Biology subjects

Van Kasteren, S. I.

Publications and source records attributed to Van Kasteren, S. I..

3 recordsLinked to original sources

bioRxiv2025

A Novel Cyclopropenyl Fatty Acid Library Reveals Tissue-Specific Preferences for Regulatory T Cell Uptake Through Click-Chemistry

The activation of T-cells is heavily shaped by the nutrients that are available during activation. Fatty acids can have highly pleiotropic effects in this process. On the one hand, they are essential for driving T-cell activation, yet on the other hand they can affect curtailed T-cell activation. These differences are likely dependent on the nature and absolute uptake of the fatty acids. Quantifying the uptake of specific FAs by individual cells and studying the effect on the phenotype of the cell is currently not possible with existing tools. Here we therefore use the live-cell compatible Inverse Electron-Demand Diels-Alder reaction combined with the synthesis of saturated, unsaturated and polyunsaturated fatty acids carrying the 1-carbon cyclopropene click group. Single cell uptake studies of these various clickable FAs in primary immune cell mixtures show highly divergent uptake behaviour between different immune cells, with polyunsaturated fatty acids markedly preferred by all immune cells tested.

immunology↗

bioRxiv2025

Glyco-PAINT-APP: Subcellular analysis of immune cell-lectin binding enables correlation of glycan binding parameters to receptor biology

Extracting single-molecule lectin binding kinetics from primary cells has not been possible to date. Here, we present Glyco-PAINT-APP (Automated Processing Pipeline), an automated method that enables the extraction of subcellular glycan interaction kinetics using a Points Accumulation for Imaging in Nano-Topography (PAINT)-based approach. This approach leverages an algorithm for precise, high-throughput subcellular analysis of glycan binding dynamics, facilitating the exploration of functional correlations between glycoform binding patterns and immune cell polarization. Using synthetic glycans and glycosylated antigens, we demonstrate the ability of the technique to automatically correlate glycan binding parameters in subregions of dendritic cell membranes with increased uptake and cross-presentation efficiency of these antigens. Additionally, we show how the method can uncover subtle differences in glycan binding preferences between resting and polarized macrophages. Taken together, Glyco-PAINT-APP has the potential to enable new insights into the cell-intrinsic heterogeneity of glycan-structure-activity relationships in immune cells.

cell biology↗

bioRxiv2022

QUAS-R: Glutamine (Q) Uptake Assay with Single cell Resolution reveals metabolic heterogeneity with immune populations.

System level analysis of single cell data is rapidly transforming the field of immunometabolism. However, metabolic profiling of single cells and small populations by flow and mass cytometry is extremely limited by the availability of specific reagents such as antibodies and fluorescently nutrient analogues. Given the competitive demand for nutrients in pathogenic microenvironments including sites of infection, tumours and autoinflammation, there is a need to understand how and when immune cells access these nutrients. Fluorescent-tagging of nutrients is one approach to study nutrient transport but is extremely limited in its usefulness as tagging usually changes the transport characteristics and transporter specificity of the nutrient. Herein, we developed a completely new approach for single cell analysis of nutrient uptake where a fluorophore is attached to a functionalized amino acid after it has been transported across the plasma membrane and is within the cell. This in-cell biorthogonal labelling ensures that bona fide transport has been measured. System ASC transporter SLC1A5/ASCT2 transports multiple amino acids, most notably the crucial fuel glutamine, and has essential roles in supporting immune metabolism, signalling and function. This flow cytometry assay allows for rapid, sensitive, and quantitative measurement of SLC1A5-mediated uptake, which we used to interrogate the transport capacity of the complex immune subpopulations within the thymus, at a single cell resolution previously "unreachable". Taken together, our findings provide an easy procedure to assess which cells support their function via SLC1A5 mediated uptake of amino acids in a sensitive single cell assay. This assay is a significant addition to the single-cell metabolic toolbox required to decode the metabolic landscape of complex immune microenvironments.

immunology↗