Dataset Information

Freshly isolated rat liver sinusoidal endothelial cells and Kupffer cells, LC-MSMS

ABSTRACT: Liver sinusoidal endothelial cells (LSECs) and Kupffer cells (KCs) have important roles in liver homeostasis and host defense. Sharing the same microenvironment in the liver sinusoid, they form an effective scavenger cell system for removal of potentially harmful blood-borne substances. Unlike most other endothelia, LSECs are highly efficient in endocytosis of nanoparticles, including virus. Though controversial, LSECs have been reported to act as antigen presenting cells, thus contributing importantly to induction of immune tolerance in liver. There are also controversies about LSEC and KC specific markers, which may be due to overlapping cell functions, species differences, and/or problems with cell purification. We therefore used label-free proteomics to characterize and quantitatively compare proteome of freshly isolated, highly pure rat LSECs (SE-1/CD32b positive) and KCs (CD11b/c positive.We found that most immune genes expressed in KCs were also expressed in LSECs, albeit at a lower density, and they also have overlap in cell surface marker expression. Both cell types express high levels of scavenger receptors and immune lectins.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Kupffer Cell, Endothelial Cell Of Sinusoid, Primary Cell, Liver

SUBMITTER: Sabin Bhandari

LAB HEAD: Karen Kristine Sørensen

PROVIDER: PXD012080 | Pride | 2020-11-11

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 15

Similar Datasets

Project description:Liver sinusoidal endothelial cells (LSECs) are specialized fenestrated scavenger endothelial cells involved in the elimination of modified plasma proteins and tissue turnover waste macromolecules from blood. LSECs also participate in liver immune responses. A challenge when studying LSEC biology is the rapid loss of the in vivo cell phenotype in culture. In this study, we have examined biological processes and pathways affected during early-stage primary culture of rat LSECs and checked for cell responses to the pro-inflammatory cytokine interleukin (IL)-1 and the anti-inflammatory drug dexamethasone. Methods: LSECs from male Sprague Dawley rats were cultured on type I collagen in a 5% oxygen atmosphere in DMEM with serum-free supplements for 2 and 24 h. Quantitative proteomics using tandem mass tag technology were used to examine proteins in cells and supernatants. Validation was done with ELISA and multiplex immunobead assays, cell ultrastructure was examined by scanning electron microscopy, and scavenger function by quantitative endocytosis assays. Results: LSECs cultured for 24 h showed a characteristic pro-inflammatory phenotype both in the presence and absence of IL-1, displaying upregulation of cellular responses to cytokines and interferon-, cell-cell adhesion, and glycolysis, and downregulation of membrane/endocytosis receptors (MCAM, STAB1, STAB2, LYVE1, CLEC4G) and proteins involved in pyruvate metabolism, citric acid cycle, fatty acid elongation, amino acid metabolism, and oxidation-reduction processes. Dexamethasone improved LSEC viability in culture and repressed the culture-induced stimulation of glycolysis, inflammatory and immune regulatory pathways, and secretion of pro-inflammatory cytokines while increasing IL-10 secretion compared to time-matched control. The number of sieve plates in LSECs was reduced at 24 h both in the presence and absence of dexamethasone but the dexamethasone-treated cells showed a more quiescent phenotype. Conclusion: Rat LSECs become activated towards a pro-inflammatory phenotype during early culture. Dexamethasone represses LSEC activation and improves cell viability.

Project description:The liver is located at the interface between the intestinal portal vasculature and the general circulation, employing a complex set of innate and adaptive immune cells to protect from the continuous systemic threat of gut-translocating pathogens. In contrast to lymphoid tissues in which asymmetric, polarized spatial positioning of distinct cell types plays an essential role in optimal host defense, the relationship between liver phagocytes and lymphoid element distribution and their immune functions is currently unknown. This lack of organization stands in marked contrast to the extensively documented zonation of hepatocyte metabolic function controlled by Wnt signaling during development. Here we used quantitative multi-parameter confocal imaging to re-examine resident immune cells distribution in the liver, finding that both myeloid (Kupffer cell -KC) and lymphoid (NKT cell) components are substantially enriched in the peri-portal regions of the liver lobule, the entry point for portal venous blood rich in gut-derived pathogens and molecules. Peri-portal immune enrichment of KCs was lost in germ-free (GF) animals and coincided temporally with weaning and microbial colonization, suggesting that immune cell positioning was not a developmentally programmed process but rather dependent on extrinsic factors. Conditional ablation of MyD88-dependent bacterial product sensing by liver sinusoidal endothelial cells (LSECs) disrupted KC spatial organization animals, consistent with immune cell positioning within the liver architecture arising from sustained, active responses of LSECs to microbial stimuli. Bacterial challenge studies revealed that this asymmetric peri-portal-biased immune cell localization protects the liver and the host against bacterial dissemination. Together, these data reveal that LSECs act as microbiome sensors, actively promoting concentration of protective immune components in the sub-regions of the liver most susceptible to infection to provide optimal host defense against systemic pathogen spread.

			Action	DRS
	170524_RL_N2.raw	Raw
	170524_RL_N2b.raw	Raw
	170524_RL_N3.raw	Raw
	170524_RL_N3b.raw	Raw
	170524_RL_N4.raw	Raw

Dataset Information

Freshly isolated rat liver sinusoidal endothelial cells and Kupffer cells, LC-MSMS

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets