Department of Translational Research and Cellular Therapeutics, Diabetes and Metabolism Research Institute, Beckman Research Institute, City of Hope National Medical Center, Duarte, CA, United States.
Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
Front Immunol. 2020 Apr 28;11:712. doi: 10.3389/fimmu.2020.00712. eCollection 2020.
Type 1 diabetes (T1D) arises secondary to immune-driven destruction of pancreatic β-cells and manifests as insulin-deficient hyperglycemia. We showed that oral vaccination with live attenuated , which simultaneously delivers autoantigens and a TGFβ expression vector to immune cells in the gut mucosa, provides protection against the progression of T1D in non-obese diabetic (NOD) mice. In this study we employed the Sleeping Beauty (SB) transposon system that is composed of a transposase and transposon encoding the td-Tomato to express red fluorescent protein (RFP) to permanently mark the cells that take up the vaccine. After animal vaccination, the transposon labeled-dendritic cells (DCs) with red fluorescence appeared throughout the secondary lymphoid tissues. Furthermore, Sleeping Beauty containing β gene (SB-tgfβ1) co-expressed TGFβ and RFP. The labeled DCs were detected predominantly in Peyer's patches (PP) and mesenteric lymph nodes (MLN) and expressed CD103 surface marker. CD103 DCs induced tolerogenic effects and gut homing. TGFβ significantly increased programmed death-ligand-1 (PDL-1 or CD274) expression in the DCs in the MLN and PP of treated mice. Also, TGFβ increased cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) levels in CD4 cells in MLN and PP. Interestingly, DCs increased in all lymphatic organs of mice vaccinated with oral live -based vaccine expressing preproinsulin (PPI), in combination with TGFβ, IL10, and subtherapeutic-doses of anti-CD3 mAb compared with vehicle-treated mice. These DCs are mostly tolerogenic in MLN and PP. Furthermore the DCs obtained from vaccine-treated but not vehicle-treated mice suppressed T cell proliferation. These data suggest that the MLN and the PP are a central hub for the beneficial anti-diabetic effects of an oral -based vaccine prevention of diabetes in rodents.
1 型糖尿病（T1D）是由胰腺β细胞的免疫驱动破坏引起的，并表现为胰岛素缺乏性高血糖。我们表明，用活减毒疫苗进行口服接种，该疫苗同时将自身抗原和 TGFβ表达载体递送到肠道黏膜中的免疫细胞，可提供对非肥胖型糖尿病（NOD）小鼠 T1D 进展的保护。在这项研究中，我们采用了 Sleeping Beauty（SB）转座子系统，该系统由转座酶和编码 td-Tomato 的转座子组成，用于表达红色荧光蛋白（RFP），以永久标记摄取疫苗的细胞。动物接种疫苗后，带有红色荧光的转座子标记树突状细胞（DC）出现在次级淋巴组织中。此外，含有β基因的 Sleeping Beauty（SB-tgfβ1）共表达 TGFβ和 RFP。标记的 DC 主要在派尔氏集合淋巴结（PP）和肠系膜淋巴结（MLN）中检测到，并表达 CD103 表面标记物。CD103 DC 诱导耐受和肠道归巢。TGFβ显著增加了治疗小鼠 MLN 和 PP 中 DC 的程序性死亡配体-1（PDL-1 或 CD274）表达。此外，TGFβ增加了 MLN 和 PP 中 CD4 细胞中的细胞毒性 T 淋巴细胞相关蛋白-4（CTLA-4）水平。有趣的是，与对照处理的小鼠相比，用口服活疫苗（表达前胰岛素原（PPI））接种的小鼠的所有淋巴器官中均增加了 DC，该疫苗与 TGFβ、IL10 和亚治疗剂量的抗-CD3 mAb 联合使用。与对照处理的小鼠相比，这些 MLN 和 PP 中的 DC 大多具有耐受性。此外，来自疫苗处理而不是对照处理的小鼠的 DC 抑制了 T 细胞增殖。这些数据表明，MLN 和 PP 是口服疫苗预防啮齿动物糖尿病的有益抗糖尿病作用的中心枢纽。
