Antibiotic pretreatment alleviates liver transplant damage in mice and humans
Kojiro Nakamura, … , Ronald W. Busuttil, Jerzy W. Kupiec-Weglinski
Kojiro Nakamura, … , Ronald W. Busuttil, Jerzy W. Kupiec-Weglinski
Published July 22, 2019
Citation Information: J Clin Invest. 2019;129(8):3420-3434. https://doi.org/10.1172/JCI127550.
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Research Article Immunology Transplantation

Antibiotic pretreatment alleviates liver transplant damage in mice and humans

Abstract

Although modifications of gut microbiota with antibiotics (Abx) influence mouse skin and cardiac allografts, its role in orthotopic liver transplantation (OLT) remains unknown. We aimed to determine whether and how recipient Abx pretreatment may affect hepatic ischemia-reperfusion injury (IRI) and OLT outcomes. Mice (C57BL/6) with or without Abx treatment (10 days) were transplanted with allogeneic (BALB/c) cold-stored (18 hours) livers, followed by liver and blood sampling (6 hours). We divided 264 human OLT recipients on the basis of duration of pre-OLT Abx treatment into control (Abx-free/Abx <10 days; n = 108) and Abx treatment (Abx ≥10days; n = 156) groups; OLT biopsy (Bx) samples were collected 2 hours after OLT (n = 52). Abx in mice mitigated IRI-stressed OLT (IRI-OLT), decreased CCAAT/enhancer-binding protein homologous protein (CHOP) (endoplasmic reticulum [ER] stress), enhanced LC3B (autophagy), and inhibited inflammation, whereas it increased serum prostaglandin E2 (PGE2) and hepatic PGE2 receptor 4 (EP4) expression. PGE2 increased EP4, suppressed CHOP, and induced autophagosome formation in hepatocyte cultures in an EP4-dependent manner. An EP4 antagonist restored CHOP, suppressed LC3B, and recreated IRI-OLT. Remarkably, human recipients of Abx treatment plus OLT (Abx-OLT), despite severe pretransplantation clinical acuity, had higher EP4 and LC3B levels but lower CHOP levels, which coincided with improved hepatocellular function (serum aspartate aminotransferase/serum aspartate aminotransferase [sALT/sAST]) and a decreased incidence of early allograft dysfunction (EAD). Multivariate analysis identified “Abx-free/Abx <10 days” as a predictive factor of EAD. This study documents the benefits of Abx pretreatment in liver transplant recipients, identifies ER stress and autophagy regulation by the PGE2/EP4 axis as a homeostatic underpinning, and points to the microbiome as a therapeutic target in OLT.

Authors

Kojiro Nakamura, Shoichi Kageyama, Takahiro Ito, Hirofumi Hirao, Kentaro Kadono, Antony Aziz, Kenneth J. Dery, Matthew J. Everly, Kojiro Taura, Shinji Uemoto, Douglas G. Farmer, Fady M. Kaldas, Ronald W. Busuttil, Jerzy W. Kupiec-Weglinski

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Figure 6

Abx treatment is associated with decreased CHOP and p-S6K, increased LC3B, and lower serum liver enzyme levels (POD1) in OLT recipients.

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Abx treatment is associated with decreased CHOP and p-S6K, increased LC3...
Human OLT Bx samples collected 2 hours after reperfusion (n = 52) were classified into control (recipients with pre-OLT Abx <10 days or without Abx; n = 24) and Abx treatment (recipients with pre-OLT Abx ≥10 days; n= 28) groups (see also Figure 5A). (A) Representative Western blot detection of CHOP, p-S6K, and LC3B (cases 1 and 2: control group; cases 3 and 4: Abx group). (B) Western blot quantification of CHOP, p-S6K, LC3B-I plus LC3B-II, and LC3B-II with β-actin normalization. (C) Representative LC3B and TUNEL staining in human OLT. Arrowheads indicate TUNEL+ dead cells. Original magnification, ×400. (D) qRT-PCR detection of mRNA coding for CD68 and cathepsin G. Data were normalized to GAPDH gene expression. (E) sALT and sAST levels on POD1. Data indicate the mean ± SEM. #P < 0.05, by Mann-Whitney U test.