Walther Ashley, Mohanty Sujit K, Donnelly Bryan, Coots Abigail, Lages Celine S, Lobeck Inna, Dupree Phylicia, Meller Jaroslaw, McNeal Monica, Sestak Karol, Tiao Greg
Department of Pediatric and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio;
Division of Gastroenterology, Hepatology, and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio;
Am J Physiol Gastrointest Liver Physiol. 2015 Sep 15;309(6):G466-74. doi: 10.1152/ajpgi.00079.2015. Epub 2015 Jul 23.
Biliary atresia (BA), a neonatal obstructive cholangiopathy, remains the most common indication for pediatric liver transplantation in the United States. In the murine model of BA, Rhesus rotavirus (RRV) VP4 surface protein determines biliary duct tropism. In this study, we investigated how VP4 governs induction of murine BA. Newborn mice were injected with 16 strains of rotavirus and observed for clinical symptoms of BA and mortality. Cholangiograms were performed to confirm bile duct obstruction. Livers and bile ducts were harvested 7 days postinfection for virus titers and histology. Flow cytometry assessed mononuclear cell activation in harvested cell populations from the liver. Cytotoxic NK cell activity was determined by the ability of NK cells to kill noninfected cholangiocytes. Of the 16 strains investigated, the 6 with the highest homology to the RRV VP4 (>87%) were capable of infecting bile ducts in vivo. Although the strain Ro1845 replicated to a titer similar to RRV in vivo, it caused no symptoms or mortality. A Ro1845 reassortant containing the RRV VP4 induced all BA symptoms, with a mortality rate of 89%. Flow cytometry revealed that NK cell activation was significantly increased in the disease-inducing strains and these NK cells demonstrated a significantly higher percentage of cytotoxicity against noninfected cholangiocytes. Rotavirus strains with >87% homology to RRV's VP4 were capable of infecting murine bile ducts in vivo. Development of murine BA was mediated by RRV VP4-specific activation of mononuclear cells, independent of viral titers.
胆道闭锁(BA)是一种新生儿梗阻性胆管病,在美国仍然是儿童肝移植最常见的适应证。在BA的小鼠模型中,恒河猴轮状病毒(RRV)VP4表面蛋白决定胆管嗜性。在本研究中,我们调查了VP4如何引发小鼠BA。给新生小鼠注射16种轮状病毒株,并观察BA的临床症状和死亡率。进行胆管造影以确认胆管梗阻。感染后7天收获肝脏和胆管,用于检测病毒滴度和组织学检查。流式细胞术评估从肝脏收获的细胞群体中的单核细胞活化情况。细胞毒性NK细胞活性通过NK细胞杀伤未感染胆管细胞的能力来确定。在所研究的16种毒株中,与RRV VP4同源性最高(>87%)的6种能够在体内感染胆管。尽管Ro1845毒株在体内的复制滴度与RRV相似,但它未引起任何症状或死亡。含有RRV VP4的Ro1845重配株引发了所有BA症状,死亡率为89%。流式细胞术显示,在致病毒株中NK细胞活化显著增加,并且这些NK细胞对未感染胆管细胞的细胞毒性百分比显著更高。与RRV的VP4同源性>87%的轮状病毒株能够在体内感染小鼠胆管。小鼠BA的发生是由RRV VP4特异性激活单核细胞介导的,与病毒滴度无关。