A5083913528- Force Microscopy Techniques and Applications
- T-cell and B-cell Immunology
- Advanced Fluorescence Microscopy Techniques
- CAR-T cell therapy research
- Monoclonal and Polyclonal Antibodies Research
- Immunotherapy and Immune Responses
- Advanced biosensing and bioanalysis techniques
- Mechanical and Optical Resonators
- Immune Cell Function and Interaction
- Nanowire Synthesis and Applications
- Advancements in Semiconductor Devices and Circuit Design
- Cellular Mechanics and Interactions
- Cell Adhesion Molecules Research
- RNA Research and Splicing
- Nanofabrication and Lithography Techniques
- Advanced Electron Microscopy Techniques and Applications
- Cell Image Analysis Techniques
- Alcoholism and Thiamine Deficiency
- Platelet Disorders and Treatments
- Galectins and Cancer Biology
- Inhalation and Respiratory Drug Delivery
- Medical Imaging and Pathology Studies
- Silk-based biomaterials and applications
- Interstitial Lung Diseases and Idiopathic Pulmonary Fibrosis
- Telomeres, Telomerase, and Senescence
Medical University of Vienna
2011-2024
TU Wien
2016-2022
Max Perutz Labs
2011-2014
Johannes Kepler University of Linz
2013
Colorado State University
2011
Alternative splicing (AS) is an important regulatory process that leads to the creation of multiple RNA transcripts from a single gene. often carry premature termination codons (PTCs), which trigger nonsense-mediated decay (NMD), cytoplasmic degradation pathway. However, intron retention, most prevalent AS event in plants, PTC-carrying splice variants are insensitive NMD; this led us question fate these special variants. Here, we present innovative approach monitor and characterize...
Mechanical forces acting on ligand-engaged T-cell receptors (TCRs) have previously been implicated in antigen recognition, yet their magnitude, spread, and temporal behavior are still poorly defined. We here report a FRET-based sensor equipped either with TCR-reactive single chain antibody fragment or peptide-loaded MHC, the physiological TCR-ligand. The was tethered to planar glass-supported lipid bilayers (SLBs) informed most directly magnitude kinetics of TCR-imposed at molecule level....
Telomeres comprise the protective caps of natural chromosome ends and function in suppression DNA damage signaling cellular senescence. Therefore, techniques used to determine telomere length are important a number studies, ranging from those investigating telomeric structure effects on human disease. Terminal restriction fragment (TRF) analysis has for long time shown be one most accurate methods quantification absolute range species. As this technique centers standard Southern blotting, is...
Receptor–ligand interactions at cell interfaces initiate signaling cascades essential for cellular communication and effector functions. Specifically, T receptor (TCR) with pathogen-derived peptides presented by the major histocompatibility complex (pMHC) molecules on antigen-presenting cells are crucial activation. The binding duration, or dwell time, of TCR–pMHC correlates downstream efficacy, strong agonists exhibiting longer lifetimes compared to weak agonists. Traditional surface...
We present the software platform 2CALM that allows for a comparative analysis of 3D localisation microscopy data representing protein distributions in two biological samples. The in-depth statistical reveals differences between samples at nanoscopic level using parameters such as cluster-density and -curvature. An automatic classification system combines multiplex multi-level approaches into one comprehensive parameter similarity testing compared demonstrated importance 2CALM, comparing CD41...
Single molecule Förster resonance energy transfer (smFRET) is a popular tool to study biological systems that undergo topological transitions on the nanometer scale. smFRET experiments typically require recording of long trajectories and subsequent statistical analysis extract parameters such as states’ lifetimes. Alternatively, probability distributions exploits shapes at well chosen exposure times hence works without acquisition time traces. Here, we describe variant utilizes tests compare...
ABSTRACT Mechanical forces acting on ligand-engaged T-cell receptors (TCRs) have previously been implicated in antigen recognition, yet their magnitude, spread, and temporal behavior are still poorly defined. We here report a FRET-based sensor equipped with TCR-reactive single chain antibody fragment, which was tethered to planar supported lipid bilayers (SLBs) informs most directly the magnitude kinetics of TCR-imposed at molecule level. When confronting T-cells gel-phase SLBs we observed...
