Transplantation Immunology Unit, Departement of Medicine and Diagnostic, Geneva University Hospital, Faculty of Medicine, Geneva, Switzerland.
Division of Neurology, Geneva University Hospital, Faculty of Medicine, Geneva, Switzerland.
Stem Cells Dev. 2020 Jul 1;29(13):853-862. doi: 10.1089/scd.2019.0201. Epub 2020 May 4.
In recent years, great hope has arisen surrounding human stem cells, particularly human induced pluripotent stem (hiPS) cells, as nearly all human tissues can be derived from hiPS cells, using a specific protocol. Therefore, hiPS cells can be a source for replacing defective tissues and make up for the lack of organ donors. However, the alloreactivity of hiPS cells and their derivatives in the context of transplantation remain unclear. Although immunosuppressive drugs can inhibit the T cell compartment, these drugs inhibit partially or not at all natural killer (NK) cells activity. Therefore, the alloreactivity of NK cells against transplanted cells remains to be established. To partially answer this question, we choose, as a model, the potential of cellular therapy for Parkinson's disease (PD). First, we established the in vitro derivation of hiPS cells into mature dopaminergic (mDOPA) neurons, going through an intermediate step called neurosphere (NS) cells. These different cells population were cultured with or without interferon gamma (IFN-γ). They were characterized phenotypically regarding their morphology, and the expression of specific ligands for NK cell receptors expressed by these cells types was investigated. NK cells were isolated from the peripheral blood of healthy donors and cultured in the presence of interleukin 15, to be activated. To test NK cell alloreactivity, a cytotoxic assay was performed with hiPS cells, NS cells, and mDOPA neurons (IFN-γ treated or not) cocultured with allogenic NK cells. Our results show that allogenic NK cells kill hiPS cells (IFN-γ treated or not), but IFN-γ-treated NS cells were protected from killing by allogenic NK cells, compared with untreated NS cells. Finally, mDOPA neurons (IFN-γ treated or not) were partially protected against allogenic NK cell killing. These results indicate that derivatives of hiPS cells, especially NS cells, could be a good product for allogenic transplantation in cellular therapy for PD.
近年来，人们对人类干细胞，特别是人类诱导多能干细胞（hiPS 细胞）寄予厚望，因为几乎所有的人类组织都可以通过特定的方案从 hiPS 细胞中获得。因此，hiPS 细胞可以成为替代缺陷组织和弥补器官捐献者不足的来源。然而，hiPS 细胞及其衍生物在移植中的同种异体反应性尚不清楚。虽然免疫抑制药物可以抑制 T 细胞，但这些药物部分抑制或完全不抑制自然杀伤（NK）细胞的活性。因此，NK 细胞对移植细胞的同种异体反应性仍有待确定。为了部分回答这个问题，我们选择了帕金森病（PD）细胞治疗的潜力作为模型。首先，我们建立了 hiPS 细胞体外分化为成熟多巴胺能（mDOPA）神经元的方法，经历了一个称为神经球（NS）细胞的中间步骤。这些不同的细胞群体在有无干扰素γ（IFN-γ）的情况下进行培养。它们的表型特征是形态学特征，以及这些细胞类型表达的 NK 细胞受体的特异性配体的表达。NK 细胞从健康供体的外周血中分离出来，并在白细胞介素 15 的存在下培养，以被激活。为了测试 NK 细胞的同种异体反应性，将 hiPS 细胞、NS 细胞和 mDOPA 神经元（IFN-γ 处理或不处理）与同种异体 NK 细胞共培养，进行细胞毒性测定。我们的结果表明，同种异体 NK 细胞可以杀死 hiPS 细胞（IFN-γ 处理或不处理），但与未处理的 NS 细胞相比，IFN-γ 处理的 NS 细胞受到同种异体 NK 细胞杀伤的保护。最后，mDOPA 神经元（IFN-γ 处理或不处理）对同种异体 NK 细胞杀伤有一定的保护作用。这些结果表明，hiPS 细胞的衍生物，特别是 NS 细胞，可能是 PD 细胞治疗中同种异体移植的良好产品。
