Edelblum Karen L, Singh Gurminder, Odenwald Matthew A, Lingaraju Amulya, El Bissati Kamal, McLeod Rima, Sperling Anne I, Turner Jerrold R
Department of Pathology, The University of Chicago, Chicago, Illinois.
Department of Ophthalmology and Visual Sciences, The University of Chicago, Chicago, Illinois; Department of Pediatrics, The University of Chicago, Chicago, Illinois.
Gastroenterology. 2015 Jun;148(7):1417-26. doi: 10.1053/j.gastro.2015.02.053. Epub 2015 Mar 4.
BACKGROUND & AIMS: Intraepithelial lymphocytes that express the γδ T-cell receptor (γδ IELs) limit pathogen translocation across the intestinal epithelium by unknown mechanisms. We investigated whether γδ IEL migration and interaction with epithelial cells promote mucosal barrier maintenance during enteric infection.
Salmonella typhimurium or Toxoplasma gondii were administered to knockout (KO) mice lacking either the T cell receptor δ chain (Tcrd) or CD103, or control TcrdEGFP C57BL/6 reporter mice. Intravital microscopy was used to visualize migration of green fluorescent protein (GFP)-tagged γδ T cells within the small intestinal mucosa of mice infected with DsRed-labeled S typhimurium. Mixed bone marrow chimeras were generated to assess the effects of γδ IEL migration on early pathogen invasion and chronic systemic infection.
Morphometric analyses of intravital video microscopy data showed that γδ IELs rapidly localized to and remained near epithelial cells in direct contact with bacteria. Within 1 hour, greater numbers of T gondii or S typhimurium were present within mucosae of mice with migration-defective occludin KO γδ T cells, compared with controls. Pathogen invasion in Tcrd KO mice was quantitatively similar to that in mice with occludin-deficient γδ T cells, whereas invasion in CD103 KO mice, which have increased migration of γδ T cells into the lateral intercellular space, was reduced by 63%. Consistent with a role of γδ T-cell migration in early host defense, systemic salmonellosis developed more rapidly and with greater severity in mice with occludin-deficient γδ IELs, relative to those with wild-type or CD103 KO γδ IELs.
In mice, intraepithelial migration to epithelial cells in contact with pathogens is essential to γδ IEL surveillance and immediate host defense. γδ IEL occludin is required for early surveillance that limits systemic disease.
表达γδ T细胞受体的上皮内淋巴细胞(γδ IELs)通过未知机制限制病原体穿过肠道上皮。我们研究了γδ IEL迁移以及与上皮细胞的相互作用是否在肠道感染期间促进黏膜屏障维持。
将鼠伤寒沙门氏菌或刚地弓形虫接种到缺乏T细胞受体δ链(Tcrd)或CD103的基因敲除(KO)小鼠,或对照TcrdEGFP C57BL/6报告基因小鼠。采用活体显微镜观察感染了DsRed标记的鼠伤寒沙门氏菌的小鼠小肠黏膜内绿色荧光蛋白(GFP)标记的γδ T细胞的迁移。构建混合骨髓嵌合体以评估γδ IEL迁移对早期病原体侵袭和慢性全身感染的影响。
对活体视频显微镜数据的形态计量分析表明,γδ IELs迅速定位并停留在与细菌直接接触的上皮细胞附近。1小时内,与对照相比,迁移缺陷的闭合蛋白KO γδ T细胞的小鼠黏膜内存在更多数量的刚地弓形虫或鼠伤寒沙门氏菌。Tcrd KO小鼠中的病原体侵袭在数量上与闭合蛋白缺陷的γδ T细胞的小鼠相似,而CD103 KO小鼠中γδ T细胞向外侧细胞间隙的迁移增加,其侵袭减少了63%。与γδ T细胞迁移在早期宿主防御中的作用一致,相对于野生型或CD103 KO γδ IELs的小鼠,闭合蛋白缺陷的γδ IELs的小鼠全身沙门氏菌病发展得更快且更严重。
在小鼠中,上皮内迁移至与病原体接触的上皮细胞对于γδ IEL监测和即时宿主防御至关重要。γδ IEL闭合蛋白是限制全身疾病的早期监测所必需的。