Ex vivo T cell cytokine expression predicts survival in patients with severe alcoholic hepatitis

Aim Alcoholic hepatitis (AH) is an acute inflammatory liver condition with high early mortality. Steroids have proven short-term survival benefit but non-responders have the worst outcomes. There is a clinical need to identify these high-risk individuals at presentation. T cells have been implicated in alcoholic hepatitis and steroid responsiveness. We aimed to measure ex vivo T cell cytokine expression as a candidate biomarker of outcome in patients with AH. Methods Consecutive patients with AH (bilirubin >80µmol/L and AST:ALT>1.5 in heavy alcohol consumers with discriminant function [DF]≥32), were recruited from University Hospitals Plymouth NHS Trust. T cells were obtained and stimulated ex vivo before cytokine expression was determined by flow cytometry and protein multiplex analysis. Results Twenty-three patients were recruited (10 male; median age 51; baseline DF 67; 30% 90-day mortality). Compared to non-survivors at day 90, T cells from survivors had higher baseline intracellular IL-10:IL-17A ratio (0.43 v 1.20; p=0.02). Multiplex protein analysis identified IFNγ and TNFα as independent predictors of 90-day mortality (p=0.04 and p=0.01 respectively). The ratio of IFNγ/TNFα was predictive of 90-day mortality (1.4 v 0.2; p=0.03). Conclusions These data demonstrate the potential utility of T cell cytokine release assays performed on pre-treatment blood samples as biomarkers of survival in severe AH. Our key findings were that both the ratio of intracellular IL-10 to IL-17A and the ratio of IFNγ to TNFα in culture supernatants were predictors of 90-day mortality. This offers the promise of developing T cell based diagnostic tools for risk stratification.