Effector role of neonatal hepatic CD8+ lymphocytes in epithelial injury and autoimmunity in experimental biliary atresia
Effector role of neonatal hepatic CD8+ lymphocytes in epithelial injury and autoimmunity in experimental biliary atresia
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DOI：
10.1053/j.gastro.2007.04.031
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发表时间：
2007-07-01
期刊：
影响因子：
29.4
通讯作者：
Bezerra, Jorge A.
中科院分区：
文献类型：
--
作者：
Shivakumar, Pranavkumar;Sabla, Gregg;Bezerra, Jorge A.
背景与目的：淋巴细胞存在于胆道闭锁婴儿的肝脏中，但新生儿淋巴细胞是否调节疾病的发病机制尚不清楚。在此，我们利用胆道闭锁实验模型，通过研究T淋巴细胞在新生小鼠肝外胆管破坏中的作用来探讨这一问题。
方法：用恒河猴轮状病毒接种新生小鼠，然后采用多色流式细胞术量化肝淋巴细胞干扰素 -γ的表达，并通过实时聚合酶链反应检测促炎细胞因子的mRNA表达。接着确定抗体介导的淋巴细胞亚型耗竭对胆道梗阻发展的影响，以及通过共培养和细胞转移实验研究淋巴细胞亚型在新生胆道疾病中的效应作用。
结果：轮状病毒感染导致新生肝T细胞干扰素 -γ过度表达。在这些细胞中，CD4 +细胞的耗竭并未改变新生胆管炎症损伤和梗阻的进程。相反，CD8 +细胞的缺失显著抑制了胆管损伤，防止了管腔梗阻，并恢复了胆汁流动。共培养实验表明，经轮状病毒激活但非未激活的CD8 +细胞对胆管细胞具有细胞毒性。在过继转移实验中，我们发现激活的CD8 +细胞优先归巢到新生小鼠的肝外胆管并侵入其上皮内层。
结论：激活的新生CD8 +细胞能够激活促炎程序，靶向病变和健康的胆管上皮，驱动胆道闭锁的表型表达，因此构成了阻止疾病进展的潜在治疗靶点。
Background & Aims: Lymphocytes populate the livers of infants with biliary atresia, but it is unknown whether neonatal lymphocytes regulate pathogenesis of disease. Here, we investigate this question by examining the role of T lymphocytes in the destruction of extrahepatic bile ducts of neonatal mice using an experimental model of biliary atresia. Methods: Inoculation of neonatal mice with rhesus rotavirus followed by multistaining flow cytometry to quantify expression of interferon-gamma by hepatic lymphocytes, and real-time polymerase chain reaction for mRNA expression of pro-inflammatory cytokines. This was followed by determining the consequences of antibody-mediated depletion of lymphocyte subtypes on the development of biliary obstruction, and coculture and cell transfer experiments to investigate the effector role of lymphocyte subtypes on neonatal biliary disease. Results: Rotavirus infection results in overexpression of interferon-gamma by neonatal hepatic T cells. Among these cells, depletion of CD4+ cells did not change the course of inflammatory injury and obstruction of neonatal bile ducts. In contrast, loss of CD8(+) cells remarkably suppressed duct injury, prevented luminal obstruction, and restored bile flow. Coculture experiments showed that rotavirus-primed, but not naive, CD8(+) cells were cytotoxic to cholangiocytes. In adoptive transfer experiments, we found that primed CD8(+) cells preferentially homed to extrahepatic bile ducts of neonatal mice and invaded their epithelial lining. Conclusions: Primed neonatal CD8(+) cells can activate a pro-inflammatory program, target diseased and healthy duct epithelium, and drive the phenotypic expression of biliary atresia, thus constituting a potential therapeutic target to halt disease progression.