Biology subjects

Geier, F.

Publications and source records attributed to Geier, F..

4 recordsLinked to original sources

bioRxiv2025

Non-cytolytic re-engineering of a viral vaccine vector enables durable effector-memory T cell immunity by reinforcing type I IFN induction

Replication-deficient viral vector systems hold promise for CD8 T cell-based vaccination, but the molecular mechanisms accounting for platform-specific differences in immunogenicity remain ill- defined. When comparing prototypic single-cycle vaccine delivery platforms we found that lymphocytic choriomeningitis virus-based vectors (rLCMV), which are non-cytolytic, elicited more durable and effector-memory-differentiated CD8 T cell responses than vectors based on cytolytic vesicular stomatitis virus (rVSV). Hence we re-engineered rVSV to be non-cytolytic (rVSVMq). This vector induced more durable and effector-differentiated CD8 T cell memory than the parental rVSV and it afforded superior protection against Listeria challenge. Improved CD8 T cell responses of non-cytolytic rVSVMq were driven by a reinforced type I interferon (IFN-I) response and its direct sensing by vaccination-induced CD8 T cells. Many vector cargo-specific CD8 T cells in the splenic marginal zone of rVSVMq- or rLCMV-vaccinated mice were in contact with vector cargo-expressing cells that co- expressed type I interferon. In contrast, rVSV-vectored cargo-expressing contacts of specific CD8 T cells were largely IFN type I-negative. Thereby, vaccination with non-cytolytic viral vectors offered an opportunity for CD8 T cells to integrate peptide-MHC and IFN-I signals during priming. These mechanistic insights should help to refine vaccines aimed at eliciting durable and protective effector- memory CD8 T cell immunity.

immunology↗

bioRxiv2025

Identification and proteomic profiling of CD90+ small EVs using a refined immunocapture separation approach targeting stromal-derived EV subpopulations in synovial fluid of arthritis patients

Extracellular vesicles (EVs) have emerged as essential drivers in disease progression and promising biomarkers across various disease conditions, including autoimmune diseases such as rheumatoid arthritis (RA). RA is characterized by chronic inflammation driven by patient-specific cellular sub-phenotypes within the synovium. Since EVs reflect their cellular origin and are detectable in biofluids like synovial fluid, they offer a promising alternative for novel biomarker discovery or diagnostics in arthritis, potentially reducing the need for invasive tissue biopsies. However, the inherent complexity, viscosity and extracellular matrix (ECM) composition of synovial fluid pose significant challenges for efficient EV isolation. Moreover, most previous studies have focused on heterogenous bulk EV populations, neglecting the diverse EV subsets present in patient biofluids. To address these challenges, we established a novel immunocapture-based separation method, that selectively targets and isolates cell type-specific EV subpopulation from the heterogenous EV pool present in synovial fluid of arthritis patients. As a proof-of-concept approach, we targeted a stromal-derived EV population by leveraging the fibroblast-associated surface marker CD90/THY1, representing a subset of synovial fibroblasts implicated as important drivers in synovitis and chronic inflammation in RA. Western Blot and data independent acquisition (DIA) liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) proteomic analysis of immunocaptured CD90+ EVs isolated from cultured primary synovial fibroblasts and synovial fluid of arthritis patients, confirmed the successful separation of stromal vs a myeloid/lymphoid EV populations. The in-depth proteomic profiling of synovial fluid EVs offers unprecedented insights into EV heterogeneity in arthritis. This analysis enables the classification of cell-type associated EV subsets and the identification of novel EV markers for future immunocapture-based EV separation strategies. Our refined fit-for-purpose EV separation approach, coupled to in depth proteomic profiling, provides a powerful tool to identify disease-relevant EV subpopulations and novel biomarker candidates in synovial fluid, facilitating advances in arthritis diagnostics and personalized medicine

immunology↗

bioRxiv2021

Defective Desmosomal Adhesion Causes Arrhythmogenic Cardiomyopathy by involving an Integrin-αVβ6/TGF-β Signaling Cascade

