Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Nov 18, 2016; 8(32): 1370-1383
Published online Nov 18, 2016. doi: 10.4254/wjh.v8.i32.1370
Changes in cellular proliferation and plasma products are associated with liver failure
Juliana Gil Melgaço, Frederico Marianetti Soriani, Pedro Henrique Ferreira Sucupira, Leonardo Assaf Pinheiro, Yasmine Rangel Vieira, Jaqueline Mendes de Oliveira, Lia Laura Lewis-Ximenez, Cristina Carvalho Vianna Araújo, Lúcio Filgueiras Pacheco-Moreira, Gustavo Batista Menezes, Oswaldo Gonçalves Cruz, Claudia Lamarca Vitral, Marcelo Alves Pinto
Juliana Gil Melgaço, Leonardo Assaf Pinheiro, Claudia Lamarca Vitral, Departamento de Microbiologia e Parasitologia, Universidade Federal Fluminense, Niterói 20550-013, Brazil
Juliana Gil Melgaço, Yasmine Rangel Vieira, Jaqueline Mendes de Oliveira, Marcelo Alves Pinto, Instituto Oswaldo Cruz/Fiocruz, Laboratório de Desenvolvimento Tecnológico em Virologia, Rio de Janeiro 21040-900, Brazil
Frederico Marianetti Soriani, Pedro Henrique Ferreira Sucupira, Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil
Lia Laura Lewis-Ximenez, Instituto Oswaldo Cruz/Fiocruz, Ambulatório de Hepatites Virais, Rio de Janeiro 21040-900, Brazil
Cristina Carvalho Vianna Araújo, Lúcio Filgueiras Pacheco-Moreira, Hospital Federal de Bonsucesso, Rio de Janeiro 21041-030, Brazil
Gustavo Batista Menezes, Laboratório de Imunofarmacologia, Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil
Oswaldo Gonçalves Cruz, Fundação Oswaldo Cruz, Programa de Computação Científica, Rio de Janeiro 21040-900, Brazil
Author contributions: All authors have seen and approved the content of the manuscript and contributed meaningfully to the work. In summary, Melgaço JG conceived the study, performed the experiments, analyzed the data and wrote the manuscript; Pinheiro LA, Vieira YR, de Oliveira JM and Vitral CL acquired the data for cytokine quantification, serological data and the virological marker assay; de Oliveira JM reviewed the analysis and wrote the manuscript; Lewis-Ximenez LL collected the clinical data and the samples from the self-limited acute hepatitis patients and healthy subjects; Araújo CCV and Pacheco-Moreira LF collected the clinical data and the samples from the acute liver failure patients; Soriani FM, Sucupira PHF and Menezes GB performed the DNA extraction and quantified the mitochondrial DNA using molecular biology assays; Cruz OG performed the statistical analysis; Vitral CL and Pinto MA participated in the study design and coordination; Pinto MA participated in the analysis of data and the preparation of the manuscript.
Supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq, No. 308951/2010-7; and the Fundação de Amparo à Pesquisas no Rio de Janeiro - Faperj, No. E-26/110.848/2013.
Institutional review board statement: The study protocol was approved by the National Commission on Ethics in Research (CONEP) and by the institutional review board of the Oswaldo Cruz Foundation, FIOCRUZ (222/03).
Conflict-of-interest statement: The authors declare that they have no competing interests.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Correspondence to: Juliana Gil Melgaço, PhD, Instituto Oswaldo Cruz/Fiocruz, Laboratório de Desenvolvimento Tecnológico em Virologia, Fundação Oswaldo Cruz/FIOCRUZ, Avenida Brasil, 4365, pavilhão Hélio e Peggy Pereira, Rio de Janeiro 21040-900, Brazil.
Telephone: +55-21-25621711 Fax: +55-21-25621898
Received: May 23, 2016
Peer-review started: May 25, 2016
First decision: July 20, 2016
Revised: August 3, 2016
Accepted: September 13, 2016
Article in press: September 18, 2016
Published online: November 18, 2016

To study the differences in immune response and cytokine profile between acute liver failure and self-limited acute hepatitis.


Forty-six patients with self-limited acute hepatitis (AH), sixteen patients with acute liver failure (ALF), and twenty-two healthy subjects were involved in this study. The inflammatory and anti-inflammatory products in plasma samples were quantified using commercial enzyme-linked immunoassays and quantitative real-time PCR. The cellular immune responses were measured by proliferation assay using flow cytometry. The groups were divided into viral- and non-viral-induced self-limited AH and ALF. Thus, we worked with five groups: Hepatitis A virus (HAV)-induced self-limited acute hepatitis (HAV-AH), HAV-induced ALF (HAV-ALF), non-viral-induced self-limited acute hepatitis (non-viral AH), non-viral-induced acute liver failure (non-viral ALF), and healthy subjects (HC). Comparisons among HAV and non-viral-induced AH and ALF were performed.


The levels of mitochondrial DNA (mtDNA) and the cytokines investigated [interleukin (IL)-6, IL-8, IL-10, interferon gamma, and tumor necrosis factor] were significantly increased in ALF patients, independently of etiology (P < 0.05). High plasma mtDNA and IL-10 were the best markers associated with ALF [mtDNA: OR = 320.5 (95%CI: 14.42-7123.33), P < 0.0001; and IL-10: OR = 18.8 (95%CI: 1.38-257.94), P = 0.028] and death [mtDNA: OR = 12.1 (95%CI: 2.57-57.07), P = 0.002; and IL-10: OR = 8.01 (95%CI: 1.26-50.97), P = 0.027]. In the cellular proliferation assay, NKbright, NKT and regulatory T cells (TReg) predominated in virus-specific stimulation in HAV-induced ALF patients with an anergic behavior in the cellular response to mitotic stimulation. Therefore, in non-viral-induced ALF, anergic behavior of activated T cells was not observed after mitotic stimulation, as expected and as described by the literature.


mtDNA and IL-10 may be predictors of ALF and death. TReg cells are involved in immunological disturbance in HAV-induced ALF.

Keywords: Acute liver failure, Cytokines, Mitochondrial DNA, Cellular immune response, Hepatitis A virus

Core tip: Acute liver diseases induced by viral infections are considered major causes of liver failure and death in Brazil. To better understand this pathogenesis, we investigated in a pioneering way the cellular immune response, inflammatory mediators and mitochondrial products in patients with hepatitis A virus (HAV)-induced acute liver failure (ALF) in comparison to patients with non-virus-induced ALF in a cross-sectional study. The results showed that non-invasive samples could be helpful to assay early prognostic markers that would indicate the necessity for liver transplantation. The contribution of in vitro immune response involved in ALF can be helpful to show the necessity of mass vaccination against HAV.