Ex Vivo T Cell Cytokine Expression Predicts Survival in Patients with Severe Alcoholic Hepatitis

Abstract

Alcoholic hepatitis (AH) is an acute inflammatory liver condition with high early mortality rate. Steroids improve shortterm survival but nonresponders have the worst outcomes. There is a clinical need to identify these high-risk individuals at the time of presentation. T cells are implicated in AH and steroid responsiveness. We measured ex vivo T cell cytokine expression as a candidate biomarker of outcomes in patients with AH. Consecutive patients (bilirubin >80 µmol/L and ratio of aspartate aminotransferase to alanine aminotransferase >1.5 who were heavy alcohol consumers with discriminant function [DF] ≥32), were recruited from University Hospitals Plymouth NHS Trust. T cells were obtained and stimulated ex vivo. Cytokine expression levels were determined by flow cytometry and protein multiplex analysis. Twenty-three patients were recruited (10 male; median age 51 years; baseline DF 67; 30% 90-day mortality). Compared to T cells from nonsurvivors at day 90, T cells from survivors had higher baseline baseline intracellular interleukin (IL)-10:IL-17A ratio (0.43 vs 1.20, p=0.02). Multiplex protein analysis identified interferon γ (IFNγ) and tumor necrosis factor-α (TNF-α) as independent predictors of 90-day mortality (p=0.04, p=0.01, respectively). The ratio of IFNγ to TNF-α was predictive of 90-day mortality (1.4 vs 0.2, p=0.03). These data demonstrate the potential utility of T cell cytokine release assays performed on pretreatment blood samples as biomarkers of survival in patients with severe AH. Our key findings were that intracellular IL-10:IL-17A and IFNγ:TNF-α in culture supernatants were predictors of 90-day mortality. This offers the promise of developing T cell-based diagnostic tools for risk stratification.

Assessment of immune function and prediction of survival and infection in patients with severe alcoholic hepatitis: An exploratory study

“…However, these studies did not examine the relationship between IL‐10 and infection in AH patients. In contrast to the current study, a T‐cell cytokine release assay predicted survival in AH patients, with increased production of anti‐inflammatory interleukin (IL)‐10 associated with death 24 . Another study found that polymorphisms in genes resulting in high TNF‐α and low IL‐10 production were associated with higher rates of sepsis in patients with AH 25 .…”

“…Levels of IFN‐γ were found to be reduced in the serum of patients with chronic ARLD 30 . In a cytokine release assay, levels of T‐cell production of IFN‐γ was predictive of 90‐day mortality in AH patients 24 . Furthermore, IFN‐γ‐induced genes were significantly decreased in severe AH patients and these patients were highly susceptible to infections 29 .…”

“… 30 In a cytokine release assay, levels of T‐cell production of IFN‐γ was predictive of 90‐day mortality in AH patients. 24 Furthermore, IFN‐γ‐induced genes were significantly decreased in severe AH patients and these patients were highly susceptible to infections. 29 In the present study, IFN‐γ measured by standard QFM was strongly associated with 28‐ and 90‐day mortality.…”

“…In contrast to the current study, a T‐cell cytokine release assay predicted survival in AH patients, with increased production of anti‐inflammatory interleukin (IL)‐10 associated with death. 24 Another study found that polymorphisms in genes resulting in high TNF‐α and low IL‐10 production were associated with higher rates of sepsis in patients with AH. 25 In the context of sepsis, IL‐10 is higher in non‐survivors compared to survivors 26 and polymorphisms associated with high IL‐10 production are associated with higher risk of bacterial sepsis.…”

Oxidative stress in alcohol-related liver disease

“…Soluble inflammatory mediators including interleukins have been implicated in ALD[ 60 , 89 , 90 ] and are associated with outcome in patients with AH[ 91 ]. Elevated serum IL-6 levels have recently been identified as a predictor of mortality in severe AH patients[ 64 ].…”