Combination of Mass Cytometry and Imaging Analysis Reveals Origin, Location, and Functional Repopulation of Liver Myeloid Cells in Mice

• Published in: Gastroenterology (New York, N.Y. 1943) Vol. 151; no. 6; pp. 1176 - 1191
• Main Authors: David, Bruna Araujo, Rezende, Rafael Machado, Antunes, Maísa Mota, Santos, Mônica Morais, Freitas Lopes, Maria Alice, Diniz, Ariane Barros, Sousa Pereira, Rafaela Vaz, Marchesi, Sarah Cozzer, Alvarenga, Débora Moreira, Nakagaki, Brenda Naemi, Araújo, Alan Moreira, dos Reis, Daniela Silva, Rocha, Renata Monti, Marques, Pedro Elias, Lee, Woo-Yong, Deniset, Justin, Liew, Pei Xiong, Rubino, Stephen, Cox, Laura, Pinho, Vanessa, Cunha, Thiago Mattar, Fernandes, Gabriel Rocha, Oliveira, André Gustavo, Teixeira, Mauro Martins, Kubes, Paul, Menezes, Gustavo Batista
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-12-2016
• Subjects: Acetaminophen; Animals; Antigens, CD - analysis; Bone Marrow Cells - cytology; Bone Marrow Cells - physiology; Cell Differentiation; Chemical and Drug Induced Liver Injury - immunology; Chemokine CX3CL1 - metabolism; CyTOF; Cytokines - genetics; Cytokines - metabolism; Dendritic Cells - chemistry; Gastroenterology and Hepatology; Immunophenotyping - methods; Intravital Microscopy; Lectins - genetics; Liver - cytology; Liver - immunology; Liver - metabolism; Liver - physiopathology; Liver Development; Macrophages - chemistry; Macrophages - immunology; Mice; Mice, Inbred C57BL; Microscopy, Confocal; Microvessels - metabolism; Monocytes - chemistry; Mouse Model; Myeloid Cells - chemistry; Myeloid Cells - physiology; Phenotype; Transcriptome; APAP; clodronate-loaded liposomes; CLL; IL; indocyanine green; BM; ICG; green fluorescent protein; bone marrow; GFP; Kupffer cell; acetaminophen; time-of-flight mass cytometry; Liver Development; KC; HSC; hepatic stellate cell; interleukin; natural killer cell; CyTOF; NK; dendritic cell; DC; Mouse Model
• ISSN: 0016-5085; 1528-0012
• DOI: 10.1053/j.gastro.2016.08.024

Background & Aims 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. Methods We collected and analyzed liver tissues from C57BL/6 (control), LysM-EGFP, B6 ACTb-EGFP, CCR2 −/− , CD11c-EYFP, CD11c-EYFP-DTR, germ-free mice, CX3CR1 gfp/gfp , CX3CR1 gpf/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. Results 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. Conclusions 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.