title: CD8+ T lymphocyte response against extrahepatic biliary epithelium is activated by epitopes within NSP4 in experimental biliary atresia.
abstract: Interferon (IFN)-γ-driven and CD8+ T cell-dependent inflammatory injury to extrahepatic biliary epithelium (EHBE) is likely to be involved in the development of biliary atresia (BA). We previously showed that viral protein NSP4 is the pathogenic immunogen that causes biliary injury in BA. In this study, NSP4 or four synthetic NSP4 (NSP4(157-170), NSP4(144-152), NSP4(93-110), NSP4(24-32)) identified by computer analysis as candidate CD8+ T cell epitopes were injected into neonatal mice. The pathogenic NSP4 epitopes were confirmed by studying extrahepatic bile duct injury, IFN-γ release and CD8+ T cell response against EHBE. The results revealed, at 7 days postinjection, inoculation of glutathione S-transferase (GST)-NSP4 caused EHBE injury and BA in neonatal mice. At 7 or 14 days postinject, inoculation of GST-NSP4, NSP4(144-152), or NSP4(157-170) increased IFN-γ release by CD8+ T cells, elevated the population of hepatic memory CD8+ T cells, and augmented cytotoxicity of CD8+ T cells to rhesus rotavirus (RRV)-infected or naive EHBE cells. Furthermore, depletion of CD8+ T cells in mice abrogated the elevation of GST-NSP4-induced serum IFN-γ. Lastly, parenteral immunization of mouse dams with GST-NSP4, NSP4(144-152), or NSP4(157-170) decreased the incidence of RRV-induced BA in their offspring. Overall, this study reports the CD8+ T cell response against EHBE is activated by epitopes within rotavirus NSP4 in experimental BA. Neonatal passive immunization by maternal vaccination against NSP4(144-152) or NSP4(157-170) is effective in protecting neonates from developing RRV-related BA.
authors.affiliation: Department of Pediatric Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China; Department of Pediatric Surgery, First Affiliated Hospital of Henan University of Science and Technology, Luoyang, People's Republic of China; and.
authors.forename: Shuaiyu
authors.initials: S
authors.lastname: Zheng
authors.affiliation: Department of Pediatric Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China;
authors.forename: Hongyi
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Pediatric Surgery, First Affiliated Hospital of Henan University of Science and Technology, Luoyang, People's Republic of China; and.
authors.forename: Xiaojin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Pediatric Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China;
authors.forename: Fei
authors.initials: F
authors.lastname: Peng
authors.affiliation: Department of Pediatric Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China;
authors.forename: Xuyong
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Pediatric Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China;
authors.forename: Jixin
authors.initials: J
authors.lastname: Yang
authors.affiliation: Center for Cell and Developmental Biology, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: David
authors.initials: D
authors.lastname: Brigstock
authors.affiliation: Department of Pediatric Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China; fengjiexiong@126.com.
authors.forename: Jiexiong
authors.initials: J
authors.lastname: Feng
mesh_terms: D000818:Animals; D017734:Bile Ducts, Extrahepatic / Q000276:immunology* / Q000473:pathology / Q000821:virology; D001656:Biliary Atresia / Q000276:immunology* / Q000473:pathology / Q000517:prevention & control / Q000821:virology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D004195:Disease Models, Animal; D018984:Epitopes, T-Lymphocyte; D005260:Female; D006023:Glycoproteins / Q000008:administration & dosage / Q000276:immunology*; D007116:Immunization, Passive; D007156:Immunologic Memory; D007274:Injections, Intraperitoneal; D007371:Interferon-gamma / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010446:Peptide Fragments / Q000008:administration & dosage / Q000276:immunology*; D011247:Pregnancy; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000276:immunology; D012400:Rotavirus Infections / Q000276:immunology* / Q000473:pathology / Q000517:prevention & control / Q000821:virology; D022243:Rotavirus Vaccines / Q000008:administration & dosage / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014118:Toxins, Biological / Q000008:administration & dosage / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D022243:Rotavirus Vaccines; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins; C052014:rhesus rotavirus vaccine; D007371:Interferon-gamma
keywords: biliary atresia; epitope mapping; inflammation; pathogenic epitopes; rhesus rotavirus