Receptor–ligand interactions at cell interfaces initiate signaling cascades essential for cellular communication and effector functions. Specifically, T receptor (TCR) with pathogen-derived peptides presented by the major histocompatibility complex (pMHC) molecules on antigen-presenting cells are crucial activation. The binding duration, or dwell time, of TCR–pMHC correlates downstream efficacy, strong agonists exhibiting longer lifetimes compared to weak agonists. Traditional surface...
Abstract Mechanical forces acting on ligand-engaged T-cell receptors (TCRs) have previously been implicated in antigen recognition and ligand discrimination, yet their magnitude, frequency, impact remain unclear. We quantitatively assess across various TCR:pMHC pairs with different bond lifetimes at single-molecule resolution, both before during activation, platforms that either include or exclude tangential force registration. Our results imply CD4+ TCRs experience significantly lower than...
We present a cluster spatial analysis method using nanoscopic dSTORM images to determine changes in protein distributions within brain tissue. Such methods are suitable investigate human tissue and will help achieve deeper understanding of disease along with aiding drug development. Human samples usually treated postmortem via standard fixation protocols, which established clinical laboratories. Therefore, our localization microscopy-based was adapted characterize density fixed different...
Interstitial lung disease comprises numerous clinical entities posing significant challenges towards a prompt and accurate diagnosis. Amongst the contributing factors are intricate pathophysiological mechanisms, an overlap between conditions, interobserver disagreement. We developed model for patient clustering offering additional approach to such complex cases. The is based on surface phenotyping of over 40 markers immune cells isolated from bronchoalveolar lavage in combination with data....
ABSTRACT Molecular crowding of agonist peptide/MHC class II complexes (pMHCIIs) with structurally similar, yet per se non-stimulatory endogenous pMHCIIs has been postulated to sensitize T-cells for the recognition single antigens on surface dendritic cells and B-cells. When testing this premise use advanced live cell microscopy, we observed as monomeric, randomly distributed entities diffusing rapidly after entering APC surface. Synaptic TCR-engagement highly abundant was low or non-existent...
A multitude of different imaging systems are already available to image genetically altered RNA species; however, only a few these techniques actually suitable visualize endogenous RNA. One possibility is use fluorescently-labelled and hybridization-sensitive probes. In order yield more information about the exact localization movement single molecule, it necessary such probes with highly sensitive microscope setups. More challenges arise if experiments conducted in plant cells due their...
ABSTRACT T-cell antigen receptors (TCRs) exhibit inherent cross-reactivity which broadens the spectrum of epitopes that are recognizable by a finite TCR-repertoire but also carries risk autoimmunity. However, TCRs support high level specificity as they allow T-cells to discriminate single antigenic peptide/MHC complexes (pMHCs) against millions structurally related self-pMHCs, in some cases based on absence or presence methyl-group. How manage convey such seemingly contrary properties and...
Low antigen sensitivity and a gradual loss of effector functions limit the clinical applicability chimeric receptor (CAR)–modified T cells call for alternative designs effective cell–based cancer immunotherapy. Here, we applied advanced microscopy to demonstrate that TCR/CD3-based synthetic constructs (TCC) outperform second-generation CAR formats with regard conveyed sensitivities by up thousandfold. TCC-based recognition occurred without adverse nonspecific signaling, which is typically...
Single-molecule Förster resonance energy transfer (smFRET) is a versatile technique reporting on distances in the sub-nanometer to nanometer range. It has been used wide range of biophysical and molecular biological experiments, including measurement forces, characterization conformational dynamics biomolecules, observation intracellular colocalization proteins, determination receptor-ligand interaction times. In widefield microscopy configuration, experiments are typically performed using...
Single-molecule Förster resonance energy transfer (smFRET) is a versatile technique reporting on distances in the sub-nanometer to nanometer range. It has been used wide range of biophysical and molecular biological experiments, including measurement forces, characterization conformational dynamics biomolecules, observation intracellular colocalization proteins, determination receptor-ligand interaction times. In widefield microscopy configuration, experiments are typically performed using...
ABSTRACT Low antigen sensitivity and a gradual loss of effector functions limit the clinical applicability chimeric receptor (CAR)-modified T-cells call for alternative designs effective T-cell-based cancer immunotherapy. Here we applied advanced microscopy to demonstrate that TCR/CD3-based synthetic constructs (TCC) outperform second-generation CAR formats with regard conveyed sensitivities by up thousand-fold. TCC-based recognition occurred without adverse non-specific signaling, which is...