BackgroundArrhythmogenic Cardiomyopathy (ACM) is characterized by progressive loss of cardiomyocytes with fibrofatty replacement, systolic dysfunction and life-threatening arrhythmias. A substantial proportion of ACM is caused by mutations in genes of the desmosomal cell-cell adhesion complex, but the underlying mechanisms are not well understood. So far, treatment options are only symptomatic. Here, we investigate the relevance of defective desmosomal adhesion for ACM development and progression. MethodsWe mutated the binding site of desmoglein-2 (DSG2), a crucial desmosomal adhesion molecule in cardiomyocytes. This DSG2-W2A mutation abrogates the tryptophan swap, a central interaction mechanism of DSG2 based on structural data. Impaired adhesive function of DSG2-W2A was confirmed by cell-cell dissociation assays and force spectroscopy measurements by atomic force microscopy. We next generated a DSG2-W2A knock-in mouse model, which was analyzed by echocardiography and histological and bio-molecular techniques including RNA sequencing, transmission electron and super-resolution microscopy. The results were compared to ACM patient samples and their relevance was confirmed in cardiac slice cultures. ResultsThe DSG2-W2A mutation induced impaired binding and desmosomal adhesion dysfunction on cellular and molecular level. Mice bearing this mutation develop a severe cardiac phenotype recalling the characteristics of ACM, including cardiac fibrosis, impaired systolic function and arrhythmia. A comparison of the transcriptome of mutant mice with ACM patient data suggested deregulated integrin-V{beta}6 and subsequent TGF-{beta} signaling as driver of cardiac fibrosis. Accordingly, blocking antibodies targeting integrin-V{beta}6 or inhibition of TGF-{beta} receptor signaling both led to reduced expression of pro-fibrotic markers in cardiac slice cultures. ConclusionsHere, we show that disruption of desmosomal adhesion is sufficient to induce ACM, which confirms the dysfunctional adhesion hypothesis. Mechanistically, deregulation of integrin-V{beta}6 signaling was identified as a central step towards fibrosis. This highlights the value of this model to discern mechanisms of cardiac fibrosis and to identify and test novel treatment options for ACM.

cell biology↗

bioRxiv2019

Persistence and plasticity of T follicular memory cells

CD4 memory T cells play an important role in protective immunity and are a key target in vaccine development. Many studies have focused on T central memory (TCM) cells, while the existence and functional significance of T follicular helper (TFH) memory cells is controversial. Here we show that TFH memory cells are highly susceptible to NAD induced cell death (NICD) during isolation from tissues, leading to their under-representation in prior studies. NICD blockade reveals the persistence of abundant TFH memory cells, with high expression of hallmark TFH markers, that persist to at least 400 days after infection, by which time TCM cells are no longer found. Using single cell RNA-seq we demonstrate that TFH memory cells are transcriptionally distinct from TCM cells, maintain stemness and self-renewal gene expression, and, in contrast to TCM cells, are multipotent following recall. Surprisingly, TFH memory cells concurrently express a distinct glycolytic signature similar to trained immune cells, including elevated expression of mTOR, HIF-1 and cAMP regulated genes. Late disruption of glycolysis/ICOS signaling leads to TFH memory cell depletion concomitant with decreased splenic plasma cells and circulating antibody titers, demonstrating both unique homeostatic regulation of memory TFH and their sustained function during the memory phase of the immune response. These results highlight the metabolic heterogeneity underlying distinct memory T cell subsets and establish TFH memory cells as an attractive target for the induction of long-lived adaptive immunity.\n\nHIGHLIGHTSO_LICell death during isolation from the tissue prevents the full recovery of TFH memory cells\nC_LIO_LITFH memory cells are transcriptionally distinct from TCM cells and maintain a broader recall capacity\nC_LIO_LITFH memory cells are maintained in the absence of antigen but require ICOS signaling and glycolysis\nC_LIO_LITFH memory cells support late phase antibody production by splenic plasma cells\nC_LI

immunology↗