Kennedy Institute of Rheumatology, University of Oxford, Oxford, UK.
School of Infection and Immunity, University of Glasgow, Glasgow, UK.
Curr Protoc. 2024 Jul;4(7):e1092. doi: 10.1002/cpz1.1092.
The intestinal inflammation induced by injection of naïve CD4 T cells into lymphocyte-deficient hosts (more commonly known as the T cell transfer model of colitis) shares many features of idiopathic inflammatory bowel disease (IBD) in humans, such as epithelial cell hyperplasia, crypt abscess formation, and dense lamina propria lymphocyte infiltration. As such, it provides a useful tool for studying mucosal immune regulation as it relates to the pathogenesis and treatment of IBD in humans. In the IBD model described here, colitis is induced in Rag (recombination-activating gene)-deficient mice by reconstitution of these mice with naïve CD4CD45RB T cells through adoptive T cell transfer. Although different recipient hosts of cell transfer can be used, Rag-deficient mice are the best characterized and support studies that are both flexible and reproduceable. As described in the Basic Protocol, in most studies the transferred cells consist of naïve CD4 T cells (CD45RB T cells) derived by fluorescence-activated cell sorting from total CD4 T cells previously purified using immunomagnetic negative selection beads. In a Support Protocol, methods to characterize colonic disease progression are described, including the monitoring of weight loss and diarrhea and the histological assessment of colon pathology. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Induction of IBD in Rag-deficient mice by the transfer of naïve CD4CD45RB T cells Support Protocol: Monitoring development of colitis.
注射幼稚 CD4 T 细胞到淋巴细胞缺陷宿主(通常称为结肠炎 T 细胞转移模型)引起的肠道炎症与人类特发性炎症性肠病(IBD)具有许多共同特征,如上皮细胞增生、隐窝脓肿形成和固有层淋巴细胞浸润。因此,它为研究与人类 IBD 发病机制和治疗相关的黏膜免疫调节提供了有用的工具。在本文描述的 IBD 模型中,通过幼稚 CD4CD45RB T 细胞的过继性 T 细胞转移来重建 Rag(重组激活基因)缺陷小鼠,诱导结肠炎。尽管可以使用不同的细胞转移受体宿主,但 Rag 缺陷小鼠的特征最明显,支持灵活且可重现的研究。如基本方案所述,在大多数研究中,转移的细胞包括幼稚 CD4 T 细胞(CD45RB T 细胞),通过荧光激活细胞分选从先前使用免疫磁珠阴性选择珠纯化的总 CD4 T 细胞中获得。在一个支持方案中,描述了监测结肠炎进展的方法,包括体重减轻和腹泻的监测以及结肠病理的组织学评估。© 2024 作者。Wiley Periodicals LLC 出版的当前方案。基本方案:幼稚 CD4CD45RB T 细胞过继转移诱导 Rag 缺陷型小鼠 IBD 支持方案:监测结肠炎的发展。