A5012332000- Light effects on plants
- Immune Cell Function and Interaction
- CAR-T cell therapy research
- Transgenic Plants and Applications
- T-cell and B-cell Immunology
- Photoreceptor and optogenetics research
- Photosynthetic Processes and Mechanisms
- Immunodeficiency and Autoimmune Disorders
- Receptor Mechanisms and Signaling
- Galectins and Cancer Biology
- Circadian rhythm and melatonin
- RNA Interference and Gene Delivery
- Viral Infectious Diseases and Gene Expression in Insects
- Nanowire Synthesis and Applications
- Monoclonal and Polyclonal Antibodies Research
- Chronic Lymphocytic Leukemia Research
- CRISPR and Genetic Engineering
- Photochromic and Fluorescence Chemistry
- Mast cells and histamine
- Olfactory and Sensory Function Studies
- bioluminescence and chemiluminescence research
- Single-cell and spatial transcriptomics
- Biotin and Related Studies
- SARS-CoV-2 and COVID-19 Research
- Virus-based gene therapy research
University of Freiburg
2014-2024
University Medical Center Freiburg
2016-2024
Massachusetts Institute of Technology
2019
Max Planck Society
2014
RWTH Aachen University
2013
The immune system distinguishes between self and foreign antigens. kinetic proofreading (KPR) model proposes that T cells discriminate from ligands by the different ligand binding half-lives to cell receptor (TCR). It is challenging test KPR as available experimental systems fall short of only altering keeping other parameters interaction unchanged. We engineered an optogenetic using plant photoreceptor phytochrome B (PhyB) a selectively control dynamics TCR light. This opto-ligand-TCR was...
The adapter protein linker for activation of T cells (LAT) is a critical signaling hub connecting cell antigen receptor triggering to downstream responses. In this study, we describe the first kindred with defective LAT caused by homozygous mutation in exon 5, leading premature stop codon deleting most cytoplasmic tail LAT, including tyrosine residues signal propagation. three patients presented from early childhood combined immunodeficiency and severe autoimmune disease. Unlike mouse...
Activation of the T cell receptor (TCR) by antigen is key step in adaptive immunity.In abTCR, induces a conformational change at CD3 subunits (CD3 CC) that absolutely required for abTCR activation.Here, we demonstrate CC not induced stimulation mouse G8 or human Vg9Vd2 gdTCR.We find there fundamental difference between activation mechanisms and gdTCR map to constant regions TCRab/gd heterodimers.Enforced induction with less commonly used monoclonal anti-CD3 promoted proximal signaling but...
Optogenetics is a versatile and powerful tool for the control analysis of cellular signaling processes. The activation receptors by light using optogenetic switches usually requires genetic manipulation cells. However, this considerably limits application in primary, nonengineered cells, which crucial study physiological processes controlling cell fate function therapeutic purposes. To overcome limitation, we developed system light-dependent extracellular surface cells termed OptoREACT...
Optogenetic approaches have gathered momentum in precisely modulating and interrogating cellular signalling gene expression. The use of optogenetics on the outer cell surface to interrogate how cells receive stimuli from their environment, however, has so far not reached its full potential. Here we demonstrate development an optogenetically regulated membrane receptor-ligand pair exemplified by optically responsive interaction integrin receptor with extracellular matrix. system is based...
Methodologies for the controlled delivery of genetic information into target cells are utmost importance engineering in both fundamental and applied research. However, available methods efficient gene transfer user-selected or even single suffer from low throughput, need complicated equipment, high invasiveness, side effects by off-target viral uptake. Here, we engineer an adeno-associated (AAV) vector system that transfers native upon illumination with cell-compatible red light. This...
Ligand binding to the TCR causes a conformational change at CD3 subunits expose CD3ε cytoplasmic proline-rich sequence (PRS). It was suggested that PRS is important for signaling and T cell activation. has been shown purified, recombinant SH3.1 domain of adaptor molecule noncatalytic region tyrosine kinase (Nck) can bind exposed CD3ε, but molecular mechanism how full-length Nck binds in cells not investigated so far. Using situ proximity ligation assay copurifications, we show requires...
Activation of mast cells (MCs) can be achieved by the high-affinity receptor for IgE (FcεRI) as well additional receptors such lipopolysaccharide (LPS) and tyrosine kinase Kit (stem cell factor [SCF] receptor). Thus, pharmacological interventions which stabilize MCs in response to different would preferable diseases with pathological systemic MC activation mastocytosis. 1,4-Benzodiazepines (BDZs) have been reported suppress effector functions. In present study, our aim was analyze...
Multiprotein complexes control the behavior of cells, such as lymphocytes immune system. Methods to affinity purify protein and determine their interactome by mass spectrometry are thus widely used. One drawback these methods is presence false positives. In fact, elution interest (POI) done changing biochemical properties buffer, so that unspecifically bound proteins (the positives) may also elute. Here, we developed an optogenetics-derived light-controlled purification method based on...
In the field of extracellular optogenetics, photoreceptors are applied outside cells to obtain systems with a desired functionality. Among diverse photoreceptors, phytochromes only ones that can be actively and reversibly switched between active inactive photostate by illumination cell-compatible red far-red light. this protocol, we describe production biotinylated variant photosensory domain A. thaliana phytochrome B (PhyB-AviTag) in E. coli single, optimized expression plasmid. We give...
T cells are one major cell type of the immune system that use their antigen receptor (TCR) to bind and respond foreign molecules derived from pathogens. The ligand-TCR interaction half-lives determine stimulation outcome. Until recently, scientists relied on mutating either TCR or its ligands investigate how varying TCR-ligand durations impacted activation. Our newly created opto-ligand-TCR allowed us precisely reversibly control ligand binding by light illumination. This uses phytochrome B...
The kinetics of a ligand-receptor interaction determine the responses receptor-expressing cell. One approach to experimentally and reversibly change this on demand is optogenetics. We have previously developed system in which modified receptor with an engineered ligand can be controlled by light. In soluble Phytochrome B (PhyB) tetramer fused mutated PhyB-interacting factor (PIF S ). However, often natural not soluble, but expressed as membrane protein another This allows interactions two...
Activation of T cells by agonistic peptide-MHC can be inhibited antagonistic ones. However, the exact mechanism remains elusive. We used Jurkat expressing two different TCRs and tested whether stimulation endogenous TCR anti-Vβ8 antibodies modulated ligand-binding to second, optogenetic TCR. The latter uses phytochrome B tetramers (PhyBt) as ligand, binding half-life which altered light. show that this determined PhyBt acted a second agonist (long half-life), an antagonist (short half-life)...
Optogenetics offers a set of tools for the precise manipulation signaling pathways. Here we exploit optogenetics to experimentally change kinetics protein-protein interactions on demand. We had developed system in which interaction modified T cell receptor (TCR) with an engineered ligand can be controlled by light. The was plant photoreceptor phytochrome B (PhyB) and TCR included TCRβ chain fused GFP mutated PhyB-interacting factor (PIFS), resulting GFP-PIFS-TCR. failed engineer...
Abstract The pivotal task of the immune system is to distinguish between self and foreign antigens. kinetic proofreading model (KPR) proposes that T cells discriminate from ligands by different ligand binding half-lives cell receptor (TCR). It challenging test KPR as available experimental systems fall short only altering keeping other parameters ligand-TCR interaction unchanged. We engineered an optogenetic using plant photoreceptor phytochrome B selectively control dynamics TCR light....
Abstract SARS-CoV-2, the causative agent of Covid-19, is known to evade immune system by several mechanisms. This includes shutdown host cellular protein synthesis, which abrogates induction antiviral interferon responses. The virus initiates infection susceptible cells binding with its spike (S) angiotensin-converting enzyme 2 (ACE2). Here we applied T cell receptor fusion construct (TRuC) technology engineer against such infected cells. In our TRuCs an S-binding domain fused CD3ε component...