Cholangiocyte expression of alpha2beta1-integrin confers susceptibility to rotavirus-induced experimental biliary atresia.
Inoculation of BALB/c mice with rhesus rotavirus (RRV) in the newborn period results in biliary epithelial cell (cholangiocyte) infection and the murine model of biliary atresia. Rotavirus infection of a cell requires attachment, which is governed in part by cell-surface expression of integrins such as alpha2beta1. We hypothesized that cholangiocytes were susceptible to RRV infection because they express alpha2beta1. RRV attachment and replication was measured in cell lines derived from cholangiocytes and hepatocytes. Flow cytometry was performed on these cell lines to determine whether alpha2beta1 was present. Cholangiocytes were blocked with natural ligands, a monoclonal antibody, or small interfering RNA against the alpha2-subunit and were infected with RRV. The extrahepatic biliary tract of newborn mice was screened for the expression of the alpha2beta1-integrin. Newborn mice were pretreated with a monoclonal antibody against the alpha2-subunit and were inoculated with RRV. RRV attached and replicated significantly better in cholangiocytes than in hepatocytes. Cholangiocytes, but not hepatocytes, expressed alpha2beta1 in vitro and in vivo. Blocking assays led to a significant reduction in attachment and yield of virus in RRV-infected cholangiocytes. Pretreatment of newborn pups with an anti-alpha2 monoclonal antibody reduced the ability of RRV to cause biliary atresia in mice. Cell-surface expression of the alpha2beta1-integrin plays a role in the mechanism that confers cholangiocyte susceptibility to RRV infection.
Jafri
Mubeen
M
Department of Pediatric and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Donnelly
Bryan
B
Allen
Steven
S
Bondoc
Alex
A
McNeal
Monica
M
Rennert
Paul D
PD
Weinreb
Paul H
PH
Ward
Richard
R
Tiao
Greg
G
D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D001652:Bile Ducts / Q000166:cytology* / Q000378:metabolism*; D001656:Biliary Atresia / Q000821:virology*; D002460:Cell Line; D004195:Disease Models, Animal; D020868:Gene Silencing; D022781:Hepatocytes / Q000378:metabolism; D038982:Integrin alpha2beta1 / Q000235:genetics / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D012401:Rotavirus / Q000502:physiology*; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D038982:Integrin alpha2beta1