To permit mechanistic characterisation of the effects of acute alcohol administration on hepatic cells we will combine ex vivo and in vitro approaches. This will allow us to link biomarker profiles with immune functionality, and test the consequences of hepatic exposure to bacterial derived products such as pathogen associated molecular pattern molecules (PAMPs).
Our methods also permit identification of novel immunomodulatory targets and mechanistic validation of existing targets.
Liver biopsy tissue will be used for detailed phenotyping of infiltrating macrophages, lymphocytes, NK and T cells using confocal immunohistochemistry.
Explanted liver will be used for immunophenotyping of hepatic immune cell populations by cytometry.
We will use non-parenchymal cells collected from these livers to perform flow-based adhesion assays to assess contribution of key target molecules such as IL-1b, taurocholate and ASK1 inhibitor to immune cell recruitment and parenchymal cell function.
We will also use organ culture systems to assess the impact of alcohol and bacterial products on immune and parenchymal cell activation and survival, and consequences of cytokine administration on these responses.
Histological specimens will be used to analyse the expression of proinflammatory chemokines and cytokines and in parallel FACS analysis will be used to characterise the circulating monocyte and lymphocyte populations over the disease course.