可能更适合其他疾病情况下的治疗。这些结果有助于我们理解 MSC 的生物活性，并为 MSC 制造和治疗应用提出合理的体外细胞因子引发方法。
