标题：In Vivo Augmentation of Gut-Homing Regulatory T Cell Induction
作者：Bi, Hongzheng; Wasnik, Samiksha; Baylink, David J.; Liu, Chenfan; Tang, Xiaolei
作者机构：[Bi, Hongzheng; Wasnik, Samiksha; Baylink, David J.; Liu, Chenfan; Tang, Xiaolei] Loma Linda Univ, Div Regenerat Med, Dept Med, Loma Linda, CA 92350 U 更多
通讯作者：Tang, XL;Tang, XL
通讯作者地址：[Tang, XL]Loma Linda Univ, Div Regenerat Med, Dept Med, Loma Linda, CA 92350 USA;[Tang, XL]Long Isl Univ, Dept Vet Biomed Sci, Coll Vet Med, Brooklyn, 更多
来源：JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
关键词：Immunology and Infection; gut-homing regulatory T cells;; 25-hydroxyvitamin D 1 alpha-hydroxylase; cytochrome P450 family 27; subfamily B member 1, 1, 25-dihydroxyvitamin D; foxp3; dendritic cells;; retinaldehyde dehydrogenase 2; c-c chemokine receptor type 9
摘要：Inflammatory bowel disease (IBD) is an inflammatory chronic disease in the gastrointestinal tract (GUT). In the United States, there are approximately 1.4 million IBD patients. It is generally accepted that a dysregulated immune response to gut bacteria initiates the disease and disrupts the mucosal epithelial barrier. We recently show that gut-homing regulatory T (Treg) cells are a promising therapy for IBD. Accordingly, this article presents a protocol for in vivo augmentation of gut-homing Treg cell induction. In this protocol, dendritic cells are engineered to produce locally high concentrations of two molecules de novo, active vitamin D (1,25-dihydroxyvitamin D or 1,25[OH](2)D) and active vitamin A (retinoic acid or RA). We chose 1,25(OH)(2)D and RA based on previous findings showing that 1,25(OH)(2)D can induce the expression of regulatory molecules (e.g., forkhead box P3 and interleukin-10) and that RA can stimulate the expression of gut-homing receptors in T cells. To generate such engineered dendritic cells, we use a lentiviral vector to transduce dendritic cells to overexpress two genes. One gene is the cytochrome P450 family 27 subfamily B member 1 that encodes 25-hydroxyvitamin D 1 alpha-hydroxylase, which physiologically catalyzes the synthesis of 1,25(OH)(2)D. The other gene is the aldehyde dehydrogenase 1 family member A2 that encodes retinaldehyde dehydrogenase 2, which physiologically catalyzes the synthesis of RA. This protocol can be used for future investigation of gut-homing Treg cells in vivo.