a Division of Molecular Medicine , Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University , Bangkok , Thailand.
b Graduate Program in Immunology , Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University , Bangkok , Thailand.
Hum Vaccin Immunother. 2018 Jun 3;14(6):1423-1431. doi: 10.1080/21645515.2018.1431598. Epub 2018 Feb 21.
Tumor escapes host immune responses by producing immunosuppressive cytokines, such as IL-10 and TGF-β, secreted into the tumor microenvironment. These cytokines play important roles in the suppression of dendritic cell (DC) function, leading to decreased immune responses of the effector CD4 and CD8 T cells. To improve DC functions and enhance cytolytic activity of activated effector T-cells, we suppressed the effect of these cytokines on DCs by using specific neutralizing antibodies that inhibit IL-10 and TGF-β receptors. Monocyte-derived DCs generated in vitro showed up-regulation of MHC (HLA-DR) and co-stimulatory molecules (CD40 and CD86). The IL-10 and TGF-β receptors were expressed and localized on cell membrane of DCs, as shown by Western blot analysis and immunofluorescence staining, whereas the IL-10 and TGF-β ligands were detected in the culture supernatants of DCs and cholangiocarcinoma (CCA) cell line, respectively. Inhibition of the IL-10 and TGF-β receptors on DCs by specific neutralizing antibodies significantly increased level of IFN-γ and enhanced cytolytic activity of the DC-activated effector T-cells against CCA cell line. These results indicate that the IL-10 and TGF-β receptors are the targets for inhibition to increase DC functions and enhance cytolytic activity of the DC-activated effector T-cells against CCA cells. Thus, inhibition of the IL-10 and TGF-β receptors on DCs is crucial in the preparation of DC-activated effector T cells for adoptive T-cell therapy.
肿瘤通过产生免疫抑制细胞因子（如 IL-10 和 TGF-β）来逃避宿主的免疫反应，这些细胞因子分泌到肿瘤微环境中。这些细胞因子在抑制树突状细胞（DC）功能方面发挥着重要作用，导致效应 CD4 和 CD8 T 细胞的免疫反应减弱。为了改善 DC 功能并增强激活效应 T 细胞的细胞毒性活性，我们使用特异性中和抗体抑制 IL-10 和 TGF-β 受体，从而抑制这些细胞因子对 DC 的作用。体外生成的单核细胞来源的 DC 表现出 MHC（HLA-DR）和共刺激分子（CD40 和 CD86）的上调。Western blot 分析和免疫荧光染色显示，IL-10 和 TGF-β 受体表达并定位于 DC 的细胞膜上，而 IL-10 和 TGF-β 配体分别在 DC 和胆管癌细胞系的培养上清液中被检测到。特异性中和抗体对 DC 上的 IL-10 和 TGF-β 受体的抑制显著增加了 IFN-γ 的水平，并增强了 DC 激活的效应 T 细胞对胆管癌细胞系的细胞毒性活性。这些结果表明，IL-10 和 TGF-β 受体是抑制的靶点，可增加 DC 功能并增强 DC 激活的效应 T 细胞对胆管癌细胞的细胞毒性活性。因此，抑制 DC 上的 IL-10 和 TGF-β 受体对于制备用于过继性 T 细胞治疗的 DC 激活效应 T 细胞至关重要。
