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Macrophages are targeted by rotavirus in experimental biliary atresia and induce neutrophil chemotaxis by Mip2/Cxcl2
Sujit K Mohanty1, Cláudia A P Ivantes, Reena Mourya
1Cincinnati Children's Hospital Medical Center and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229, USA.
Abstract:
Biliary atresia is an obstructive cholangiopathy of unknown etiology. Although the adaptive immune system has been shown to regulate the obstruction of bile ducts in a rotavirus-induced mouse model, little is known about the virus-induced inflammatory response. Here, we hypothesized that cholangiocytes secrete chemoattractants in response to rotavirus. To test this hypothesis, we infected cholangiocyte and macrophage cell lines with rhesus rotavirus type A (RRV), quantified cytokines and chemokines and measured the migration of splenocytes. We also used PCR and immunostaining to search for new cellular targets of RRV in the liver. We found that RRV-infected cholangiocytes induced the mRNA expression for chemokines, but conditioned media failed to promote chemotaxis of splenocytes. Analyzing livers after viral challenge, we detected RRV in hepatic macrophages and demonstrated that media from RRV-infected macrophages have high concentrations of cytokines and chemokines and induced chemotaxis of neutrophils. Most notably, addition of anti-Mip2/Cxcl2 antibodies depleted this chemokine in the conditioned media and completely prevented neutrophil chemotaxis. In conclusion, infected cholangiocytes did not promote chemotaxis of inflammatory cells. Investigating alternate cellular targets of RRV, we detected the virus in hepatic macrophages and found that infected macrophages promoted neutrophil chemotaxis by release of Mip2/Cxcl2 in response to RRV.
Insights
Rotavirus infection of liver cells does not directly cause inflammation. Instead, infected hepatic macrophages release chemokines that attract neutrophils, contributing to biliary atresia pathogenesis.
Area of Science:
- Hepatology
- Virology
- Immunology
Background:
- Biliary atresia is an obstructive cholangiopathy with unknown causes.
- The role of virus-induced inflammation in biliary atresia is not well understood.
- Previous studies suggested adaptive immune involvement in a rotavirus mouse model.
Purpose of the Study:
- To investigate the role of cholangiocytes and hepatic macrophages in rotavirus-induced inflammation.
- To identify cellular targets of rotavirus in the liver and their inflammatory responses.
- To determine the specific mechanisms by which rotavirus triggers inflammatory cell recruitment.
Main Methods:
- Infection of cholangiocyte and macrophage cell lines with rhesus rotavirus type A (RRV).
- Quantification of cytokines and chemokines using PCR and protein assays.
- Assessment of splenocyte and neutrophil migration in response to conditioned media.
- Detection of RRV in liver tissues using PCR and immunostaining.
- Inhibition of neutrophil chemotaxis using anti-Mip2/Cxcl2 antibodies.
Main Results:
- RRV-infected cholangiocytes upregulated chemokine mRNA but did not induce splenocyte migration.
- RRV was detected in hepatic macrophages within infected liver tissues.
- Conditioned media from RRV-infected macrophages showed high cytokine and chemokine concentrations.
- RRV-infected macrophages induced neutrophil chemotaxis, which was blocked by anti-Mip2/Cxcl2 antibodies.
Conclusions:
- Cholangiocytes infected with RRV do not directly promote inflammatory cell chemotaxis.
- Hepatic macrophages are a cellular target for RRV in the liver.
- Infected hepatic macrophages recruit neutrophils via Mip2/Cxcl2 (CXCL2) chemokine release, contributing to biliary atresia.
- Targeting Mip2/Cxcl2 may offer a therapeutic strategy for rotavirus-induced biliary inflammation.
