Optimization Theory and Applications for Engineering Systems Research Group, Department of Computer Engineering, Excellence Center in Infrastructure Technology and Transportation Engineering, Biomedical Engineering Institute, Chiang Mai University, Chiang Mai, Thailand.
Department of Pediatrics, Case Western Reserve University, Cleveland, OH, USA.
Sci Rep. 2023 Jan 30;13(1):1698. doi: 10.1038/s41598-023-28478-3.
Insights regarding the biodistribution and homing of mesenchymal stromal cells (MSCs), as well as their interaction with alloreactive T-cells are critical for understanding how MSCs can regulate graft-versus-host disease (GVHD) following allogeneic (allo) bone marrow transplantation (BMT). We developed novel assays based on 3D, microscopic, cryo-imaging of whole-mouse-sized volumes to assess the therapeutic potential of human MSCs using an established mouse GVHD model. Following infusion, we quantitatively tracked fluorescently labeled, donor-derived, T-cells and third party MSCs in BMT recipients using multispectral cryo-imaging. Specific MSC homing sites were identified in the marginal zones in the spleen and the lymph nodes, where we believe MSC immunomodulation takes place. The number of MSCs found in spleen of the allo BMT recipients was about 200% more than that observed in the syngeneic group. To more carefully define the effects MSCs had on T cell activation and expansion, we developed novel T-cell proliferation assays including secondary lymphoid organ (SLO) enlargement and Carboxyfluoescein succinimidyl ester (CFSE) dilution. As anticipated, significant SLO volume enlargement and CFSE dilution was observed in allo but not syn BMT recipients due to rapid proliferation and expansion of labeled T-cells. MSC treatment markedly attenuated CFSE dilution and volume enlargement of SLO. These assays confirm evidence of potent, in vivo, immunomodulatory properties of MSC following allo BMT. Our innovative platform includes novel methods for tracking cells of interest as well as assessing therapeutic function of MSCs during GVHD induction. Our results support the use of MSCs treatment or prevention of GVHD and illuminate the wider adoption of MSCs as a standard medicinal cell therapy.
了解间充质基质细胞 (MSCs) 的分布和归巢情况，以及它们与同种异体反应性 T 细胞的相互作用，对于理解 MSCs 如何在同种异体骨髓移植 (alloBMT) 后调节移植物抗宿主病 (GVHD) 至关重要。我们开发了基于 3D 、微观、整个小鼠大小体积冷冻成像的新方法，以使用已建立的小鼠 GVHD 模型评估人 MSCs 的治疗潜力。输注后，我们使用多光谱冷冻成像定量跟踪荧光标记的供体衍生 T 细胞和第三方 MSCs 在 alloBMT 受者中的分布。在脾脏和淋巴结的边缘区确定了特定的 MSC 归巢部位，我们认为 MSC 的免疫调节作用发生在这些部位。在 alloBMT 受者的脾脏中发现的 MSC 数量比同基因组观察到的数量多约 200%。为了更仔细地定义 MSC 对 T 细胞激活和扩增的影响，我们开发了新的 T 细胞增殖测定方法，包括次级淋巴器官 (SLO) 增大和羧基荧光素琥珀酰亚胺酯 (CFSE) 稀释。如预期的那样，由于标记的 T 细胞快速增殖和扩增，在 alloBMT 而不是 synBMT 受者中观察到明显的 SLO 体积增大和 CFSE 稀释。MSC 治疗显著减弱了 CFSE 稀释和 SLO 体积增大。这些测定证实了 alloBMT 后 MSC 具有强大的体内免疫调节特性。我们的创新平台包括用于跟踪感兴趣细胞的新方法，以及评估 GVHD 诱导期间 MSC 治疗功能的新方法。我们的结果支持使用 MSC 治疗或预防 GVHD，并阐明了更广泛地将 MSC 作为标准药物细胞治疗的应用。
