Comparative Phenotype and Immunogenicity of Freshly Isolated and Immortalized Rat Hepatocytes

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Abstract:

Immortalized hepatocytes are an attractive cell source for hepatocyte transplantation and gene transfer. We compared the phenotype and immunogenicity of freshly isolated (FIH) and immortalized (IMH) rat hepatocytes. Effect of culture and proinflammatory cytokines (TNF-α, IFN-γ) was studied on phenotype. FIH were isolated by collagenase digestion. Two SV40 immortalized hepatocyte cell lines were tested (RH1 and P9). Immunophenotyping was performed by FACS analysis using anti-rat-specific antibodies. Immunogenicity was evaluated by a mixed lymphocyte hepatocyte reaction (MLHR). FIH suspension was an almost homogeneous parenchymal cell population with few (1–2%) CD8+ cells. FIH showed a positive staining for ICAM-1 (20–35%) and for Class I (RT1A, 30–60%) but no staining for Class II (RT1B). After 48 h of culture, the already ICAM-1-positive cells were more strongly stained and additionally 3.6% of the cells (possibly endothelial cells) were Class II positive. IMH showed a consistent expression of Class I (93–97%) and ICAM-1 (95–97%) but no expression of Class II. Culture of IMH for 48 h had no effect on Class II expression but increased ICAM-1 expression. Addition of TNF-α at 1000 UI/ml to cultures of FIH or IMH increased Class I and ICAM-1 expression whereas IFN-γ (50 or 1000 UI/ml) had no evident effect. Hepatocyte immunogenicity, assessed in MLHR and appreciated by the stimulation index (SI) test/SI syngeneic control, was similar for IMH (RH1: 2.68 ± 0.89; P9: 2.37 ± 0.78) and FIH (2.52 ± 0.18). In conclusion, despite some quantitative immunophenotypic differences, FIH and IMH induced the same proliferation rate of allogeneic T lymphocytes. Thus, immortalized hepatocytes may constitute an appropriate cellular model to study the prevention of hepatocyte rejection by gene transfer.

Keywords: Freshly isolated h; Key words: Allotransplantation

Document Type: Research Article

DOI: http://dx.doi.org/10.3727/000000001783986242

Affiliations: 1: †Laboratoire de Thérapeutique Immuno-moléculaire, EFS/EA 2284/Inserm E-0119, 1 Bd A Fleming, BP 1937, 25020 Besançon cedex, France 2: *Laboratoire de Chirurgie Expérimentale, Fondation Transplantation, 5 Avenue Molière, 67200 Strasbourg, France 3: ‡University of Strathclyde, Bioengineering Unit, Wolfson Center, 106 Rottenrow, Glasgow 64 ONW, Scotland 4: §Faculté de Médecine et Pharmacie, Laboratoire de Biologie Cellulaire, Place Saint Jacques, 25030 Besançon, France

Publication date: January 1, 2001

More about this publication?
  • Cell Transplantation publishes original, peer-reviewed research and review articles on the subject of cell transplantation and its application to human diseases. To ensure high-quality contributions from all areas of transplantation, separate section editors and editorial boards have been established. Articles deal with a wide range of topics including physiological, medical, preclinical, tissue engineering, and device-oriented aspects of transplantation of nervous system, endocrine, growth factor-secreting, bone marrow, epithelial, endothelial, and genetically engineered cells, among others. Basic clinical studies and immunological research papers are also featured. To provide complete coverage of this revolutionary field, Cell Transplantation will report on relevant technological advances, and ethical and regulatory considerations of cell transplants. Cell Transplantation is now an Open Access journal starting with volume 18 in 2009, and therefore there will be an inexpensive publication charge, which is dependent on the number of pages, in addition to the charge for color figures. This will allow work to be disseminated to a wider audience and also entitle the corresponding author to a free PDF, as well as prepublication of an unedited version of the manuscript.
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