Division of Gastroenterology, Department of Pediatrics, BC's Children's Hospital and Child and Family Research Institute, Vancouver, British Columbia, Canada.
Infect Immun. 2013 Dec;81(12):4649-58. doi: 10.1128/IAI.00655-13. Epub 2013 Oct 7.
Both idiopathic and infectious forms of colitis disrupt normal intestinal epithelial cell (IEC) proliferation and differentiation, although the mechanisms involved remain unclear. Recently, we demonstrated that infection by the attaching and effacing murine pathogen Citrobacter rodentium leads to a significant reduction in colonic goblet cell numbers (goblet cell depletion). This pathology depends on T and/or B cells, as Rag1(-/-) mice do not suffer this depletion during infection, instead suffering high mortality rates. To address the immune mechanisms involved, we reconstituted Rag(-/-) mice with either CD4(+) or CD8(+) T cells. Both T cell subsets increased Rag1(-/-) mouse survival during infection, with mice that received CD8(+) T cells developing colonic ulcers but not goblet cell depletion. In contrast, mice that received CD4(+) T cells showed goblet cell depletion in concert with exaggerated IEC proliferation. To define the possible involvement of T cell-derived cytokines, we infected gamma interferon receptor gene knockout (IFN-γR(-/-)) mice and wild-type mice given interleukin 17A (IL-17A) neutralizing antibodies and found that IFN-γ signaling was required for both goblet cell depletion and increased IEC proliferation. Immunostaining revealed that C. rodentium cells preferentially localized to nonhyperplastic crypts containing numerous goblet cells, whereas hyperplastic, goblet cell-depleted crypts appeared protected from infection. To address whether goblet cell depletion benefits the C. rodentium-infected host, we increased goblet cell numbers using the γ-secretase inhibitor dibenzazepine (DBZ), which resulted in greatly increased pathogen burdens and mortality rates. These results demonstrate that goblet cell depletion reflects host immunomodulation of IEC homeostasis and reflects a novel host defense mechanism against mucosal-adherent pathogens.
特发性和感染性结肠炎均破坏正常的肠道上皮细胞(IEC)增殖和分化,尽管具体机制尚不清楚。最近,我们发现黏附性和侵袭性鼠病原体柠檬酸杆菌感染会导致结肠杯状细胞数量显著减少(杯状细胞耗竭)。这种病理学依赖于 T 和/或 B 细胞,因为 Rag1(-/-) 小鼠在感染期间不会发生这种耗竭,反而会出现高死亡率。为了研究涉及的免疫机制,我们用 CD4(+)或 CD8(+)T 细胞重建 Rag(-/-) 小鼠。这两种 T 细胞亚群都能增加 Rag1(-/-) 小鼠在感染期间的存活率,其中接受 CD8(+)T 细胞的小鼠会出现结肠溃疡,但不会出现杯状细胞耗竭。相比之下,接受 CD4(+)T 细胞的小鼠则出现杯状细胞耗竭,并伴有 IEC 过度增殖。为了明确 T 细胞衍生细胞因子可能的参与作用,我们感染了 γ干扰素受体基因敲除(IFN-γR(-/-))小鼠和给予白细胞介素 17A(IL-17A)中和抗体的野生型小鼠,发现 IFN-γ 信号对杯状细胞耗竭和 IEC 增殖增加都是必需的。免疫染色显示,柠檬酸杆菌细胞优先定位于含有大量杯状细胞的非增生隐窝中,而增生、杯状细胞耗竭的隐窝似乎受到感染的保护。为了确定杯状细胞耗竭是否有利于感染柠檬酸杆菌的宿主,我们用 γ-分泌酶抑制剂二苯并氮杂䓬(DBZ)增加杯状细胞数量,结果导致病原体负荷和死亡率大大增加。这些结果表明,杯状细胞耗竭反映了宿主对 IEC 稳态的免疫调节,反映了一种针对黏附性黏膜病原体的新型宿主防御机制。