Aims/hypothesisThe inflammatory milieu characteristic of insulitis affects translation fidelity and generates defective ribosomal products (DRiPs) that participate in autoimmune beta cell... Show moreAims/hypothesisThe inflammatory milieu characteristic of insulitis affects translation fidelity and generates defective ribosomal products (DRiPs) that participate in autoimmune beta cell destruction in type 1 diabetes. Here, we studied the role of early innate cytokines (IFNα) and late immune adaptive events (IFNɣ) in insulin DRiP-derived peptide presentation to diabetogenic CD8+ T cells.MethodsSingle-cell transcriptomics of human pancreatic islets was used to study the composition of the (immuno)proteasome. Specific inhibition of the immunoproteasome catalytic subunits was achieved using siRNA, and antigenic peptide presentation at the cell surface of the human beta cell line EndoC-βH1 was monitored using peptide-specific CD8 T cells.ResultsWe found that IFNγ induces the expression of the PSMB10 transcript encoding the β2i catalytic subunit of the immunoproteasome in endocrine beta cells, revealing a critical role in insulin DRiP-derived peptide presentation to T cells. Moreover, we showed that PSMB10 is upregulated in a beta cell subset that is preferentially destroyed in the pancreases of individuals with type 1 diabetes.Conclusions/interpretationOur data highlight the role of the degradation machinery in beta cell immunogenicity and emphasise the need for evaluation of targeted immunoproteasome inhibitors to limit beta cell destruction in type 1 diabetes. Show less
Nowadays, therapeutic antibodies are the major and fastest growing class of biotherapeuticals. Since their invention, they are continuously developed to improve structural and functional... Show moreNowadays, therapeutic antibodies are the major and fastest growing class of biotherapeuticals. Since their invention, they are continuously developed to improve structural and functional characteristics. The high complexity of recently generated antibody derivatives, with various modifications induced during the manufacturing process itself leads to many proteoform variants of the desired product. These proteoforms can potentially exhibit altered activity. Therefore, an adequate characterization of the proteoforms, the assessment of their impact and careful monitoring of critical species is indispensable in order to guarantee effective and safe biopharmaceuticals. As the landscape of next-generation Ab formats continuously evolves, it is likewise of great importance to further develop appropriate analytical methods for their thorough attribute analysis. Hence, the focus of the research performed in this thesis is the development of multi-level approaches for the in-depth, primarily MS-based characterization of biopharmaceuticals to overcome the present restrictions and challenges arising e.g. by the implementation of complex Ab formats. Show less
As a non-invasive genetic method, eDNA based approaches have become an important component of ecologists' and environmental managers' toolkits for biomonitoring in conservation and an increasingly... Show moreAs a non-invasive genetic method, eDNA based approaches have become an important component of ecologists' and environmental managers' toolkits for biomonitoring in conservation and an increasingly important source of ecological knowledge. This thesis focuses on aquatic eDNA based approaches beyond detections of species presence by 1) enhancing the knowledge of the characteristics of aquatic eDNA particle size distribution (PSD), 2) exploring the possibility of eDNA analysis in physiological investigation and 3) performing functional gene analysis across species. Aquatic eDNA PSD changes with degradation interacting with environmental factors and species. This knowledge supports the capability of eDNA PSD analysis in assessing false- and real-positive of species presence, as well as contributing to abundance assessment and guiding sampling strategy development when using eDNA based approaches. eDNA methylation varies between life stages, and supports life stage evaluations, and brings up the possibility of using eDNA methylation analysis in assessing physiological information. eDNA based approaches support functional gene analysis across species. Functional gene analysis promotes investigating ARGs pollution and the interaction of pollution between different types of agricultural regions. Thanks to the continuously increasing insights on environment-related gene function and expression level, it becomes potentially possible to use eDNA functional gene (and methylation) analysis to evaluate environmental changes. Overall, this thesis contributes to expanding the application of aquatic eDNA based approaches beyond detections of species presence, and provides efficient and non-invasive approaches for biomonitoring and ecological assessments. Show less
