Unique Proteomic Signatures Distinguish Macrophages and Dendritic Cells

Monocytes differentiate into heterogeneous populations of tissue macrophages and dendritic cells (DCs) that regulate inflammation and immunity. Identifying specific populations of myeloid cells in vivo is problematic, however, because only a limited number of proteins have been used to assign cellular phenotype. Using mass spectrometry and bone marrow-derived cells, we provided a global view of the proteomes of M-CSF-derived macrophages, classically and alternatively activated macrophages, and GM-CSF-derived DCs. Remarkably, the expression levels of half the plasma membrane proteins differed significantly in the various populations of cells derived in vitro. Moreover, the membrane proteomes of macrophages and DCs were more distinct than those of classically and alternatively activated macrophages. Hierarchical cluster and dual statistical analyses demonstrated that each cell type exhibited a robust proteomic signature that was unique. To interrogate the phenotype of myeloid cells in vivo, we subjected elicited peritoneal macrophages harvested from wild-type and GM-CSF-deficient mice to mass spectrometric and functional analysis. Unexpectedly, we found that peritoneal macrophages exhibited many features of the DCs generated in vitro. These findings demonstrate that global analysis of the membrane proteome can help define immune cell phenotypes in vivo

The enteroinsular axis in glucose dependent insulinotropic polypeptide receptor knockout (GIPR-/-) miee

The incretins, glucose-dependent insulinotropic polypeptide (GIP) and glucagonlike peptide-1 (GLP-1), are gut hormones that act via the enteroinsular axis to potentiate insulin secretion from the pancreas in a glucose-dependent manner. Both GLP-1 and GIP receptor knockout mice (GLP-1 R-/- and GIPR-/- respectively) have been generated to investigate the physiological importance of this axis. Studies in this thesis were carried out on GIP receptor knockout mice (GIPR-/-). Although reduced GIP action is a component of type 2 diabetes, GIP receptor-deficient mice exhibit only moderately impaired glucose tolerance. Thus, the present thesis was directed at investigating possible compensatory mechanisms that take place within the enteroinsular axis in the absence of GIP action. Fasting and 20t h minute OGTT serum GIP levels as well as duodenojejunal GIP content were altered in GIPR-/- mice. Total serum GLP-1 levels and serum DPIV activity in GIPR knockout mice were not significantly different from those in control animals, either before or during a glucose tolerance test. However, insulin responses to GLP-1 in pancreas perfusions and static islet incubations were significantly greater in GIPR -/- than in +/+ mice (P<0.05), and GLP-1 induced cAMP production was also elevated in the pancreatic islets of the knockout animals (P<0.05). Additionally, pancreatic insulin content and insulin gene expression were reduced in GIPR-/- mice compared to wild type (+/+) mice (P< 0.05). There was, however, no discernible difference in GLP-1 receptor mRNA levels. Immunohistochemistry studies revealed a normal distribution and localization of endocrine cells within the pancreatic islets of GIPR-/- mice. Surprisingly, these studies showed an increase in islet area, when compared to total pancreatic area, in the -/- mice (P<0.05) with less intense staining for insulin. In conclusion, the GIPR-/- mouse exhibits increased islet size and (3 cell sensitivity to GLP-1 despite a decrease in pancreatic insulin protein content and gene expression. These findings suggest a critical role for GIP in normal islet and (3 cell function and development.Arts, Faculty ofPsychology, Department ofGraduat

Geochemistry and mineralogy approaches to characterize brick and its lake sediments sources: Antioch Roman City (Southern Turkey)

