Cross-activating invariant NKT cells and kupffer cells suppress cholestatic liver injury in a mouse model of biliary obstruction.

Abstract

Both Kupffer cells and invariant natural killer T (iNKT) cells suppress neutrophil-dependent liver injury in a mouse model of biliary obstruction. We hypothesize that these roles are interdependent and require iNKT cell-Kupffer cell cross-activation. Female, wild-type and iNKT cell-deficient C57Bl/6 mice were injected with magnetic beads 3 days prior to bile duct ligation (BDL) in order to facilitate subsequent Kupffer cell isolation. On day three post-BDL, the animals were euthanized and the livers dissected. Necrosis was scored; Kupffer cells were isolated and cell surface marker expression (flow cytometry), mRNA expression (qtPCR), nitric oxide (NO (.) ) production (Griess reaction), and protein secretion (cytometric bead-array or ELISAs) were determined. To address the potential role of NO (.) in suppressing neutrophil accumulation, a group of WT mice received 1400W, a specific inducible nitric oxide synthase (iNOS) inhibitor, prior to BDL. To clarify the mechanisms underlying Kupffer cell-iNKT cell cross-activation, WT animals were administered anti-IFN-γ or anti-lymphocyte function-associated antigen (LFA)-1 antibody prior to BDL. Compared to their WT counterparts, Kupffer cells obtained from BDL iNKT cell-deficient mice expressed lower iNOS mRNA levels, produced less NO (.) , and secreted more neutrophil chemoattractants. Both iNOS inhibition and IFN-γ neutralization increased neutrophil accumulation in the livers of BDL WT mice. Anti-LFA-1 pre-treatment reduced iNKT cell accumulation in these same animals. These data indicate that the LFA-1-dependent cross-activation of iNKT cells and Kupffer cells inhibits neutrophil accumulation and cholestatic liver injury.

• Authors: Caroline C Duwaerts, Eric P Sun, Chao-Wen Cheng, Nico van Rooijen, Stephen H Gregory
• Journal: PloS one (PLoS One) Vol. 8 Issue 11 Pg. e79702 ( 2013) ISSN: 1932-6203 [Electronic] United States
• PMID: 24260285 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Nitric Oxide Synthase Type II

Topics

• Animals
• Cholestasis (immunology, metabolism)
• Disease Models, Animal
• Female
• Flow Cytometry
• Kupffer Cells (immunology, metabolism)
• Liver (immunology, injuries, metabolism)
• Lymphocyte Activation (immunology, physiology)
• Mice
• Mice, Inbred C57BL
• Natural Killer T-Cells (immunology, metabolism)
• Nitric Oxide Synthase Type II (genetics, metabolism)
• Real-Time Polymerase Chain Reaction