Biology subjects

Pajk, S.

Publications and source records attributed to Pajk, S..

3 recordsLinked to original sources

bioRxiv2025

Probing immune signatures of conjugated pattern recognition receptor ligands identifies chimeras with adjuvant and antitumor activity

Pattern recognition receptor (PRR) ligands hold great promise as adjuvants and immunotherapeutics. Here, we demonstrate that chemical conjugation of PRR ligands results in synergistic immune response amplification inaccessible to unlinked agonist mixtures. To identify potent immune agonists, we synthesized conjugated PRR ligands incorporating distinct agonist pairings, each targeting two carefully selected PRRs. We used a phenotypic screen using human peripheral blood mononuclear cells (PBMCs) to single out chimeric PRR ligands capable of inducing robust immune response both in terms of cytokine response and cytotoxicity against cancer cells. Chimeric TLR4/TLR7 and TLR7/RIG-I ligands showed broad immune activation in vitro as well as enhancement of antigen-specific cellular and humoral responses in mice. Intratumoral delivery of chimeric TLR4/TLR7 ligand induced robust antitumor response in a syngeneic mouse B16F10 tumor model. These results demonstrate the profound effects that conjugation can have on immune response and support the use of conjugated PRR ligands as adjuvants/immunotherapeutics. ONE SENTENCE SUMMARYConjugated PRR ligands enhance immune activation, serve as potent adjuvants, and induce antitumor response in a mouse model. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=97 SRC="FIGDIR/small/647694v1_ufig1.gif" ALT="Figure 1"> View larger version (15K): org.highwire.dtl.DTLVardef@d2904corg.highwire.dtl.DTLVardef@1ffa772org.highwire.dtl.DTLVardef@14a3592org.highwire.dtl.DTLVardef@103320a_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2021

Controlled Fluorescent Labelling of Metal Oxide Nanoparticles for Artefact-free Live Cell Microscopy

Nanotechnologies hold great promise for various applications. To predict and guarantee the safety of novel nanomaterials, it is essential to understand their mechanism of action in an organism, causally connecting adverse outcomes with early molecular events. They are best investigated using non-invasive advanced optical methods, such as high-resolution live-cell fluorescence microscopy, which require stable labelling of nanoparticles with fluorescent dyes. When performed inadequately, unbound fluorophores and inadvertently altered chemical and physical properties of the nanoparticles can, however, result in experimental artefacts and erroneous conclusions. To prevent such unintentional errors, we here describe a minimal combination of experimental methods to enable artefact-free fluorescent labelling of metal-oxide nanoparticles - the largest subpopulation of nanoparticles by industrial production and applications - and demonstrate its application in the case of TiO2 nanotubes. We 1) characterize potential changes of the nanoparticles surface charge and morphology that might occur during labelling, and 2) assess stable binding of the fluorescent dye to nanomaterial, which ensures correct nanoparticle localization. Together, these steps warrant the reliability and reproducibility of advanced optical tracking, which is necessary to explore nanomaterials mechanism of action and will foster widespread and safe use of new nanomaterials. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=182 SRC="FIGDIR/small/440400v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@358ffborg.highwire.dtl.DTLVardef@7faaecorg.highwire.dtl.DTLVardef@1ae7fb3org.highwire.dtl.DTLVardef@141c4b5_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

bioRxiv2020

Towards predictive nanotoxicology: from roundabout of molecular events to chronic inflammation prediction

Nanomaterial-induced diseases cannot be reliably predicted because of the lack of clearly identified causal relationships, in particular between acute exposures and chronic symptoms. By applying advanced microscopies and omics to in vitro and in vivo systems, together with in silico molecular modelling, we have here determined that the long-lasting response to a single exposure originates in the counteracting of a newly discovered nanomaterial quarantining and nanomaterial cycling among different lung cell types. This allows us to predict the nanomaterial-induced spectrum of lung inflammation using only in vitro measurements and in silico modelling. Besides its profound implications for cost-efficient animal-free predictive toxicology, our work also paves the way to a better mechanistic understanding of nanomaterial- induced cancer, fibrosis, and other chronic diseases.

molecular biology↗