scholarly journals Spatiotemporal proteomic profiling of the pro-inflammatory response to lipopolysaccharide in the THP-1 human leukaemia cell line

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Claire M. Mulvey ◽  
Lisa M. Breckels ◽  
Oliver M. Crook ◽  
David J. Sanders ◽  
Andre L. R. Ribeiro ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Proteome Analysis ◽  
Subcellular Location ◽  
Interferon Response ◽  
Subcellular Localisation ◽  
Proteomic Profiling ◽  
Leukaemia Cell Line ◽  
Physiological Processes ◽  
Intracellular Compartments ◽  
Almost All

AbstractProtein localisation and translocation between intracellular compartments underlie almost all physiological processes. The hyperLOPIT proteomics platform combines mass spectrometry with state-of-the-art machine learning to map the subcellular location of thousands of proteins simultaneously. We combine global proteome analysis with hyperLOPIT in a fully Bayesian framework to elucidate spatiotemporal proteomic changes during a lipopolysaccharide (LPS)-induced inflammatory response. We report a highly dynamic proteome in terms of both protein abundance and subcellular localisation, with alterations in the interferon response, endo-lysosomal system, plasma membrane reorganisation and cell migration. Proteins not previously associated with an LPS response were found to relocalise upon stimulation, the functional consequences of which are still unclear. By quantifying proteome-wide uncertainty through Bayesian modelling, a necessary role for protein relocalisation and the importance of taking a holistic overview of the LPS-driven immune response has been revealed. The data are showcased as an interactive application freely available for the scientific community.

scholarly journals Quantitative Proteome Analysis of Atg5-Deficient Mouse Embryonic Fibroblasts Reveals the Range of the Autophagy-Modulated Basal Cellular Proteome

mSystems ◽  
2019 ◽  
Vol 4 (6) ◽  
Author(s):  
Kiran Bala Sharma ◽  
Manish Sharma ◽  
Suruchi Aggarwal ◽  
Amit Kumar Yadav ◽  
Shinjini Bhatnagar ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Inflammatory Response ◽  
Transforming Growth Factor ◽  
Proteome Analysis ◽  
Interferon Response ◽  
Type I ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
The Impact

ABSTRACT Basal autophagy is crucial for maintenance of cellular homeostasis. ATG5 is an essential protein for autophagosome formation, and its depletion has been extensively used as a tool to disrupt autophagy. Here, we characterize the impact of Atg5 deficiency on the cellular proteome of mouse embryonic fibroblasts (MEFs). Using a tandem mass tagging (TMT)-based quantitative proteomics analysis, we observe that 14% of identified proteins show dysregulated levels in atg5−/− MEFs. These proteins were distributed across diverse biological processes, such as cell adhesion, development, differentiation, transport, metabolism, and immune responses. Several of the upregulated proteins were receptors involved in transforming growth factor β (TGF-β) signaling, JAK-STAT signaling, junction adhesion, and interferon/cytokine-receptor interactions and were validated as autophagy substrates. Nearly equal numbers of proteins, including several lysosomal proteins and enzymes, were downregulated, suggesting a complex role of autophagy/ATG5 in regulating their levels. The atg5−/− MEFs had lower levels of key immune sensors and effectors, including Toll-like receptor 2 (TLR2), interferon regulatory factor 3 (IRF3), IRF7, MLKL, and STAT1/3/5/6, which were restored by reexpression of ATG5. While these cells could efficiently mount a type I interferon response to the double-stranded RNA (dsRNA) mimic poly(I·C), they were compromised in their inflammatory response to the bacterial pathogen-associated molecular patterns (PAMPs) lipopolysaccharide (LPS) and Pam3CSK4. Transcriptional activation and secretion of interleukin-6 (IL-6) in these cells could be recovered by ATG5 expression, supporting the role of autophagy in the TLR2-induced inflammatory response. This study provides a key resource for understanding the effect of autophagy/ATG5 deficiency on the fibroblast proteome. IMPORTANCE Autophagy performs housekeeping functions for cells and maintains a functional mode by degrading damaged proteins and organelles and providing energy under starvation conditions. The process is tightly regulated by the evolutionarily conserved Atg genes, of which Atg5 is one such crucial mediator. Here, we have done a comprehensive quantitative proteome analysis of mouse embryonic fibroblasts that lack a functional autophagy pathway (Atg5 knockout). We observe that 14% of the identified cellular proteome is remodeled, and several proteins distributed across diverse cellular processes with functions in signaling, cell adhesion, development, and immunity show either higher or lower levels under autophagy-deficient conditions. These cells have lower levels of crucial immune proteins that are required to mount a protective inflammatory response. This study will serve as a valuable resource to determine the role of autophagy in modulating specific protein levels in cells.

