CD177 cells produce neutrophil extracellular traps that promote biliary atresia.

BACKGROUND & AIMS: We have previously reported on the potential pathogenic role of neutrophils in biliary atresia (BA). Herein, we aimed to delineate the role of CD177+ neutrophils in the pathogenesis of BA.

METHODS

Immune cells from the livers of mice with rhesus rotavirus-induced BA were analysed. Single-cell RNA-sequencing was performed to specifically analyse Gr-1 (Ly6C/Ly6G) cells in the liver. Gene expression profiles of CD177 cells were analysed using the Smart-Seq RNA-sequencing method, and the pathogenesis of BA was examined in Cd177 mice. Neutrophil extracellular trap (NET) inhibitors were used to determine the role of CD177 cell-derived NETs in BA-associated bile duct damage, and a pilot clinical study evaluated the potential effects of N-acetylcysteine on NET release in BA.

RESULTS

Increased levels of Gr-1 cells were observed in the livers of mice with rhesus rotavirus-induced BA. RNA-sequencing analysis revealed that CD177 cells were the main population of Gr-1 cells and expressed elevated levels of both interferon-stimulated and neutrophil degranulation genes. Cd177 BALB/c mice exhibited delayed disease onset and reduced morbidity and mortality. High numbers of mitochondria were detected in CD177 cells derived from mice with BA; these cells were associated with increased levels of reactive oxygen species and increased NET formation, which induced the apoptosis of biliary epithelial cells in cocultures. In a pilot clinical study, the administration of N-acetylcysteine to patients with BA reduced CD177 cell numbers and reactive oxygen species levels, indicating a potential beneficial effect.

CONCLUSIONS

Our data indicate that CD177 cells play an important role in the initiation of BA pathogenesis via NET formation.

CLINICAL TRIAL REGISTRATION

The pilot study of N-acetylcysteine treatment in patients with BA was registered on the Chinese Clinical Trial Registry (ChiCTR2000040505).

LAY SUMMARY

Neutrophils (a type of innate immune cell, i.e. an immune cell that doesn't target a specific antigen) are thought to play a role in the development of biliary atresia (a rare but potentially lethal condition of the bile ducts that occurs in infants). Herein, we found that neutrophils expressing a particular protein (CD177) played an important role in bile duct damage by releasing a special structure (NET) that can trap and kill pathogens but that can also cause severe tissue damage. A pilot study in patients with biliary atresia showed that inhibiting NETs could have a beneficial effect.