A5026262726- T-cell and B-cell Immunology
- Cell Adhesion Molecules Research
- Advanced Fluorescence Microscopy Techniques
- Monoclonal and Polyclonal Antibodies Research
- Immune Cell Function and Interaction
- Cellular transport and secretion
- Complement system in diseases
- Caveolin-1 and cellular processes
- Cell Image Analysis Techniques
- Immunotherapy and Immune Responses
- CAR-T cell therapy research
- Cellular Mechanics and Interactions
- Glycosylation and Glycoproteins Research
- Lipid Membrane Structure and Behavior
- Phagocytosis and Immune Regulation
- Cytomegalovirus and herpesvirus research
- Galectins and Cancer Biology
- Calcium signaling and nucleotide metabolism
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Single-cell and spatial transcriptomics
- Atherosclerosis and Cardiovascular Diseases
- Hippo pathway signaling and YAP/TAZ
- Spectroscopy Techniques in Biomedical and Chemical Research
- Erythrocyte Function and Pathophysiology
- HIV Research and Treatment
Biotechnology Institute Thurgau
2018-2023
EMBL Australia
2016-2022
University of Konstanz
2018-2022
UNSW Sydney
2010-2022
ARC Centre of Excellence in Advanced Molecular Imaging
2015-2022
Australian Research Council
2016
Centenary Institute
2014
The University of Sydney
2014
University of Bern
2005-2011
The authors employed photoactivatable localization microscopy (PALM) and direct stochastic optical reconstruction (dSTORM) imaging image analysis based on Ripley's K-function to quantify the distribution heterogeneity of proteins at cell plasma membrane. membrane targeting sequence N-terminal region T receptor-pathway kinase Lck fused photo-convertible fluorescent protein tdEos (Lck(N10)-tdEos), clusters into sub-100 nm regions which cover approximately 7% surface. 2-channel PALM...
Antigen recognition by the T-cell receptor (TCR) is a hallmark of adaptive immune system. When TCR engages peptide bound to restricting major histocompatibility complex molecule (pMHC), it transmits signal via associated CD3 complex. How extracellular antigen event leads intracellular phosphorylation remains unclear. Here, we used single-molecule localization microscopy quantify organization TCR-CD3 complexes into nanoscale clusters and distinguish between triggered nontriggered complexes....
Abstract Tumor-specific T cells are frequently exhausted by chronic antigenic stimulation. We here report on a human antigen-specific ex vivo model to explore new therapeutic options for cell immunotherapies. generated with this resemble tumor-infiltrating phenotypic and transcriptional level. Using targeted pooled CRISPR-Cas9 screen individual gene knockout validation experiments, we uncover sorting nexin-9 (SNX9) as mediator of exhaustion. Upon TCR/CD28 stimulation, deletion SNX9 in CD8...
Abstract Endocytosis of surface receptors and their polarized recycling back to the plasma membrane are central many cellular processes, such as cell migration, cytokinesis, basolateral polarity epithelial cells T activation. Little is known about mechanisms that control organization endosomes how they connect receptor endocytosis. Here, we follow endocytic journey (TCR), from internalization at immunological synapse. We show TCR triggering leads its rapid uptake through a...
Neutrophils polarize and migrate in response to chemokines. Different types of membrane microdomains (rafts) have been postulated be present rear front polarized leukocytes disruption rafts by cholesterol sequestration prevents leukocyte polarization. Reggie/flotillin-1 -2 are two highly homologous proteins that ubiquitously enriched detergent resistant membranes thought shape forming homo- hetero-oligomers. It was the goal this study investigate dynamic microdomain reorganization during...
T cell signalling begins with the ligation of antigen receptor (TCR) by a cognate peptide and phosphorylation receptor's ITAM domains kinase Lck. However, canonical model is insufficient to explain how constitutively active Lck can discriminate between non-ligated ligated TCRs. Here, we discuss factors that are thought regulate spatial distribution TCR Lck, therefore critically influence initiation.
Significance Cell-expressed integrins mediate adhesion with other cells and extracellular matrix are essential for embryonic development controlling leukocyte migration in later life. Integrin depends on conformational change leading to activation, although it remains unknown exactly how alter their state response guidance cues. We show that the molecule plexinD1 controls clustering of patches cell membrane activation individual these can be switched off by binding sema3E plexinD1....
Objective— Therapeutic interventions that increase plasma levels of high-density lipoproteins and apolipoprotein A-I (apoA-I) A-I, the major lipoprotein apolipoprotein, improve glycemic control in people with type 2 diabetes mellitus. High-density apoA-I also enhance insulin synthesis secretion isolated pancreatic islets clonal β-cell lines. This study identifies signaling pathways mediate these effects. Approach Results— Incubation increased cAMP accumulation Ins-1E cells a...
Clustering of the T-cell receptor (TCR) is thought to initiate downstream signalling. However, detection protein clustering with high spatial and temporal resolution remains challenging. Here we establish a Förster resonance energy transfer (FRET) sensor, named CliF, which reports intermolecular associations neighbouring proteins in live cells. A key advantage single-chain FRET sensor that it can be combined image correlation spectroscopy (ICS), single-particle tracking (SPT) fluorescence...
Abstract Background Different types of membrane microdomains (rafts) have been postulated to be present in the rear and front polarized migrating T-lymphocytes. Disruption rafts by cholesterol sequestration prevents T-cell polarization migration. Reggie/flotillin-1 -2 are two highly homologous proteins that thought shape microdomains. We previously demonstrated enrichment flotillins uropod human neutrophils. now investigated mechanisms involved chemokine-induced flotillin reorganization...
Abstract Background The visualization of viral proteins has been hindered by the resolution limit conventional fluorescent microscopes, as dimension any single signal is often greater than most virion particles. Super-resolution microscopy potential to unveil distribution at approaching electron without relying on morphological features existing characteristics biological specimen that are needed in EM. Results Using direct stochastic optical reconstruction (dSTORM) achieve a lateral 15–20...
Annexin A6 (AnxA6) has been implicated in cell signalling by contributing to the organisation of plasma membrane. Here we examined whether AnxA6 regulates and proliferation T cells. We used a contact hypersensitivity model immune challenge wild‐type (WT) −/− mice found that vivo CD4 + cells, but not CD8 was impaired relative WT mice. However, T‐cell migration through receptor ex similar between cells isolated from In contrast, interleukin‐2 (IL‐2) reduced compared with Further,...
Abstract The targeted endocytic recycling of the T cell receptor (TCR) to immunological synapse is essential for activation. Despite this, mechanisms that underlie sorting internalised receptors into endosomes remain poorly understood. To build a comprehensive picture TCR during activation, we developed suite new imaging and quantification tools centred on photoactivation fluorescent proteins. We show membrane-organising proteins, flotillin-1 -2, are required reach Rab5-positive immediately...
Abstract Background T cell activation leads to increased expression of the receptor for iron transporter transferrin (TfR) provide required differentiation and clonal expansion that takes place during days after encounter with a cognate antigen. However, cells mobilise TfR their surface within minutes activation, although reason mechanism driving this process remain unclear. Results Here we show transiently increase endocytic uptake recycling upon thereby boosting capacity import iron. We...