scholarly journals Ocular Autonomic Nervous System: An Update from Anatomy to Physiological Functions

Vision ◽  
2022 ◽  
Vol 6 (1) ◽  
pp. 6
Author(s):  
Feipeng Wu ◽  
Yin Zhao ◽  
Hong Zhang
Keyword(s):  
Nervous System ◽  
Autonomic Nervous System ◽  
Neural Control ◽  
Entire Body ◽  
Physiological Functions ◽  
Autonomic Nervous ◽  
Parasympathetic Nerves ◽  
Physiological Processes ◽  
New Research ◽  
Almost All

The autonomic nervous system (ANS) confers neural control of the entire body, mainly through the sympathetic and parasympathetic nerves. Several studies have observed that the physiological functions of the eye (pupil size, lens accommodation, ocular circulation, and intraocular pressure regulation) are precisely regulated by the ANS. Almost all parts of the eye have autonomic innervation for the regulation of local homeostasis through synergy and antagonism. With the advent of new research methods, novel anatomical characteristics and numerous physiological processes have been elucidated. Herein, we summarize the anatomical and physiological functions of the ANS in the eye within the context of its intrinsic connections. This review provides novel insights into ocular studies.

scholarly journals Modern views on the problem of sepsis

2020 ◽  
pp. 18-26
Author(s):  
V.S. Kopcha
Keyword(s):  
Infectious Disease ◽  
Inflammatory Response ◽  
Clinical Case ◽  
Conservative Therapy ◽  
Medical Community ◽  
Etiological Factors ◽  
Systemic Reactions ◽  
Almost All ◽  
Increased Activity

BACKGROUND. By defining the cornerstone of sepsis as the “systemic” of the inflammatory response in the form of cytokine violations, the authors of the corresponding concept are forced to recognize the etiological factors almost all pathogens of infectious diseases, which naturally go with the increased activity of cytokines in serum. Without diminishing the importance of these components of inflammation, one should pay attention to the postulate that not only viruses and protozoa never cause sepsis, but also all non-bacterial pathogens are not etiological factors of this disease, although they lead to severe systemic reactions accompanied by an increase in the level of proinflammatory cytokines. OBJECTIVE. The purpose of the work is to draw the attention of physicians to the discrepancy between the new notions about sepsis imposed on the medical community and accepted not only by anesthetists, resuscitative surgeons and surgeons, but also by many infectionists. MATERIALS AND METHODS. A detailed analysis of the clinical case of sepsis in a young woman is presented, which, due to adequate surgical and long-term conservative therapy, has safely recovered. CONCLUSIONS. Sepsis is a generalized acyclic infectious disease of the bacterial and/or fungal etiology that develops in an immunodeficient organism with characteristic pathomorphological and pathologist changes in organs and tissues.

scholarly journals Subcellular location and properties of bactericidal factors from human neutrophils.

