Alloresponse Is Associated With Major
Histocompatibility Complex Class II
Up-Regulation on Hepatocytes and
Suppressible by Regulatory T Cells
Daphne E. DeTemple,
Christine S. Falk,
and Florian W. R. Vondran
Regenerative Medicine and Experimental Surgery, Department of General, Visceral and Transplant Surgery,
Transplant Immunology, Integrated Research and Treatment Centre Transplantation,
Institute for Transfusion Medicine, and
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany; and
Centre for Infection Research, partner site Hannover-Braunschweig, Hannover, Germany
Hepatocyte transplantation is a promising therapeutic approach for various liver diseases. Despite the liver’s tolerogenic potential,
early immune-mediated loss of transplanted cells is observed, and longterm acceptance has not been achieved yet. Patients deemed
tolerant after liver transplantation presented an increased frequency of regulatory T cells (Tregs), which therefore also might enable
reduction of posttransplant cell loss and enhance longterm allograft acceptance. We hence characterized hepatocyte-induced
immune reactions and evaluated the immunomodulatory potential of Tregs applying mixed lymphocyte cultures and mixed lym-
phocyte hepatocyte cultures. These were set up using peripheral blood mononuclear cells and primary human hepatocytes, respec-
tively. Polyclonally expanded CD4
Tregs were added to cocultures in single-/trans-well setups with/without
supplementation of anti-interferon c (IFNc) antibodies. Hepatocyte-induced alloresponses were then analyzed by multicolor ﬂow
cytometry. Measurements indicated that T cell response upon stimulation was associated with IFNc-induced major histocompati-
bility complex (MHC) class II up-regulation on hepatocytes and mediated by CD4
T cells. An indirect route of antigen presenta-
tion could be ruled out by use of fragmented hepatocytes and culture supernatants of hepatocytes. Allospeciﬁc proliferation was
accompanied by inﬂammatory cytokine secretion. CD8
T cells showed early up-regulation of CD69 despite lack of cell prolifera-
tion in the course of coculture. Supplementation of Tregs effectively abrogated hepatocyte-induced alloresponses and was primarily
cell contact dependent. In conclusion, human hepatocytes induce a CD4
T cell alloresponse in vitro, which is associated with
MHC class II up-regulation on hepatocytes and is susceptible to suppression by Tregs.
Liver Transplantation 24 407–419 2018 AASLD.
Received July 2, 2017; accepted December 16, 2017.
Hepatocyte transplantation (HT) is a promising thera-
peutic approach as treatment for various liver diseases.
Primary human hepatocytes (PHHs) may be cryopre-
served for use of HT in emergencies
modiﬁed extracorporally prior to transplantation.
animal experiments, HT leads to hepatic remodeling
with histologically indistinguishable engrafted hepato-
These achievements could not yet be transferred
into clinical practice, where HT only resulted in tran-
sient amelioration of liver function
for up to 52 days, before patients require orthotopic liver
Reasons for the limited cell survival
might be competition with tissue-resident cells in a
and rejection by the
recipient’s immune system.
Rare occurrence of hyperacute rejection and immu-
nomodulating effects in combined hepatorenal
Abbreviations: FasL, Fas Ligand; FOXP3, forkhead box P3; HLA,
human leukocyte antigen; HT, hepatocyte transplantation; IFNc,
interferon c; IL, interleukin; MFI, mean ﬂuorescence intensity;
MHC, major histocompatibility complex; MLC, mixed lymphocyte
culture; MLHC, mixed lymphocyte hepatocyte culture; PBMC,
peripheral blood mononuclear cell; PHH, primary human hepatocyte;
SEM, standard error of the mean; sCD40L, soluble CD40 ligand;
DETEMPLE ET AL.