PXD008406

PXD008406 is an original dataset announced via ProteomeXchange.

Title	Expression of Siglec-E alters the proteome of LPS-activated macrophages but does not affect LPS-driven cytokine production or TLR4 endocytosis Expression of Siglec-E alters the proteome of LPS-activated macrophages but does not affect LPS-driven cytokine production or TLR4 endocytosis
Description	Siglec-E is a murine CD33-related siglec that functions as an inhibitory receptor and is expressed mainly on neutrophils and macrophage populations. Recent studies have suggested that siglec-E is an important negative regulator of LPS-TLR4 signalling and one report (Wu et al 2016) claimed that siglec-E is required for TLR4 endocytosis following uptake of Escherichia coli by macrophages and dendritic cells (DCs). Our attempts to reproduce these observations using cells from wildtype (WT) and siglec E deficient mice were unsuccessful. We used a variety of assays to determine if siglec-E expressed by different macrophage populations can regulate TLR-4 signalling in response to LPS, but found no consistent differences in cytokine secretion in vitro and in vivo, comparing 3 different strains of siglec-E-deficient mice with matched WT controls. No evidence was found that the siglec-E deficiency was compensated by expression of siglecs-F and –G, the other murine inhibitory CD33-related siglecs. Quantitative proteomics was used as an unbiased approach and provided additional evidence that siglec-E does not suppress inflammatory TLR4 signaling. Interestingly, proteomics revealed a siglec E dependent alteration in macrophage phenotype that could be relevant to functional responses in host defence. In support of this, siglec-E-deficient mice exhibited enhanced growth of Salmonella enterica serovar Typhimurium in the liver following intravenous infection, but macrophages lacking siglec-E did not show altered uptake or killing of bacteria in vitro. Using various cell types including bone marrow derived DCs (BMDC), splenic DCs and macrophages from WT and siglec-E-deficient mice, we showed that siglec-E is not required for TLR4 endocytosis following E.coli uptake or LPS challenge. We failed to see expression of siglec-E by BMDC even after LPS-induced maturation, but confirmed previous studies that splenic DCs express low levels of siglec-E. Taken together, our findings do not support a major role of siglec-E in regulation of TLR4 signalling functions or TLR4 endocytosis in macrophages or DCs. Instead, they reveal that induction of siglec-E by LPS can modulate the phenotype of macrophages, the functional significance of which is currently unclear.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:36:30.220.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Manjula Nagala
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-12-11 03:13:47	ID requested
1	2018-02-09 11:40:48	announced
⏵ 2	2024-10-22 04:36:38	announced	2024-10-22: Updated project metadata.

10.3389/fimmu.2017.01926;

Nagala M, McKenzie E, Richards H, Sharma R, Thomson S, Mastroeni P, Crocker PR, Expression of Siglec-E Alters the Proteome of Lipopolysaccharide (LPS)-Activated Macrophages but Does Not Affect LPS-Driven Cytokine Production or Toll-Like Receptor 4 Endocytosis. Front Immunol, 8():1926(2017) [pubmed]

curator keyword: Biological

submitter keyword: mouse BMDM LC-MSMS

Paul R crocker, Manjula Nagala
contact affiliation	Paul R crocker: Professor of Glycoimmunology and Head of the Division of Cell Signalling and Immunology
contact email	p.r.crocker@dundee.ac.uk
lab head
Manjula Nagala
contact affiliation	Research Associate
contact email	mnagala@dundee.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD008406
     1. Label: PRIDE project
     2. Name: Expression of Siglec-E alters the proteome of LPS-activated macrophages but does not affect LPS-driven cytokine production or TLR4 endocytosis Expression of Siglec-E alters the proteome of LPS-activated macrophages but does not affect LPS-driven cytokine production or TLR4 endocytosis