1986 ◽  
Vol 164 (5) ◽  
pp. 1407-1421 ◽  
Author(s):  
J E Gabay ◽  
J M Heiple ◽  
Z A Cohn ◽  
C F Nathan
Keyword(s):  
Protein S ◽  
Subcellular Location ◽  
Inhibitory Effect ◽  
Human Neutrophils ◽  
E Coli ◽  
Fractionation Scheme ◽  
Percoll Gradients ◽  
Nonionic Detergent ◽  
Almost All ◽  
Triton X

We examined the subcellular location of bactericidal factors (BF) in human neutrophils, using an efficient fractionation scheme. Nitrogen bomb cavitates of DIFP-treated PMN were centrifuged through discontinuous Percoll gradients, each fraction extracted with 0.05 M glycine, pH 2.0, and tested for the killing of Escherichia coli. greater than 90% of BF coisolated with the azurophil granules. After lysis of azurophils, 98% of azurophil-derived BF (ADBF) sedimented with the membrane. ADBF activity was solubilized from azurophil membrane with either acid or nonionic detergent (Triton X-100, Triton X-114). Bactericidal activity was linear with respect to protein concentration over the range 0.3-30 micrograms/ml. 0.1-0.3 microgram/ml ADBF killed 10(5) E. coli within 30 min at 37 degrees C. At 1.4 micrograms/ml, 50% of 2 X 10(5) bacteria were killed within 5 min. ADBF was effective between pH 5-8, with peak activity at pH 5.5. Glucose (20 mM), EDTA (1-25 mM), and physiologic concentrations of NaCl or KCl had little or no inhibitory effect on ADBF. ADBF killed both Gram-positive and Gram-negative virulent clinical isolates, including listeria, staphylococci, beta-hemolytic streptococci, and Pseudomonas aeruginosa. Thus, under these conditions of cell disruption, fractionation, extraction, and assay, almost all BF in human PMN appeared to be localized to the membrane of azurophilic granules as a highly potent, broad-spectrum, rapidly acting protein(s) effective in physiologic medium. Some of these properties appear to distinguish ADBF from previously described PMN bactericidal proteins.

scholarly journals Sperm Proteome Analysis and Identification of Fertility-Associated Biomarkers in Unexplained Male Infertility

Genes ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 522 ◽  
Author(s):  
Selvam ◽  
Agarwal ◽  
Pushparaj ◽  
Baskaran ◽  
Bendou
Keyword(s):  
Male Infertility ◽  
Radical Scavenging ◽  
Surface Protein ◽  
Proteome Analysis ◽  
Fold Change ◽  
Semen Parameters ◽  
Proteomic Profiling ◽  
Altered Expression ◽  
Fertilization Process ◽  
Infertile Men

Up to 30% of men with normal semen parameters suffer from infertility and the reason for this is unknown. Altered expression of sperm proteins may be a major cause of infertility in these men. Proteomic profiling was performed on pooled semen samples from eight normozoospermic fertile men and nine normozoospermic infertile men using LC-MS/MS. Furthermore, key differentially expressed proteins (DEPs) related to the fertilization process were selected for validation using Western blotting. A total of 1139 and 1095 proteins were identified in normozoospermic fertile and infertile men, respectively. Of these, 162 proteins were identified as DEPs. The canonical pathway related to free radical scavenging was enriched with upregulated DEPs in normozoospermic infertile men. The proteins associated with reproductive system development and function, and the ubiquitination pathway were underexpressed in normozoospermic infertile men. Western blot analysis revealed the overexpression of annexin A2 (ANXA2) (2.03 fold change; P = 0.0243), and underexpression of sperm surface protein Sp17 (SPA17) (0.37 fold change; P = 0.0205) and serine protease inhibitor (SERPINA5) (0.32 fold change; P = 0.0073) in men with unexplained male infertility (UMI). The global proteomic profile of normozoospermic infertile men is different from that of normozoospermic fertile men. Our data suggests that SPA17, ANXA2, and SERPINA5 may potentially serve as non-invasive protein biomarkers associated with the fertilization process of the spermatozoa in UMI.

scholarly journals Proteome Analysis Reveals Distinct Mitochondrial Functions Linked to Interferon Response Patterns in Activated CD4+ and CD8+ T Cells

