Human Hepatocytes and Differentiated Adult-Derived Human Liver Stem/Progenitor Cells Display In Vitro Immunosuppressive Properties Mediated, at Least in Part, through the Nonclassical HLA Class I Molecule HLA-G

One of the main challenges in liver cell therapy (LCT) is the induction of a tolerogenic microenvironment to promote graft acceptance in the recipient. Little is known about the immunomodulatory potential of the hepatic cells used in liver cell therapy. In this work, we wanted to evaluate the immunosuppressive properties of human hepatocytes and adult-derived human liver stem/progenitor cells (ADHLSCs), as well as the potential involvement of the immunomodulatory molecule HLA-G. We demonstrated that both cell types were capable of inhibiting the proliferative response of PBMCs to an allogenic stimulus and that the immune inhibitory potential of ADHLSCs, although lower than that of hepatocytes, increased after hepatogenic differentiation. We demonstrated that liver cells express HLA-G and that the immune inhibition pattern was clearly associated to its expression. Interestingly, HLA-G expression increased after the third step of differentiation, wherein oncostatin M (OSM) was added. A 48 hr treatment with OSM was sufficient to induce HLA-G expression in ADHLSCs and result in immune inhibition. Surprisingly, blocking HLA-G partially reversed the immune inhibition mediated by hepatocytes and differentiated ADHLSCs, but not that of undifferentiated ADHLSCs, suggesting that additional immune inhibitory mechanisms may be used by these cells. In conclusion, we demonstrated that both hepatocytes and ADHLSCs present immunomodulatory properties mediated, at least in part, through HLA-G, which can be upregulated following hepatogenic differentiation or liver cell pretreatment with OSM. These observations open up new perspectives for the induction of tolerance following LCT and for potential therapeutic applications of these liver cells.

Download Full-text

Immunoprofiling of Adult-Derived Human Liver Stem/Progenitor Cells: Impact of Hepatogenic Differentiation and Inflammation

Stem Cells International ◽

10.1155/2017/2679518 ◽

2017 ◽

Vol 2017 ◽

pp. 1-15 ◽

Cited By ~ 9

Author(s):

Hoda El-Kehdy ◽

Camillo Sargiacomo ◽

Mohammad Fayyad-Kazan ◽

Hussein Fayyad-Kazan ◽

Catherine Lombard ◽

...

Keyword(s):

Progenitor Cells ◽

Human Liver ◽

Cell Populations ◽

Optimal Conditions ◽

Immunological Profile ◽

Hepatogenic Differentiation ◽

Key Factor ◽

The Impact ◽

Immune Profiling

Adult-derived human liver stem/progenitor cells (ADHLSCs) are, nowadays, developed as therapeutic medicinal product for the treatment of liver defects. In this study, the impact of hepatogenic differentiation and inflammation priming on the ADHLSCs’ immune profile was assessed in vitro and compared to that of mature hepatocytes. The constitutive immunological profile of ADHLSCs was greatly different from that of hepatocytes. Differences in the expression of the stromal markers CD90 and CD105, adhesion molecules CD44 and CD49e, immunoregulatory molecules CD73 and HO-1, and NK ligands CD112 and CD155 were noted. While they globally preserved their immunological profile in comparison to undifferentiated counterparts, differentiated ADHLSCs showed a significant downregulation of CD200 expression as in hepatocytes. This was mainly induced by signals issued from EGF and OSM. On the other hand, the impact of inflammation was quite similar for all studied cell populations with an increased expression level of CD54 and CD106 and induction of that of CD40 and CD274. In conclusion, our immune profiling study suggests CD200 as a key factor in regulating the immunobiology of differentiated ADHLSCs. A better understanding of the molecular and physiological events related to such marker could help in designing the optimal conditions for an efficient therapeutic use of ADHLSCs.

Download Full-text

Human liver cell cultivation for long-term in vitro toxicity studies in a miniaturized multi-compartment 3D bioreactor system

Zeitschrift für Gastroenterologie ◽

10.1055/s-0030-1269553 ◽

2011 ◽

Vol 49 (01) ◽

Author(s):

SA Hoffmann ◽

M Lübberstedt ◽

U Müller-Vieira ◽

D Knobeloch ◽

A Nüssler ◽

...

Keyword(s):

Liver Cell ◽

Human Liver ◽

In Vitro Toxicity ◽

Cell Cultivation ◽

Toxicity Studies ◽

Human Liver Cell ◽

Bioreactor System

Download Full-text

AAnti-Inflammatory Effects of Plasma Circulating Exosomes Obtained from Normal-Weight and Obese Subjects on Hepatocytes

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530320666200505121426 ◽

2020 ◽

Vol 20 ◽

Author(s):

Reza Afrisham ◽

Sahar Sadegh-Nejadi ◽

Reza Meshkani ◽

Solaleh Emamgholipour ◽

Molood Bagherieh ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Hepg2 Cells ◽

