School of Pharmaceutical Sciences, The Institute of Immunopharmacology and Immunotherapy, Shandong University, Jinan 250012, China.
Transplantation. 2013 Jun 15;95(11):1324-30. doi: 10.1097/TP.0b013e318290b086.
Foxp3 is a key marker of CD4CD25 regulatory T cells and appears highly specific for regulatory T cells. Human dendritic cells transfected with foxp3 gene also exhibit immunosuppressive functions. We want to understand whether natural killer (NK) cells could be endowed with regulatory properties by transduction of Foxp3 gene.
A recombinant vector (pRV.GFP Foxp3) or control vector (pRV.GFP WWRR) was transferred into NKL/NK-92 cells by an electroporation method. The hFoxp3 gene-modified NK cells were characterized with regard to their proliferation, cytokine production, and cytotoxicity and their regulatory effects on activated human peripheral blood mononuclear cells (hPBMCs) in vitro and trans vivo delayed-type hypersensitivity assay.
We found that the ectopic expression of hFoxp3 in human NK cells resulted in the high production of the immunosuppressive cytokine, interleukin (IL)-10. Luciferase reporter assay showed that the expression of IL-10 is directly regulated by Foxp3. We observed that NKL.Foxp3 cells inhibited the proliferation and activation of phorbol-12-myristate-13-acetate/ionomycin-stimulated hPBMCs; furthermore, NKL.Foxp3 cells significantly suppressed the delayed-type hypersensitivity response, which was induced by anti-CD3 monoclonal antibody-activated hPBMCs. NKL.Foxp3 cell-mediated negative regulatory function was dependent on IL-10 production.
Our findings indicated that NK cells acquired IL-10 phenotype by transduction with foxp3 gene and provided evidence that Foxp3 could exert regulatory function not only in regulatory T cells but also in NK cells. These results suggested that Foxp3 gene-modified NK cells might be potential usefulness on graft-versus-host disease or some autoimmune diseases.
Foxp3 是 CD4+CD25+调节性 T 细胞的关键标志物，似乎高度特异性地存在于调节性 T 细胞中。转染 Foxp3 基因的人树突状细胞也表现出免疫抑制功能。我们想了解是否可以通过转导 Foxp3 基因使自然杀伤 (NK) 细胞获得调节特性。
采用电穿孔法将重组载体（pRV.GFP Foxp3）或对照载体（pRV.GFP WWRR）转染至 NKL/NK-92 细胞。体外和体内迟发型超敏反应试验研究了 hFoxp3 基因修饰的 NK 细胞的增殖、细胞因子产生和细胞毒性及其对活化的人外周血单个核细胞 (hPBMC) 的调节作用。
我们发现人 NK 细胞中 hFoxp3 的异位表达导致免疫抑制细胞因子白细胞介素 (IL)-10 的高表达。荧光素酶报告基因检测显示 IL-10 的表达受 Foxp3 直接调控。我们观察到 NKL.Foxp3 细胞抑制佛波醇 12-肉豆蔻酸 13-乙酸/离子霉素刺激的 hPBMC 的增殖和活化；此外，NKL.Foxp3 细胞显著抑制由抗 CD3 单克隆抗体活化的 hPBMC 诱导的迟发型超敏反应。NKL.Foxp3 细胞介导的负调节功能依赖于 IL-10 的产生。
我们的研究结果表明，NK 细胞通过转导 foxp3 基因获得了 IL-10 表型，并为 Foxp3 不仅在调节性 T 细胞中而且在 NK 细胞中发挥调节功能提供了证据。这些结果表明，Foxp3 基因修饰的 NK 细胞可能在移植物抗宿主病或某些自身免疫性疾病中具有潜在的应用价值。
