Z Gastroenterol 2014; 52 - P_5_12
DOI: 10.1055/s-0033-1361021

Easy and robust testing of T cell responsiveness to HBV antigens in whole blood of chronic Hepatitis B patients

W Dammermann 1, EM Stiel 1, M Kohring 1, J Schulze zur Wiesch 1, AW Lohse 1, S Lüth 1
  • 1University Medical Center Hamburg-Eppendorf, 1. Department of Medicine, Hamburg, Germany

Background and Aim:

Hyporesponsiveness of HBV-specific T cells in patients with chronic hepatitis B infection has been well documented using advanced and technically demanding analytical methods like flow cytometry or ELISPOT. Interferon gamma release assays (IGRA) have been developed to support the easy and fast diagnosis of diseases like tuberculosis, which lack robust serological test systems. IGRAs focus on cellular immunity especially memory T cells and thus also allow rapid screening prior to complex flow cytometric testing. We hypothesized that an improved whole blood based cytokine release assay is capable of displaying T cell responsiveness to HBV antigens in chronic Hepatitis B patients.

Methods:

28 chronic Hepatitis B patients (CHB), 5 acute hepatitis B patients (AHB) and 11 healthy controls (HC) were tested by ELISA for IFNγ- and IL2-secretion in whole blood after challenge with synthetic peptide libraries of Hepatitis B core antigen (HBcAg) or Hepatitis B surface antigen (HBsAg). Supplementation of whole blood with D-Glucose was used to further enhance cytokine secretion.

Results:

Mean IFNγ response to HBcAg was highest in the AHB group (91 ± 79 pg/ml), whereas CHB patients showed a weaker response (19 ± 24 pg/ml) and HC remained unresponsive (8 ± 12 pg/ml). IL2 levels after HBcAg challenge were also higher in the AHB group compared to CHB and HC (193 ± 240 vs. 8 ± 13 and 2 ± 1 pg/ml, respectively). HBsAg stimulation led to increased IFNγ and IL2 levels in the AHB group and HC (28 ± 35 and 24 ± 17 pg/ml, for AHB; 24 ± 36 and 48 ± 128 pg/ml, for HC). CHB patients developed no IFNγ or IL2 response to HBsAg (9 ± 16 and 2 ± 3 pg/ml, respectively).

Conclusions:

Our improved whole blood based cytokine release assay is able to analyze the hyporesponsiveness of HBV-specific T cells in patients with chronic hepatitis B infection. Thus, it constitutes an easy and robust tool for screening HBV specific cellular immunity in comparison to flow cytometry or ELISPOT.