Human Liver ◽

Liver Cells ◽

Normal Weight ◽

Protein Levels ◽

Human Liver Cells ◽

Tnf Α ◽

Anti Inflammatory

Introduction: Obesity is a disorder with low-grade chronic inflammation that plays a key role in the hepatic inflammation and steatosis. Moreover, there are studies to support the role of exosomes in the cellular communications, the regulation of metabolic homeostasis and immunomodulatory activity. Accordingly, we aimed to evaluate the influence of plasma circulating exosomes derived from females with normal-weight and obesity on the secretion of inflammatory cytokines in human liver cells. Methods: Plasma circulating exosomes were isolated from four normal (N-Exo) and four obese (O-Exo) women. The exosomes were characterized and approved for CD63 expression (common exosomal protein marker) and morphology/size using the western blot and TEM methods, respectively. The exosomes were used for stimulation of HepG2 cells in vitro. After 24 h incubation, the protein levels of TNF-α,IL-6, and IL-1β were measured in the culture supernatant of HepG2 cells using the ELISA kit. Results: The protein levels of IL-6 and TNF-α in the cells treated with O-Exo and N-Exo reduced significantly in comparison with control group (P=0.039 and P<0.001 respectively), while significance differences were not found between normal and obese groups (P=0.808, and P=0.978 respectively). However, no significant differences were found between three groups in term of IL-1β levels (P=0.069). Based on the correlation analysis, the protein levels of IL-6 were positively correlated with TNF-α (r 0.978, P<0.001). Conclusion: These findings suggest that plasma circulating exosomes have probably anti-inflammatory properties independently from body mass index and may decrease the secretion of inflammatory cytokines in liver. However, further investigations in vitro and in vivo are needed to address the anti-inflammatory function of N-Exo and O-Exo in human liver cells and/or other cells.

Download Full-text

The effect of liver donors’ age, gender and metabolic state on pancreatic lineage activation

Regenerative Medicine ◽

10.2217/rme-2020-0092 ◽

2021 ◽

Vol 16 (1) ◽

pp. 19-31

Author(s):

Ioan V Matei ◽

Irit Meivar-Levy ◽

Daniela Lixandru ◽

Simona Dima ◽

Ioana R Florea ◽

...

Keyword(s):

Replacement Therapy ◽

Human Liver ◽

Liver Cells ◽

Diabetic Patients ◽

Metabolic State ◽

Human Liver Cells ◽

Metabolic States ◽

Different Ages ◽

Liver Donors

Autologous cells replacement therapy by liver to pancreas transdifferentiation (TD) allows diabetic patients to be also the donors of their own therapeutic tissue. Aim: To analyze whether the efficiency of the process is affected by liver donors’ heterogeneity with regard to age, gender and the metabolic state. Materials & methods: TD of liver cells derived from nondiabetic and diabetic donors at different ages was characterized at molecular and cellular levels, in vitro. Results: Neither liver cells proliferation nor the propagated cells TD efficiency directly correlate with the age (3–60 years), gender or the metabolic state of the donors. Conclusion: Human liver cells derived from a wide array of ages and metabolic states can be used for autologous cells therapies for diabetics.

Download Full-text

Identification of Human Sulfotransferases Active towards Silymarin Flavonolignans and Taxifolin

Metabolites ◽

10.3390/metabo10080329 ◽

2020 ◽

Vol 10 (8) ◽

pp. 329

Author(s):

Jiří Vrba ◽

Barbora Papoušková ◽

Pavel Kosina ◽

Kateřina Lněničková ◽

Kateřina Valentová ◽

...

Keyword(s):

Human Liver ◽

High Performance ◽

Herbal Remedy ◽

Expression Patterns ◽

Human Hepatocytes ◽

Tandem Mass ◽

Recombinant Enzymes ◽

Metabolic Reactions ◽

Natural Phenolic Compounds

Natural phenolic compounds are known to be metabolized by phase II metabolic reactions. In this study, we examined the in vitro sulfation of the main constituents of silymarin, an herbal remedy produced from the fruits of the milk thistle. The study focused on major flavonolignan constituents, including silybin A, silybin B, isosilybin A, isosilybin B, silychristin, and silydianin, as well as the flavonoid taxifolin. Using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS), individual flavonolignans and taxifolin were found to be sulfated by human liver and human intestinal cytosols. Moreover, experiments with recombinant enzymes revealed that human sulfotransferases (SULTs) 1A1*1, 1A1*2, 1A2, 1A3, 1B1, 1C4, and 1E1 catalyzed the sulfation of all of the tested compounds, with the exception of silydianin, which was not sulfated by SULT1B1 and SULT1C4. The sulfation products detected were monosulfates, of which some of the major ones were identified as silybin A 20-O-sulfate, silybin B 20-O-sulfate, and isosilybin A 20-O-sulfate. Further, we also observed the sulfation of the tested compounds when they were tested in the silymarin mixture. Sulfates of flavonolignans and of taxifolin were produced by incubating silymarin with all of the above SULT enzymes, with human liver and intestinal cytosols, and also with human hepatocytes, even though the spectrum and amount of the sulfates varied among the metabolic models. Considering our results and the expression patterns of human sulfotransferases in metabolic tissues, we conclude that flavonolignans and taxifolin can potentially undergo both intestinal and hepatic sulfation, and that SULTs 1A1, 1A3, 1B1, and 1E1 could be involved in the biotransformation of the constituents of silymarin.

Download Full-text