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

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• 作者：Bruna Araujo David¹ ; Rafael Machado Rezende² ; Maí ; sa Mota Antunes¹ ; Mô ; nica Morais Santos¹ ; Maria Alice Freitas Lopes¹ ; Ariane Barros Diniz¹ ; Rafaela Vaz Sousa Pereira¹ ; Sarah Cozzer Marchesi¹ ; Dé ; bora Moreira Alvarenga¹ ; Brenda Naemi Nakagaki¹ ; Alan Moreira Araú ; jo¹ ; Daniela Silva dos Reis³ ; Renata Monti Rocha³ ; Pedro Elias Marques³ ; Woo-Yong Lee⁴ ; Justin Deniset⁴ ; Pei Xiong Liew⁴ ; Stephen Rubino² ; Laura Cox² ; Vanessa Pinho⁵ ; Thiago Mattar Cunha⁶ ; Gabriel Rocha Fernandes⁷ ; André ; Gustavo Oliveira¹ ; ⁸ ; Mauro Martins Teixeira³ ; Paul Kubes⁴ ; ^{pkubes@ucalgary.ca} ; Gustavo Batista Menezes¹ ; ^{menezesgb@ufmg.br}
• 关键词：CyTOF ; DC ; Liver Development ; Mouse Model
• 刊名：Gastroenterology
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：151
• 期：6
• 页码：1176-1191
• 全文大小：8487 K
• 卷排序：151

文摘

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.MethodsWe 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.ResultsUsing 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.ConclusionsIn 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.