ABSTRACT

Purpose

To investigate the immunotherapeutic effects of macrophage-like induced pluripotent stem (iPS) cell-derived suppressor cells (SCs) in ocular immune response and experimental autoimmune uveoretinitis (EAU).

Methods

The genes of Oct3/4, Sox2, Klf4, and c-Myc were transferred to B cells enriched from the spleen cells of C57BL/6 mice by using retrovirus vectors. Transferred B cells were cultured for 17 days to obtain colonies of iPS cells. Through additional steps, iPS-SCs were induced. An antigen-specific T cell proliferation assay was performed with CD4⁺ T cells collected from draining lymph nodes of the mice immunized with human interphotoreceptor retinoid-binding protein (hIRBP) peptide and co-cultured with iPS-SCs. Cytokine concentrations in the culture supernatant were examined. Mice were immunized with hIRBP peptide to induce EAU. The iPS-SCs were administered into the mice one day before the induction of EAU.

Results

The iPS-SCs decreased hIRBP-specific T cell proliferation depending on the number of cells. Productions of tumor necrosis factor-α and interferon-γ were significantly decreased; however, transforming growth factor-β1, nitric oxide, interleukin (IL)-13, IL-17A, and IL-17 F levels were elevated in the supernatant when the collected T cells were co-cultured with iPS-SCs. The iPS-SCs had immunosuppressant effects even without cell-to-cell contact, and their effects were non-specific to the antigen preloaded on iPS-SCs. EAU was significantly milder in the mice administered iPS-SCs prior to immunization.

Conclusions

Macrophage-like iPS-SCs reduced Th1 immune response to a retinal antigen and Th1-mediated EAU in mice. These results showed the possibility of the application of iPS technology to the treatment of noninfectious ocular inflammation, endogenous uveitis, in the future.

中文翻译：

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巨噬细胞样 iPS 衍生抑制细胞降低 Th1 介导的对视网膜抗原的免疫反应

摘要

目的

研究巨噬细胞样诱导的多能干 (iPS) 细胞衍生的抑制细胞 (SCs) 在眼部免疫反应和实验性自身免疫性葡萄膜炎 (EAU) 中的免疫治疗作用。

方法

通过使用逆转录病毒载体将Oct3/4、Sox2、Klf4和c-Myc的基因转移到从C57BL/6小鼠的脾细胞中富集的B细胞中。将转移的B细胞培养17天以获得iPS细胞集落。通过额外的步骤，诱导了 iPS-SC。用从用人光感受器间维甲酸结合蛋白 (hIRBP) 肽免疫并与 iPS-SC 共培养的小鼠的引流淋巴结收集的CD4 ⁺ T 细胞进行抗原特异性 T 细胞增殖测定。检查培养上清液中的细胞因子浓度。用 hIRBP 肽免疫小鼠以诱导 EAU。在诱导 EAU 前一天，将 iPS-SC 施用于小鼠。

结果

iPS-SCs 根据细胞数量减少了 hIRBP 特异性 T 细胞增殖。肿瘤坏死因子-α和干扰素-γ的产生明显减少；然而，当收集的 T 细胞与 iPS-SC 共培养时，上清液中的转化生长因子-β1、一氧化氮、白细胞介素 (IL)-13、IL-17A 和 IL-17 F 水平升高。即使没有细胞间接触，iPS-SCs 也具有免疫抑制作用，并且它们的作用对预载在 iPS-SCs 上的抗原是非特异性的。在免疫前给予 iPS-SC 的小鼠中，EAU 明显更温和。

结论

巨噬细胞样 iPS-SC 降低了小鼠对视网膜​​抗原和 Th1 介导的 EAU 的 Th1 免疫反应。这些结果表明，未来将 iPS 技术应用于治疗非感染性眼部炎症、内源性葡萄膜炎的可能性。