TBA (16S139)

In vitro systems to study natural resistance to HCV-infection

Author(s)

In vitro systems to study natural resistance to HCV-infection

Department(s)/Institutions

Lena Fischer, Ann Byrne, Mark W. Robinson, Cliona O'Farrelly

Introduction

In 1977-79 batches of anti-D immunoglobulin used to prevent Rhesus isoimmunisation in pregnant women were contaminated with HCV from a single donor. Interestingly, up to 40% of the women who received HCV-contaminated anti-D show no evidence of infection. We presume that these women are naturally resistant to HCV due to a highly effective innate immune response that allowed them to clear the virus without mounting an adaptive immune response. These women are potentially of considerable value in the examination of mechanisms that define resistance to HCV-infection.

Aims/Background

So far, the study of innate immune mechanisms in the liver relied on primary hepatocytes which are difficult to obtain and show a restricted life-time. Both multi- and pluripotent cells (MSCs and iPSCs, respectively) are powerful tools to generate patient-specific models to examine host-pathogen interactions. Differentiation of iPSCs into hepatocytes from women who were exposed to HCV during anti-D immunisation (natural resistance, cleared and chronic infection) allows the study of innate host responses in the liver.

Method

MSCs were previously detected in liver perfusates in our lab and were characterised by flow cytometry and their potential to differentiate into osteocytes and adiposites. To generate MSC-derived hepatocytes, cells were successively exposed to different growth factors that mimic liver development. Morphological changes were observed and gene expression profiles of MSC pre and post differentiation were assessed by qPCR. Expression of both pluripotency (sox2, klf4) and hepatocyte (albumin, ck18) markers was determined and gene expression of liver biopsies used as control.

Results

Isolated MSCs from liver perfusate were CD90+/CD105+/CD166+ and CD45-/CD34- and could differentiate into adiposites and osteocytes. Gene expression profiles of MSCs showed upregulation of stem cell markers and absence of liver markers. Hepatocyte differentiation will be further optimised.

Conclusions

In vitro systems that use iPSC-derived hepatocytes will allow the examination of patient-specific immune responses while circumventing the need for primary cells