TIMP-GLIA (gliadin nanoparticle) treatment in transgenic mice expressing celiac disease associated HLA-DQ8

This submission contains raw data obtained from RNA sequencing of mouse spleen cells. Spleen cells were derived from adult female HLA-DQ8, huCD4 transgenic Ab0 NOD mice, treated intravenously either with tolerogenic, immune-modifying nanoparticles containing gliadin (TIMP-GLIA), particles containing irrelevant control ovalbumin (TIMP-OVA), or not receiving treatment. Mice were then immunized with gliadin (except for negative controls), and spleens were collected after 28-30 days. Spleen cells were stimulated with gliadin (or controls) in culture for 72h, and RNA was obtained from cells for transcriptomic studies. Overall design: 2 groups of female HLA-DQ8, huCD4 transgenic Ab0 NOD mice (Jackson Laboratories) were treated i.v. on days -11 and -3 with TIMP-GLIA, or TIMP-OVA, while other groups remained without treatment (2.5mg/dose; n=11-19, 10-12 weeks of age, raised on gluten-free diet AIN-76A). On day 0, mice from the TIMP-GLIA group, the TIMP-OVA group and a control group of mice (IMMU ONLY) were injected at the base of the tail with 100 µg gliadin in complete Freund's adjuvant, followed by 50 µg gliadin in incomplete Freund's adjuvant on day 14. Another control group (NO TREATMENT) remained untreated/non-immunized. On days 28-30, mice were anesthetized with ketamine/xylazine. Spleens were harvested for analyses. The raw data files for each sample in this submission are not de-multiplexed. All sample metadata are included in the file sample_information.xls linked to to the foot of this record.

本提交数据集包含小鼠脾脏细胞RNA测序所得的原始数据。实验所用脾脏细胞取自成年雌性人白细胞抗原DQ8（HLA-DQ8）阳性、人源CD4（huCD4）转基因Ab0 NOD小鼠，经静脉注射分别给予含麦谷蛋白的免疫耐受型免疫修饰纳米颗粒（TIMP-GLIA）、含无关对照卵清蛋白的纳米颗粒（TIMP-OVA），或不施加任何处理。随后除阴性对照组外，所有小鼠均以麦谷蛋白进行免疫，并于免疫后28至30天采集脾脏。将分离得到的脾脏细胞在体外培养体系中以麦谷蛋白（或对照抗原）刺激72小时，随后提取细胞RNA用于转录组学分析。

实验整体设计如下：将购自杰克逊实验室的雌性人白细胞抗原DQ8（HLA-DQ8）阳性、人源CD4（huCD4）转基因Ab0 NOD小鼠分为2组，分别于第-11天和第-3天经静脉注射TIMP-GLIA或TIMP-OVA，其余组别不施加任何处理（给药剂量为2.5mg/次；每组样本量n=11~19，小鼠周龄为10~12周，饲养于无麸质AIN-76A饲料环境中）。第0天，向TIMP-GLIA组、TIMP-OVA组以及IMMU ONLY对照组小鼠的尾基部注射100微克麦谷蛋白完全弗氏佐剂混悬液，并于第14天注射50微克麦谷蛋白不完全弗氏佐剂混悬液。另有NO TREATMENT对照组全程不接受处理及免疫。于第28至30天，使用氯胺酮/甲苯噻嗪对小鼠实施麻醉，随后采集脾脏用于后续分析。

本提交中所有样本的原始数据文件均未进行解多路复用（de-multiplexed）处理。所有样本元数据均收录于本记录末尾关联的sample_information.xls文件中。