Jia Haiyan, Moore Melanie, Wadhwa Meenu, Burns Chris
Biotherapeutics and Advanced Therapies, Research and Development, Science and Research Group, Medicines and Healthcare Products Regulatory Agency, Blanche Lane, South Mimms, Potters Bar EN6 3QG, Hertfordshire, UK.
Therapeutic Reference Materials, Standards Lifecycle, Science and Research Group, Medicines and Healthcare Products Regulatory Agency, Blanche Lane, South Mimms, Potters Bar EN6 3QG, Hertfordshire, UK.
Stem Cells Int. 2024 Dec 9;2024:6153235. doi: 10.1155/sci/6153235. eCollection 2024.
Human induced pluripotent stem cell (iPSC)-derived endothelial cells (ECs) have emerged as a promising source of autologous cells with great potential to produce novel cell therapy for ischemic vascular diseases. However, their clinical application still faces numerous challenges including safety concerns such as the potential aberrant immunogenicity derived from the reprogramming process. This study investigated immunological phenotypes of iPSC-ECs by a side-by-side comparison with primary human umbilical vein ECs (HUVECs). Three types of human iPSC-ECs, NIBSC8-EC generated in house and two commercial iPSC-ECs, alongside HUVECs, were examined for surface expression of proteins of immune relevance under resting conditions and after cytokine activation. All iPSC-EC populations failed to express major histocompatibility complex (MHC) Class II on their surface following interferon-gamma (IFN-) treatment but showed similar basal and IFN--stimulated expression levels of MHC Class I of HUVECs. Multiple iPSC-ECs also retained constitutive and tumor necrosis factor-alpha (TNF-)-stimulated expression levels of intercellular adhesion molecule-1 (ICAM-1) like HUVECs. However, TNF- induced a differential expression of E-selectin and vascular cell adhesion molecule-1 (VCAM-1) on iPSC-ECs. Furthermore, real-time monitoring of proliferation of human peripheral blood mononuclear cells (PBMCs) cocultured on an endothelial monolayer over 5 days showed that iPSC-ECs provoked distinct dynamics of PBMC proliferation, which was generally decreased in alloreactivity and IFN--stimulated proliferation of PBMCs compared with HUVECs. Consistently, in the conventional mixed lymphocyte reaction (MLR), the proliferation of total CD3+ and CD4+ T cells after 5-day cocultures with multiple iPSC-EC populations was largely reduced compared to HUVECs. Last, multiple iPSC-EC cocultures secreted lower levels of proinflammatory cytokines than HUVEC cocultures. Collectively, iPSC-ECs manifested many similarities, but also some disparities with a generally weaker inflammatory immune response than primary ECs, indicating that iPSC-ECs may possibly exhibit hypoimmunogenicity corresponding with less risk of immune rejection in a transplant setting, which is important for safe and effective cell therapies.
人诱导多能干细胞（iPSC）来源的内皮细胞（EC）已成为一种很有前景的自体细胞来源，具有为缺血性血管疾病开发新型细胞疗法的巨大潜力。然而，它们的临床应用仍面临诸多挑战，包括安全性问题，如重编程过程可能产生的异常免疫原性。本研究通过与原代人脐静脉内皮细胞（HUVEC）进行并排比较，研究了iPSC-EC的免疫表型。在静息条件下以及细胞因子激活后，对三种类型的人iPSC-EC（在内部生成的NIBSC8-EC和两种商业iPSC-EC）以及HUVEC进行了免疫相关蛋白的表面表达检测。所有iPSC-EC群体在干扰素-γ（IFN-γ）处理后表面均未表达主要组织相容性复合体（MHC）II类，但显示出与HUVEC相似的MHC I类基础表达水平和IFN-γ刺激后的表达水平。多种iPSC-EC也像HUVEC一样保持细胞间黏附分子-1（ICAM-1）的组成性表达水平和肿瘤坏死因子-α（TNF-α）刺激后的表达水平。然而，TNF-α诱导iPSC-EC上E-选择素和血管细胞黏附分子-1（VCAM-1）的差异表达。此外，对在内皮单层上共培养5天的人外周血单个核细胞（PBMC）增殖进行实时监测表明，iPSC-EC引发了PBMC增殖的独特动态，与HUVEC相比，其同种异体反应性和IFN-γ刺激的PBMC增殖通常降低。一致地，在传统的混合淋巴细胞反应（MLR）中，与HUVEC相比，与多种iPSC-EC群体共培养5天后，总CD3+和CD4+ T细胞的增殖大幅减少。最后，多种iPSC-EC共培养分泌的促炎细胞因子水平低于HUVEC共培养。总体而言，iPSC-EC表现出许多相似之处，但也存在一些差异，其炎症免疫反应通常比原代内皮细胞弱，这表明iPSC-EC可能表现出低免疫原性，在移植环境中免疫排斥风险较低，这对于安全有效的细胞治疗很重要。