The Roman aqueduct of Antioch-on-the-Orontes (Southern Turkey) is situated close to the Antioch city. This last is located near the Amik Lake (Lake of Antioch) and close to the junction between the active Dead Sea fault and the East Anatolian fault. During the Roman period, the Amik Plain was more densely occupied than at any time in its history [1]. The study focuses on the bricks and the lake sediments characterization in order to determine the source area as well as the technical production used at this period. For this purpose, several bricks were sampled on different parts of the city's aqueducts. Furthermore, a core of about 6 m of sediments was also collected from the dried Amik Lake. The bricks were characterized through a mineralogical (XRD) and chemical (PIXE-PIGE) approaches. Unfired clay fraction remained as inclusion in the brick was separated and then analysed using XRD. Geochemical composition and clay mineralogy were performed on the raw sediments from the Amik Lake in order to compare the source area. Technological test will be performed on the raw clay sediments from the Amik Lake in the purpose to understand the production techniques used at this time. The age of the brick production was previously dated to the Roman Period [2]. The synthesis of all the data attested the Amik Lake sediment as the raw material for the bricks of the aqueduct. Clay mineral composition from the Roman period deposited in the lake is smectite, illite, kaolinite and small amount of mixed-layer clays. The similar clays composition is found in the remained clays on the brick used for the aqueduct construction. Fast and heterogeneous firing practice characterized the manufacturing of these materials due to the rapid need for the materials during the post-seismic repairs after earthquakes that are mentioned in historical written works. [1] J. Casana, Geomorphology, 101, 429-442 (2008) [2] Y. Benjelloun, J. de Sigoyer, J. Carlut, A. Hubert-Ferrari, H. Dessales, H. Pamir, V. Karabacak, Comptes Rendus Geoscience, 347, 170-180 (2015

High triglyceride to HDL cholesterol ratio is associated with increased coronary heart disease among White but not Black adults

Objective: Black adults are less likely than White adults to present with adverse lipid profiles and more likely to present with low-grade inflammation. The impact of race on the association between atherogenic lipid profiles, inflammation, and coronary heart disease (CHD) is unknown. Methods: We evaluated the association between high levels (>50th percentile) of high-sensitivity C-reactive protein (hsCRP) and of triglycerides to high density lipoprotein ratio (TG/HDL-C) and CHD events by race in the REasons for Geographic and Racial Differences in Stroke (REGARDS) cohort with 30,239 Black and White participants aged 45 and older. Results: Participants with both high hsCRP and high TG/HDL-C had highest rates of CHD (HR 1.84; 95% CI: 1.48, 2.29 vs HR 1.52; 95% CI: 1.19, 1.94 in White vs Black participants respectively). Whereas isolated high hsCRP was associated with increased CHD risk in both races (HR 1.68; 95% CI: 1.31, 2.15 and HR 1.43; 95% CI: 1.13, 1.81 for White and Black participants respectively), isolated high TG/HDL was associated with increased CHD risk only in White participants (HR 1.44; 95% CI: 1.15, 1.79 vs HR 1.01; 95% CI: 0.74, 1.38). Further, the effects of high hsCRP and high TG/HDL-C were additive, with inflammation being the driving variable for the association in both races. Conclusion: In both races, higher inflammation combined with adverse lipid profile is associated with greater CHD risk. Therefore, inflammation increases CHD risk in both races whereas dyslipidemia alone is associated with a greater risk in White but not in Black adults. hsCRP testing should be a standard feature of CHD risk assessment, particularly in Black patients

Plasma membrane protein signatures of myeloid cells identify unique cell functions.

<p>Gene ontology analysis of plasma membrane proteins enriched in M1 macrophages, M2 macrophages, all macrophage types (BmM, M1, and M2), and BmDCs identifies functional categories of proteins enriched in each cell type (<i>p</i><0.05 with Benjamini-Hochberg correction). The top three functional annotations are presented for each cell type along with three representative proteins.</p

Immunocytochemical detection of plasma membrane protein markers.

<p>Expression levels of widely used plasma membrane protein markers (<b><i>Panels A–B</i></b>) and newly identified markers (<b><i>Panels C–D</i></b>) of M1 cells, M2 cells, BmMs, and BmDCs were assessed by mass spectrometry (<b><i>Panels A,C</i></b>) and immunocytochemistry (<b><i>Panels B,D</i></b>). For MS/MS, proteins were quantified by spectral counting and expressed relative to the cell type with the highest expression level for each protein. Results are means and SDs. Cells were stained with antibodies specific to each protein (red channel), counterstained with DAPI to visualize nuclei (blue-channel), and examined by confocal microscopy. Immunostaining and microscopy were performed on the same day with identical microscope settings. Results are representative of 3 independent analyses.</p