Department of Pediatric and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45229, USA.
J Surg Res. 2013 Sep;184(1):322-9. doi: 10.1016/j.jss.2013.05.032. Epub 2013 Jun 1.
Biliary atresia (BA) is a unique neonatal disease resulting from inflammatory and fibrosing obstruction of the extrahepatic biliary tree. Previous studies have demonstrated the critical role of innate immunity and the Th1 response to activated inflammatory cells and overexpressed cytokines in the pathogenesis of BA. Myeloid differentiation factor 88 (MyD88) is a critical adaptor molecule that has been shown to play a crucial role in immunity. We investigated the role of MyD88 in the inflammatory response and development of cholangiopathy in murine BA.
MyD88 knockout (MyD88(-/-)) and wild-type (WT) BALB/c pups were injected with Rhesus rotavirus or saline on day 1 of life. The mice were monitored for clinical symptoms of BA, including jaundice, acholic stools, bilirubinuria, and death. The liver and extrahepatic bile ducts were harvested for histologic evaluation and the quantification of viral content, determination of cytokine expression, and detection of inflammatory cells.
Rhesus rotavirus infection produced symptoms in 100% of both MyD88(-/-) and WT pups, with survival of 18% of WT and 0% of MyD88(-/-) mice. Histologic analysis demonstrated bile duct obstruction in both MyD88(-/-) and WT mice. Viral titers obtained 7 d after infection and expression of interferon-γ and tumor necrosis factor-α at day 3, 5, 8, and 12 after infection revealed no significant differences between the WT and MyD88(-/-) mice. Flow cytometry demonstrated similar levels of activated CD8+ T cells and natural killer cells.
The pathogenesis of murine BA is independent of the MyD88 signaling inflammatory pathway, suggesting alternative mechanisms are crucial in the induction of the model.
先天性胆道闭锁(BA)是一种独特的新生儿疾病,由肝外胆管的炎症和纤维化阻塞引起。先前的研究表明,固有免疫和 Th1 反应对激活的炎症细胞和过度表达的细胞因子在 BA 的发病机制中起着关键作用。髓样分化因子 88(MyD88)是一种关键的衔接分子,已被证明在免疫中起着至关重要的作用。我们研究了 MyD88 在鼠 BA 的炎症反应和胆管病发展中的作用。
在出生后第 1 天,用恒河猴轮状病毒或生理盐水注射 MyD88 敲除(MyD88(-/-))和野生型(WT)BALB/c 幼仔。监测 BA 的临床症状,包括黄疸、白陶土样粪便、胆红素尿和死亡。采集肝脏和肝外胆管进行组织学评估和病毒含量的定量、细胞因子表达的测定以及炎症细胞的检测。
恒河猴轮状病毒感染使 100%的 MyD88(-/-)和 WT 幼仔出现症状,WT 幼仔的存活率为 18%,而 MyD88(-/-)幼仔的存活率为 0%。组织学分析表明,MyD88(-/-)和 WT 小鼠均存在胆管阻塞。感染后 7 天获得的病毒滴度和感染后第 3、5、8 和 12 天 IFN-γ和肿瘤坏死因子-α的表达显示 WT 和 MyD88(-/-) 小鼠之间无显著差异。流式细胞术显示激活的 CD8+T 细胞和自然杀伤细胞水平相似。
鼠 BA 的发病机制独立于 MyD88 信号炎症途径,表明诱导该模型的替代机制至关重要。