Klaus Eyer

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Eyer

First Name

Klaus

ORCID

0000-0001-9344-5110

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Publications 1 - 10 of 58

Deep phenotypic characterization of immunization-induced antibacterial IgG repertoires in mice using a single-antibody bioassay
Heo, Millie; Chenon, Guilhem; Castrillon, Carlos; et al. (2020)
Communications Biology
Antibodies with antibacterial activity need to bind to the bacterial surface with affinity, specificity, and sufficient density to induce efficient elimination. To characterize the anti-bacterial antibody repertoire, we developed an in-droplet bioassay with single-antibody resolution. The assay not only allowed us to identify whether the secreted antibodies recognized a bacterial surface antigen, but also to estimate the apparent dissociation constant (KD app) of the interaction and the density of the recognized epitope on the bacteria. Herein, we found substantial differences within the KD app/epitope density profiles in mice immunized with various species of heat-killed bacteria. The experiments further revealed a high cross-reactivity of the secreted IgG repertoires, binding to even unrelated bacteria with high affinity. This application confirmed the ability to quantify the anti-bacterial antibody repertoire and the utility of the developed bioassay to study the interplay between bacteria and the humoral response. © 2020, The Author(s).
A microfluidic vesicle screening platform
Kuhn Phillip; Eyer, Klaus; Allner, Steffen; et al. (2011)
Analytical Chemistry
Quantitative modeling of the effect of antigen dosage on B-cell affinity distributions in maturating germinal centers
Molari, Marco; Eyer, Klaus; Baudry, Jean; et al. (2020)
eLife
Affinity maturation is a complex dynamical process allowing the immune system to generate antibodies capable of recognizing antigens. We introduce a model for the evolution of the distribution of affinities across the antibody population in germinal centers. The model is amenable to detailed mathematical analysis and gives insight on the mechanisms through which antigen availability controls the rate of maturation and the expansion of the antibody population. It is also capable, upon maximum-likelihood inference of the parameters, to reproduce accurately the distributions of affinities of IgG-secreting cells we measure in mice immunized against Tetanus Toxoid under largely varying conditions (antigen dosage, delay between injections). Both model and experiments show that the average population affinity depends non-monotonically on the antigen dosage. We show that combining quantitative modeling and statistical inference is a concrete way to investigate biological processes underlying affinity maturation (such as selection permissiveness), hardly accessible through measurements.
Dynamic Activation of NADPH Oxidases in Immune Responses Modulates Differentiation, Function, and Lifespan of Plasma Cells
Bucheli, Olivia T.M.; Rodrigues, Daniela; Ulbricht, Carolin; et al. (2025)
European Journal of Immunology
NADPH-oxidase (NOX)-derived reactive oxygen species (ROS) have been described to play essential roles in B-cell activation processes. However, several key questions concerning NOX activity and subsequent ROS production remain unaddressed, including fundamental processes such as differentiation, functional competence, cellular metabolism, and viability. This study investigated these questions in a murine B-cell response after secondary immunization. We combined single-cell transcriptomics and single-cell detection of NOX activity and observed that various subsets of B cells dynamically express NOX1 and NOX2. The NOX+ cellular phenotype correlated with increased activity of metabolic pathways, augmented lactate production, lower IgG secretion rates, and markers for longevity. The NOX+ cellular phenotype was also associated with increased cellular stress and apoptosis, underscoring the intricate relationship between ROS and cellular survival. Consequently, these insights advance our understanding of how long-lived humoral immunity is formed.
Rapid estimation of the energy charge from cell lysates using matrix-assisted laser desorption/ionization mass spectrometry
Steinhoff, Robert F.; Krismer, Jasmin; Eyer, Klaus; et al. (2014)
Analytical Biochemistry
Antibody density on bacteria regulates C1q recruitment by monoclonal IgG but not IgM
Aymerich, Nathan; Schlotheuber, Luca J.; Bucheli, Olivia T.M.; et al. (2024)
European Journal of Immunology
Antibodies that trigger the complement system play a pivotal role in the immune defense against pathogenic bacteria and offer potential therapeutic avenues for combating antibiotic-resistant bacterial infections, a rising global concern. To gain a deeper understanding of the key parameters regulating complement activation by monoclonal antibodies, we developed a novel bioassay for quantifying classical complement activation at the monoclonal antibody level, and employed this assay to characterize rare complement-activating antibacterial antibodies on the single-antibody level in postimmunization murine antibody repertoires. We characterized monoclonal antibodies from various antibody isotypes against specific pathogenic bacteria (Bordetella pertussis and Neisseria meningitidis) to broaden the scope of our findings. We demonstrated activation of the classical pathway by individual IgM- and IgG-secreting cells, that is, monoclonal IgM and IgG2a/2b/3 subclasses. Additionally, we could observe different epitope density requirements for efficient C1q binding depending on antibody isotype, which is in agreement with previously proposed molecular mechanisms. In short, we found that antibody density most crucially regulated C1q recruitment by monoclonal IgG isotypes, but not IgM isotypes. This study provides additional insights into important parameters for classical complement initiation by monoclonal antibodies, a knowledge that might inform antibody screening and vaccination efforts.
A new mechanobiological era
Kurth, Felix; Eyer, Klaus; Franco-Obregón, Alfredo; et al. (2012)
Current Opinion in Chemical Biology
A facile protocol for the immobilisation of vesicles, virus particles, bacteria, and yeast cells
Kuhn, Philipp; Eyer, Klaus; Robinson, Tom; et al. (2012)
Integrative Biology
Full Spectroscopic Tip-Enhanced Raman Imaging of Single Nanotapes Formed from β-Amyloid(1–40) Peptide Fragments
Paulite, Melissa; Blum, Carolin; Schmid, Thomas; et al. (2013)
ACS Nano
Detection of Infertility-related Neutralizing Antibodies with a Cell-free Microfluidic Method
Eyer, Klaus; Root, Katharina; Verboket, Pascal E.; et al. (2015)
Scientific Reports
The unwanted emergence of neutralizing antibodies (nAbs) against an endogenous or a therapeutic protein can result in deficiency diseases or therapy failure. Here, we developed a cell-free microfluidic method for the sensitive detection and quantification of nAbs in human serum that are associated with infertility. We used cell-derived vesicles containing the luteinizing hormone (LH)/choriogonadotropin receptor (LHHCGR) to detect nAbs against LH. The method exploits the entire cellular signal amplification mechanism, and facilitates the detection of as little as 0.44 nM of LH-nAb (Kd 1.5 nM) in human serum matrix within only 15 minutes. In addition, dose-response curves can be generated in less than 2 hours to evaluate the nAB concentration and dissociation constant. The developed system is devoid of problems associated with cell-based assays and we believe that this simple effect-directed analysis can be used in clinical environments, and is adaptable to other hormones or cytokines and their respective nAbs.

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Klaus Eyer

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