title: Combination of Mass Cytometry and Imaging Analysis Reveals Origin, Location, and Functional Repopulation of Liver Myeloid Cells in Mice.
abstract: Resident macrophages are derived from yolk sac precursors and seed the liver during embryogenesis. Native cells may be replaced by bone marrow precursors during extensive injuries, irradiation, and infections. We investigated the liver populations of myeloid immune cells and their location, as well as the dynamics of phagocyte repopulation after full depletion. The effects on liver function due to the substitution of original phagocytes by bone marrow-derived surrogates were also examined.
  
  
  
  We collected and analyzed liver tissues from C57BL/6 (control), LysM-EGFP, B6 ACTb-EGFP, CCR2-/-, CD11c-EYFP, CD11c-EYFP-DTR, germ-free mice, CX3CR1gfp/gfp, CX3CR1gpf/wt, and CX3CR1-DTR-EYFP. Liver nonparenchymal cells were immunophenotyped using mass cytometry and gene expression analyses. Kupffer and dendritic cells were depleted from mice by administration of clodronate, and their location and phenotype were examined using intravital microscopy and time-of-flight mass cytometry. Mice were given acetaminophen gavage or intravenous injections of fluorescently labeled Escherichia coli, blood samples were collected and analyzed, and liver function was evaluated. We assessed cytokine profiles of liver tissues using a multiplexed array.
  
  
  
  Using mass cytometry and gene expression analyses, we identified 2 populations of hepatic macrophages and 2 populations of monocytes. We also identified 4 populations of dendritic cells and 1 population of basophils. After selective depletion of liver phagocytes, intravascular myeloid precursors began to differentiate into macrophages and dendritic cells; dendritic cells migrated out of sinusoids, after a delay, via the chemokine CX3CL1. The cell distribution returned to normal in 2 weeks, but the repopulated livers were unable to fully respond to drug-induced injury or clear bacteria for at least 1 month. This defect was associated with increased levels of inflammatory cytokines, and dexamethasone accelerated the repopulation of liver phagocytes.
  
  
  
  In studies of hepatic phagocyte depletion in mice, we found that myeloid precursors can differentiate into liver macrophages and dendritic cells, which each localize to distinct tissue compartments. During replenishment, macrophages acquire the ability to respond appropriately to hepatic injury and to remove bacteria from the blood stream.
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Bruna Araujo
authors: BA
authors: David
authors: Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.
authors: Rafael Machado
authors: RM
authors: Rezende
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Maísa Mota
authors: MM
authors: Antunes
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Mônica Morais
authors: MM
authors: Santos
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Maria Alice
authors: MA
authors: Freitas Lopes
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Ariane Barros
authors: AB
authors: Diniz
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Rafaela Vaz
authors: RV
authors: Sousa Pereira
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Sarah Cozzer
authors: SC
authors: Marchesi
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Débora Moreira
authors: DM
authors: Alvarenga
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Brenda Naemi
authors: BN
authors: Nakagaki
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Alan Moreira
authors: AM
authors: Araújo
authors: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Daniela Silva
authors: DS
authors: Dos Reis
authors: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Renata Monti
authors: RM
authors: Rocha
authors: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Pedro Elias
authors: PE
authors: Marques
authors: University of Calgary, Alberta, Canada.
authors: Woo-Yong
authors: WY
authors: Lee
authors: University of Calgary, Alberta, Canada.
authors: Justin
authors: J
authors: Deniset
authors: University of Calgary, Alberta, Canada.
authors: Pei Xiong
authors: PX
authors: Liew
authors: Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.
authors: Stephen
authors: S
authors: Rubino
authors: Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.
authors: Laura
authors: L
authors: Cox
authors: Resolution of Inflammation Lab, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Vanessa
authors: V
authors: Pinho
authors: Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, São Paulo, Brazil.
authors: Thiago Mattar
authors: TM
authors: Cunha
authors: Centro de Pesquisas René Rachou, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil.
authors: Gabriel Rocha
authors: GR
authors: Fernandes
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil; Departamento de Fisiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: André Gustavo
authors: AG
authors: Oliveira
authors: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors: Mauro Martins
authors: MM
authors: Teixeira
authors: University of Calgary, Alberta, Canada. Electronic address: pkubes@ucalgary.ca.
authors: Paul
authors: P
authors: Kubes
authors: Center for Gastrointestinal Biology, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil. Electronic address: menezesgb@ufmg.br.
authors: Gustavo Batista
authors: GB
authors: Menezes
mesh_terms: D000082:Acetaminophen; D000818:Animals; D015703:Antigens, CD / Q000032:analysis*; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D002454:Cell Differentiation; D056486:Chemical and Drug Induced Liver Injury / Q000276:immunology; D054428:Chemokine CX3CL1 / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D003713:Dendritic Cells / Q000737:chemistry; D016130:Immunophenotyping / Q000379:methods; D000069416:Intravital Microscopy; D037102:Lectins / Q000235:genetics; D008099:Liver / Q000166:cytology* / Q000276:immunology / Q000378:metabolism / Q000503:physiopathology*; D008264:Macrophages / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D055806:Microvessels / Q000378:metabolism; D009000:Monocytes / Q000737:chemistry; D022423:Myeloid Cells / Q000737:chemistry / Q000502:physiology*; D010641:Phenotype; D059467:Transcriptome
chemical_list: D015703:Antigens, CD; D054428:Chemokine CX3CL1; D016207:Cytokines; D037102:Lectins; D000082:Acetaminophen
keywords: CyTOF; DC; Liver Development; Mouse Model