2019 ◽  
Vol 10 ◽  
Author(s):  
Marlene C. Gerner ◽  
Laura Niederstaetter ◽  
Liesa Ziegler ◽  
Andrea Bileck ◽  
Astrid Slany ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Proteome Analysis ◽  
Interferon Response ◽  
Response Patterns ◽  
Mitochondrial Functions

scholarly journals Gravin-associated kinase signaling networks coordinate γ-tubulin organization at mitotic spindle poles

2020 ◽  
Vol 295 (40) ◽  
pp. 13784-13797
Author(s):  
Paula J. Bucko ◽  
Irvin Garcia ◽  
Ridhima Manocha ◽  
Akansha Bhat ◽  
Linda Wordeman ◽  
...  
Keyword(s):  
Quantitative Imaging ◽  
Subcellular Location ◽  
Asymmetric Distribution ◽  
Binding Partner ◽  
Mitotic Kinases ◽  
Spindle Poles ◽  
Biochemical Assays ◽  
Anchoring Protein ◽  
Intracellular Compartments ◽  
Genetic Deletion

Mitogenic signals that regulate cell division often proceed through multienzyme assemblies within defined intracellular compartments. The anchoring protein Gravin restricts the action of mitotic kinases and cell-cycle effectors to defined mitotic structures. In this report we discover that genetic deletion of Gravin disrupts proper accumulation and asymmetric distribution of γ-tubulin during mitosis. We utilize a new precision pharmacology tool, Local Kinase Inhibition, to inhibit the Gravin binding partner polo-like kinase 1 at spindle poles. Using a combination of gene-editing approaches, quantitative imaging, and biochemical assays, we provide evidence that disruption of local polo-like kinase 1 signaling underlies the γ-tubulin distribution defects observed with Gravin loss. Our study uncovers a new role for Gravin in coordinating γ-tubulin recruitment during mitosis and illuminates the mechanism by which signaling enzymes regulate this process at a distinct subcellular location.

Proteomic profiling uncovered the cytosolic superoxide dismutase BsSOD1 associated with plant defence in the herbal orchid Bletilla striata

2020 ◽  
Vol 47 (10) ◽  
pp. 937
Author(s):  
Bao-Wei Lu ◽  
Feng-Xia An ◽  
Liang-Jing Cao ◽  
Yong-Jian Yang ◽  
Peng-Ming Liu ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Fungal Infection ◽  
Fluorescent Protein ◽  
Yellow Fluorescent Protein ◽  
Plant Defence ◽  
Subcellular Localisation ◽  
Proteomic Profiling ◽  
Oxidoreductase Activity ◽  
Bletilla Striata ◽  
Significant Gene

The herbal orchid Bletilla striata (Thunb.) Rchb.f. has a long cultivation history and has been widely used in medicines and cosmetics. The fungal infection leaf blight (LB) seriously threatens B. striata cultivation. Here, we systemically collected wild B. striata accessions and isolated the accessions with strong resistance against LB. We carried out proteomic profiling analysis of LB-resistant and LB-susceptible accessions, and identified a large number of differentially expressed proteins with significant gene ontology enrichment for ‘oxidoreductase activity.’ Of the proteins identified in the reactive oxygen species signalling pathway, the protein abundance of the Cu–Zn superoxide dismutase BsSOD1 and its gene expression level were higher in LB-resistant accessions than in LB-susceptible lines. Transient expression of the dismutase fused with yellow fluorescent protein determined that its subcellular localisation is in the cytoplasm. Our study provides new insights into the molecular markers associated with fungal infection in B. striata.

