Breitman Maya, Bonfield Tracey L, Caplan Arnold I, Lazarus Hillard M, Haghiac Maricela, LaSalvia Susan, Reese-Koc Jane, Singer Nora G
Case Western Reserve University and MetroHealth Medical Center, Cleveland, Ohio, USA.
Case Western Reserve University, Cleveland, Ohio, USA.
ACR Open Rheumatol. 2022 Feb;4(2):152-160. doi: 10.1002/acr2.11356. Epub 2021 Nov 18.
Seropositive rheumatoid arthritis (RA) is a chronic autoimmune disease that is rarely "cured." Human mesenchymal stem cells (hMSCs) are known to reduce inflammation and restore immune homeostasis. However, methods for predicting therapeutic hMSC potency have not been established. The goal of these studies was to use and refine an ex vivo functional assay that determines potency of hMSCs and can then be validated in clinical trials as a potency measure of hMSCs used therapeutically to treat RA.
Allogeneic hMSCs were cytokine-stimulated, and a conditioned medium (CM) was harvested. The CM was tested for the potential to attenuate RA CD4+ T cell proliferation using suppression assays. Indoleamine 2, 3-dioxygenase (IDO) mRNA, and protein were quantified in hMSCs as a measure to compare hMSCs across (prior) studies.
To mimic a proinflammatory environment that resembles that in RA, interleukin-1(IL1β), tumor necrosis factor α (TNFα), and interferon γ (IFNγ) (alone or in combination) were used to precondition hMSCs. Treating hMSCs with a combination of these cytokines generated a CM "secretome" that suppressed T cell proliferation between 70 and 83%. Forty-eight hours of cytokine preconditioning hMSCs was required to maximize this effect. T cell suppression positively correlated with increases in hMSC cellular IDO mRNA and protein.
By standardizing assays to measure hMSC effects, their potency on T cell suppression can be quantified. These studies demonstrate that hMSCs can be compared functionally to identify optimal preparation(s) for therapeutic use in RA and that the potency of hMSC-dependent T cell suppression may differ between hMSC donors. Clinical studies are warranted to validate the hypothesis that ex vivo potency in suppressing T cells will positively correlate with a reduction in RA disease activity and increase in immunological quiescence.
血清反应阳性类风湿性关节炎（RA）是一种慢性自身免疫性疾病，很少能“治愈”。已知人间充质干细胞（hMSC）可减轻炎症并恢复免疫稳态。然而，尚未建立预测治疗性hMSC效力的方法。这些研究的目的是使用并完善一种体外功能测定法，该方法可确定hMSC的效力，然后在临床试验中作为治疗RA所用hMSC效力的一种衡量标准进行验证。
对同种异体hMSC进行细胞因子刺激，收集条件培养基（CM）。使用抑制试验检测CM减弱RA CD4 + T细胞增殖的潜力。对hMSC中的吲哚胺2,3-双加氧酶（IDO）mRNA和蛋白质进行定量，作为比较（之前）不同研究中hMSC的一种方法。
为模拟类似于RA中的促炎环境，使用白细胞介素-1（IL1β）、肿瘤坏死因子α（TNFα）和干扰素γ（IFNγ）（单独或联合使用）对hMSC进行预处理。用这些细胞因子的组合处理hMSC产生了一种CM“分泌组”，其可抑制70%至83%的T细胞增殖。需要对hMSC进行48小时的细胞因子预处理以使这种效应最大化。T细胞抑制与hMSC细胞IDO mRNA和蛋白质的增加呈正相关。
通过标准化测定hMSC效应的试验，可以量化它们对T细胞抑制的效力。这些研究表明，可以从功能上比较hMSC以确定用于RA治疗的最佳制剂，并且hMSC依赖的T细胞抑制效力在hMSC供体之间可能有所不同。有必要进行临床研究以验证以下假设：体外抑制T细胞的效力将与RA疾病活动度的降低和免疫静止的增加呈正相关。
