Silencing of HLA class I on primary human hepatocytes as a novel strategy for reduction in alloreactivity.

Autor: Figueiredo C; Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.; Excellence Cluster REBIRTH - From Regenerative Biology to Reconstructive Therapy, Hannover, Germany., Oldhafer F; ReMediES, Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany., Wittauer EM; ReMediES, Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany., Carvalho-Oliveira M; Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.; Excellence Cluster REBIRTH - From Regenerative Biology to Reconstructive Therapy, Hannover, Germany., Akhdar A; German Centre for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Hannover, Germany., Beetz O; ReMediES, Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany., Chen-Wacker C; Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.; Excellence Cluster REBIRTH - From Regenerative Biology to Reconstructive Therapy, Hannover, Germany., Yuzefovych Y; Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany., Falk CS; German Centre for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Hannover, Germany.; Institute of Transplant Immunology, Hannover Medical School, Hannover, Germany., Blasczyk R; Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany., Vondran FWR; ReMediES, Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany.; German Centre for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Hannover, Germany.
Jazyk: angličtina
Zdroj: Journal of cellular and molecular medicine [J Cell Mol Med] 2019 Aug; Vol. 23 (8), pp. 5705-5714. Date of Electronic Publication: 2019 Jun 10.
DOI: 10.1111/jcmm.14484
Abstrakt: In contrast to the whole liver, primary hepatocytes are highly immunogenic. Thus, alternative strategies of immunomodulation after hepatocyte transplantation are of special interest. Silencing of HLA class I expression is expected to reduce the strength of allogeneic immune responses and to improve graft survival. In this study, primary human hepatocytes (PHH) were isolated using a two-step-collagenase perfusion-technique and co-cultured with allogeneic lymphocytes in terms of a mixed lymphocyte hepatocyte culture. Expression of HLA class I on PHH was silenced using lentiviral vectors encoding for β2-microglobulin-specific short hairpin RNA (shβ2m) or non-specific shRNA (shNS) as control. The delivery of shβ2m into PHH caused a decrease by up to 96% in β2m transcript levels and a down-regulation of HLA class I cell surface expression on PHH by up to 57%. Proliferative T cell alloresponses against HLA-silenced PHH were significantly lower than those observed form fully HLA-expressing PHH. In addition, significantly lower secretion of pro-inflammatory cytokines was observed. Levels of albumin, urea and aspartate-aminotransferase did not differ in supernatants of cultured PHH. In conclusion, silencing HLA class I expression on PHH might represent a promising approach for immunomodulation in the transplant setting without compromising metabolic function of silenced hepatocytes.
(© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)
Databáze: MEDLINE
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