Expression data from WT and AIMp1KO mouse bone marrow-derived dendritic cells

Dendritic cells are critical regulators of adaptive immune responses which can process and present antigens to effector cells. As a structural component of multi-enzyme aminoacyl tRNA synthetase complex, AIMp1 (p43) is identified to promote type 1 T-cell responses by DCs and play important roles in both antitumor and antiviral immunity. We use high-throughput microarray analysis to compare the gene expression profiles of WT vs. AIMp1KO bone marrow-derived DCs (BMDC) to study the mechanism by which DC-expressed AIMp1 promote type 1 T-cell immune responses. WT or AIMp1KO BMDCs were loaded with antigens and matured for RNA extraction and hybridization on Affymetrix microarrays. Cells were unloaded or loaded with OVA protein and SIINFEKL peptide (10ug/mL) for 3 hours in media containing 5% serum; then matured with cytokine cocktail (10ng/mL TNFa, 15ng/mL IL-6, 10ng/mL IL-1b, 1ug/mL PGE2) for 48hours prior to harvest.

树突状细胞（Dendritic cells, DC）是适应性免疫应答的关键调控因子，可加工并提呈抗原给效应免疫细胞。作为多酶氨酰tRNA合成酶复合物的结构组分，AIMp1（p43）已被证实可通过树突状细胞促进1型T细胞应答，在抗肿瘤与抗病毒免疫中均发挥重要作用。本研究采用高通量微阵列分析技术，对比野生型（Wild Type, WT）与AIMp1基因敲除（AIMp1KO）骨髓来源树突状细胞（bone marrow-derived DCs, BMDC）的基因表达谱，以探究DC表达的AIMp1促进1型T细胞免疫应答的分子机制。将野生型或AIMp1KO BMDC负载抗原并诱导成熟后，进行RNA提取，并在Affymetrix微阵列上完成杂交。实验中，细胞分为未负载抗原组与负载抗原组：负载组采用卵清蛋白（Ovalbumin, OVA）蛋白与SIINFEKL肽（10μg/mL）在含5%血清的培养基中孵育3小时；随后使用细胞因子混合液（10ng/mL 肿瘤坏死因子α（TNFα）、15ng/mL 白细胞介素6（IL-6）、10ng/mL 白细胞介素1β（IL-1β）、1μg/mL 前列腺素E2（PGE2））诱导细胞成熟48小时，最后收集细胞。