title: Ectopic expression of murine CD47 minimizes macrophage rejection of human hepatocyte xenografts in immunodeficient mice.
abstract: Macrophages play an important role in the rejection of xenogeneic cells and therefore represent a major obstacle to generating chimeric mice with human xenografts that are useful tools for basic and preclinical medical research. The signal inhibitory regulatory protein α (SIRPα) receptor is a negative regulator of macrophage phagocytic activity and interacts in a species-specific fashion with its ligand CD47. Furthermore, SIRPα polymorphism in laboratory mouse strains significantly affects the extent of human CD47-mediated toleration of human xenotransplants. Aiming to minimize macrophage activity and thus optimize human cell engraftment in immunodeficient mice, we lentivirally transduced murine CD47 (Cd47) into human liver cells. Human HepG2 liver cells expressing Cd47 were less frequently contacted and phagocytosed by murine RAW264.7 macrophages in vitro than their Cd47-negative counterparts. For the generation of human-mouse chimeric livers in immunodeficient BALB-ΔRAG/γ(c) -uPA (urokinase-type plasminogen activator) mice, freshly thawed cryopreserved human hepatocytes were transduced with a lentiviral expression vector for Cd47 using a refined in vitro transduction protocol immediately before transplantation. In vivo, Cd47-positive human primary hepatocytes were selectively retained following engraftment in immunodeficient mice, leading to at least a doubling of liver repopulation efficiencies.
  
  
  CONCLUSIONS
  We conclude that ectopic expression of murine Cd47 in human hepatocytes selectively favors engraftment upon transplantation into mice, a finding that should have a profound impact on the generation of robust humanized small animal models. Moreover, dominance of ectopically expressed murine Cd47 over endogenous human CD47 should also widen the spectrum of immunodeficient mouse strains suitable for humanization.
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Johan M
authors.initials: JM
authors.lastname: Waern
authors.forename: Qinggong
authors.initials: Q
authors.lastname: Yuan
authors.forename: Urda
authors.initials: U
authors.lastname: Rüdrich
authors.forename: Pablo D
authors.initials: PD
authors.lastname: Becker
authors.forename: Kai
authors.initials: K
authors.lastname: Schulze
authors.forename: Helene
authors.initials: H
authors.lastname: Strick-Marchand
authors.forename: Nicholas D
authors.initials: ND
authors.lastname: Huntington
authors.forename: Behrend J
authors.initials: BJ
authors.lastname: Zacher
authors.forename: Karsten
authors.initials: K
authors.lastname: Wursthorn
authors.forename: James P
authors.initials: JP
authors.lastname: DiSanto
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Guzman
authors.forename: Michael P
authors.initials: MP
authors.lastname: Manns
authors.forename: Michael
authors.initials: M
authors.lastname: Ott
authors.forename: Michael
authors.initials: M
authors.lastname: Bock
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D051928:CD47 Antigen / Q000235:genetics / Q000276:immunology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006084:Graft Rejection / Q000235:genetics / Q000276:immunology; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D016867:Immunocompromised Host; D008264:Macrophages / Q000276:immunology / Q000637:transplantation; D051379:Mice; D008807:Mice, Inbred BALB C; D023421:Models, Animal; D011897:Random Allocation; D011971:Receptors, Immunologic / Q000235:genetics* / Q000378:metabolism; D012680:Sensitivity and Specificity; D014183:Transplantation, Heterologous / Q000276:immunology
chemical_list: D000943:Antigens, Differentiation; D051928:CD47 Antigen; C501922:Sirpa protein, mouse; D011971:Receptors, Immunologic; C501920:SIRPA protein, human