Immunogenetics and Histocompatibility Laboratory, Department of Genetics, Universidade Federal do Paraná (UFPR), Curitiba 81530-001, PR, Brazil.
Core for Cell Technology, School of Medicine, Pontifícia Universidade Católica do Paraná (PUCPR), Curitiba 80910-215, PR, Brazil.
Cells. 2022 Apr 14;11(8):1339. doi: 10.3390/cells11081339.
Mesenchymal stromal cells (MSCs) have been used in immunosuppressive therapy due to their therapeutic effects, with the HLA-G molecule seeming to play a fundamental role. This work evaluated alternative MSC sources to bone marrow (BM), namely, umbilical cord tissue (UC), adipose tissue (AD) and dental pulp tissue (DP), and the influence of interferon-γ (IFN-γ) and hypoxia on the cultivation of these cells for use in immunosuppression therapies. Expression of costimulatory markers CD40, CD80 and CD86 and immunosuppressive molecules CD152 and HLA-G was analyzed. Lymphocyte inhibition assays were also performed. Sequencing of the HLA-G gene from exons 1 to 5 was performed using next-generation sequencing to determine the presence of alleles. UC-derived MSCs (UCMSCs) expressed higher CD152 and HLA-G1 under standard cultivation. UCMSCs and DP-derived MSCs (DPSCs) secreted similar levels of HLA-G5. All MSC sources inhibited the proliferation of peripheral blood mononuclear cells (PBMCs); growth under regular versus hypoxic conditions resulted in similar levels of inhibition. When IFN-γ was added, PBMC growth was inhibited to a lesser extent by UCMSCs. The HLA-G*01:04:01:01 allele appears to generate a more efficient MSC response in inhibiting lymphocyte proliferation. However, the strength of this conclusion was limited by the small sample size. UCMSCs are an excellent alternative to BM in immunosuppressive therapy: they express high concentrations of inhibitory molecules and can be cultivated without stimuli, which minimizes cost.
间充质基质细胞（MSCs）因其治疗效果而被用于免疫抑制治疗，其中 HLA-G 分子似乎起着重要作用。本工作评估了骨髓（BM）以外的 MSCs 替代来源，即脐带组织（UC）、脂肪组织（AD）和牙髓组织（DP），以及干扰素-γ（IFN-γ）和低氧对这些细胞培养的影响，以用于免疫抑制治疗。分析了共刺激分子 CD40、CD80 和 CD86 以及免疫抑制分子 CD152 和 HLA-G 的表达。还进行了淋巴细胞抑制测定。使用下一代测序对 HLA-G 基因的外显子 1 到 5 进行测序，以确定等位基因的存在。在标准培养条件下，UC 来源的 MSC（UCMSCs）表达更高水平的 CD152 和 HLA-G1。UCMSCs 和 DP 来源的 MSC（DPSCs）分泌相似水平的 HLA-G5。所有 MSC 来源均抑制外周血单个核细胞（PBMCs）的增殖；在常氧和低氧条件下生长导致相似的抑制水平。当添加 IFN-γ 时，UCMSCs 对 PBMC 生长的抑制作用较小。HLA-G*01:04:01:01 等位基因似乎在抑制淋巴细胞增殖方面产生更有效的 MSC 反应。然而，由于样本量小，这一结论的强度受到限制。UCMSCs 是免疫抑制治疗中 BM 的极佳替代物：它们表达高浓度的抑制分子，并且可以在没有刺激的情况下培养，从而最大限度地降低成本。
