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Author Notes:

Arash Grakoui, PhD, Division of Infectious diseases, Yerkes National Primate Research Center, Division of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30329, Telephone: (404) 727-5850; Fax: (404) 727-7768; arash.grakoui@emory.edu.

D.T. and M.T. conceived and performed the experiments, interpreted the data and co-wrote the manuscript; C.Y.C and E.M.B performed the bacteriological experiments and analysis; P.S. and E.J.E. performed the experiments with D.T. and M.T.; S.G. analyzed the histological slides for scoring, interpretation and provided valuable suggestions; Y.G., M.G. and J. L. performed the deep sequencing analysis for 16S RNA and provided analysis and interpretation; M.M. provided assistance with deep sequencing analysis and interpretation; W.H.K, J.F.M and A.B.A performed transplant surgeries on patients; D.S.W. provided assistance for bacterial identification and analysis, conceived and designed experiments, interpreted the data and provided critical scientific and intellectual input.

A.G. conceived and designed the experiments, interpreted the data and co-wrote the manuscript, with all authors contributing to writing and providing feedback.

We are grateful to the patient donors, Emory Transplant Center team: surgical staff, nursing staff, and coordinators, specifically Sallie Carpentier, Farzan Saeed, Shine Thomas, Ada Ghali, Rivka Elbein and Elizabeth Ferry for their assistance and cooperation throughout this study.

We thank Dr. Pablo Pereira, Pasteur Institute, France for kindly providing us anti-mouse TCRVγ4 (49.2) and TCRVγ7 (F2.64) antibodies.

We are also grateful to Flow Cytometry;Immunology and Pathology Cores of Emory Vaccine Center; and veterinary and animal care staff of Yerkes National Primate Research Center, Emory University for their assistance.

The authors have no conflicts of interest to disclose.


Research Funding:

This project was supported by NIH grants R01AI070101, R01AI124680, R01AI126890, and R01AI136533 to A.G.; and ORIP/OD P51OD011132 (formerly NCRR P51RR000165) to the Yerkes National Primate Research Center (AG).

M.T. is supported by National Institute of Diabetes, and Digestive and Kidney Diseases (NIDDK) Mentored Career Development Award (K01DK109025).

This work was facilitated by the Immunology and Flow Cytometry Core of the Center for AIDS Research at Emory University (P30AI050409).


  • Science & Technology
  • Life Sciences & Biomedicine
  • Gastroenterology & Hepatology
  • PSC
  • Intestinal Microbiota
  • Liver Disease Pathogenesis
  • Immune Response
  • MICE

Alterations in Intestinal Microbiota Lead to Production of Interleukin 17 by Intrahepatic gamma delta T-Cell Receptor-Positive Cells and Pathogenesis of Cholestatic Liver Disease

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Journal Title:



Volume 154, Number 8


, Pages 2178-2193

Type of Work:

Article | Post-print: After Peer Review


Background & Aims: Variants at the ABCB4 or MDR2 locus, which encodes a biliary transport protein, are associated with a spectrum of cholestatic liver diseases. Exacerbation of liver disease has been linked to increased hepatic levels of interleukin (IL) 17, yet the mechanisms of this increase are not understood. We studied mice with disruption of Mdr2 to determine how defects in liver and alteration in the microbiota contribute to production of IL17 by intrahepatic γδ T cells. Methods: We performed studies with Mdr2-/- and littermate FVB/NJ (control) mice. IL17 was measured in serum samples by an enzyme-linked immunosorbent assay. Mice were injected with neutralizing antibodies against the γδ T-cell receptor (TCR; anti-γδ TCR) or mouse IL17A (anti-IL17A). Livers were collected and bacteria were identified in homogenates by culture procedures; TCRγδ+ cells were isolated by flow cytometry. Fecal samples were collected from mice and analyzed by 16S ribosomal DNA sequencing. Cells were stimulated with antibodies or bacteria, and cytokine production was measured. We obtained tissues from 10 patients undergoing liver transplantation for primary sclerosing cholangitis or chronic hepatitis C virus infection. Tissues were analyzed for cytokine production by γδ TCR+ cells. Results: Mdr2–/– mice had collagen deposition around hepatic bile ducts and periportal–bridging fibrosis with influx of inflammatory cells and increased serum levels of IL17 compared with control mice. Administration of anti-IL17A reduced hepatic fibrosis. Livers from Mdr2–/– mice had increased numbers of IL17A+ γδTCR+ cells—particularly of IL17A+ Vγ6Jγ1 γδ TCR+ cells. Fecal samples from Mdr2–/– mice were enriched in Lactobacillus, and liver tissues were enriched in Lactobacillus gasseri compared with control mice. Mdr2–/– mice also had increased intestinal permeability. The γδ TCR+ cells isolated from Mdr2-/- livers produced IL17 in response to heat-killed L gasseri. Intraperitoneal injection of control mice with L gasseri led to increased serum levels of IL17 and liver infiltration by inflammatory cells; injection of these mice with anti-γδ TCR reduced serum level of IL17. Intravenous injections of Mdr2-/- mice with anti-γδ TCR reduced fibrosis; liver levels of IL17, and inflammatory cells; and serum levels of IL17. γδTCR+ cells isolated from livers of patients with primary sclerosing cholangitis, but not hepatitis C virus infection, produced IL17. Conclusions: In Mdr2-/- mice, we found development of liver fibrosis and inflammation to require hepatic activation of γδ TCR+ cells and production of IL17 mediated by exposure to L gasseri. This pathway appears to contribute to development of cholestatic liver disease in patients.

Copyright information:

© 2018 AGA Institute

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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