A Cellular Proteome Map of Human Multiple Myeloma.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 111-111 ◽  
Author(s):  
Johannes Drach ◽  
Astrid Slany ◽  
Verena Sagaster ◽  
Nina Gundacker ◽  
Verena Haudek ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Protein Synthesis ◽  
Er Stress ◽  
Proteome Analysis ◽  
Interferon Response ◽  
Patient Stratification ◽  
Aberrant Expression ◽  
Aberrant Cell ◽  
Human Multiple Myeloma ◽  
The Impact

Abstract Background and Aims: Molecular profiling identifies proteins characteristically deregulated in malignant diseases. Characteristic biomarkers may be useful to support diagnosis and patient stratification, while the recognition of aberrant cell activities and cell survival strategies may lead to the development of specifically designed pharmacologic strategies. We therefore performed proteomic profiling of primary human multiple myeloma (MM) cells in order to define the impact of protein expression abnormalities in MM. Methods: Plasma cells were isolated from bone marrow of patients with MM or MGUS and forwarded to proteome analysis based on 2D gel electrophoresis in addition to shotgun analysis by nano-LC-MS/MS. Erythrocytes, platelets and plasma as well as quiescent and activated lymphocytes, monocytes, endothelial and dendritic cells from healthy donors were processed in an identical manner for comparative analysis. The resulting data were interpreted with the aid of a homemade SQL database. Results: Among about thousand proteins identified in MM cells, we found aberrant expression of proteins involved in fatty acid beta-oxidation (VLCAD), unfolded protein response/ER-stress (ARMET protein, cytosol aminopeptidase), oxidative stress (ste20/oxidant stress responsive kinase 1), interferon response (MX1), iron uptake (transferrin receptor/CD71), DNA modification (transforming protein ERG, methyltransferase-like protein 7A), apoptosis and survival (apoptosis-inducing factor 1, hsp75), protein synthesis (ribosome-binding protein, Unc-13 D), cell adhesion (CD9/tetraspanin-29, LYRIC/metastatic adhesion protein), signaling (sts-1) and cell-cell interaction (Cystatin F, basigin/collagenase stimulatory factor, stem cell growth factor, small inducible cytokine B7, PD-ECGF/Gliostatin). Conclusion: The presently applied proteome analysis strategy, based on the systematic investigation of purified primary human cells, allowed us to find characteristic alterations in MM cells. Several proteins directly relate to known aberrant cell activities such as elevated protein synthesis and secretion resulting in ER-stress, which makes the cells sensitive to proteasome inhibitor treatment. Work is in progress to use quantitative assessment of such proteins for patient stratification, identification of response predictors, and biomarkers for the distinction of MM and MGUS.

Exosomes: Applications in Respiratory Infectious Diseases and Prospects for Coronavirus Disease 2019 (COVID-19)

2020 ◽  
Vol 16 (4) ◽  
pp. 399-418
Author(s):  
Jia Feng ◽  
Ahmed Waqas ◽  
Zhihan Zhu ◽  
Lukui Chen
Keyword(s):  
Infectious Diseases ◽  
Immune Surveillance ◽  
Cell Communication ◽  
Stem Cell Differentiation ◽  
Target Cells ◽  
Physiological Processes ◽  
Global Pandemic ◽  
New Ideas ◽  
Respiratory Infectious Diseases ◽  
Almost All

Exosomes are small extracellular vesicles of 30–150 nm diameter secreted by almost all cells. In recent years, with continuous deeper understanding of exosomes physiological functions, different reports have proven that exosomes can facilitate cell-to-cell communication by binding to target cells and transferring their contents, together with RNAs, DNAs, proteins, and lipids between cells and tissues. With advantages that exosomes can be involved in various types of physiological processes, such as blood coagulation, cellular homeostasis, inflammation, immune surveillance, stem cell differentiation, neuroprotection, and tissue regeneration and angiogenesis. Exosomes have been demonstrated that they can be applied in identification and treatment of multiple disorders such as cancers, cerebral ischemia, and respiratory infectious diseases. Importantly, researchers utilize application of exosomes in the treatment of various respiratory infectious diseases that have made some breakthrough progress. However, with the global pandemic of Coronavirus Disease 2019 (COVID-19), we have focused on applications of exosomes in respiratory infectious diseases and their serious complications, including influenza, TB, ARDS and sepsis. In this review, we explain the use of exosomes in various respiratory infectious diseases and their serious complications, and hope to provide new ideas for the treatment of new coronavirus infections.

Sign in / Sign up

Export Citation Format

Share Document