Biology subjects

Hofmann, U.

Publications and source records attributed to Hofmann, U..

12 recordsLinked to original sources

bioRxiv2026

Ex vivo Infrared Nerve Stimulation on the Rat Sciatic Nerve: Challenges and Pitfalls

Infrared nerve stimulation (INS) offers millisecond-scale, electrical artifact-free activation of peripheral nerves, yet rat ex vivo studies remain scarce. We present an INS setup utilizing a lens system for free-beam focus for rat sciatic nerves partially submerged in a nerve bath, making re-wetting of the tissue in existing ex vivo INS setups redundant. Excised sciatic nerve preparations were illuminated at 1470 nm, delivering pulse trains from 100 - 2000 s at 5 Hz with radiant exposures up to 25.7 J/cm2. Compound action potentials (CAPs) were recorded with tungsten hook electrodes. Nerve activity could be recorded up to 200 minutes after excision, though persistent nerve excitability remained comparable to existing ex vivo studies as a yet to be optimized parameter. Peak CAP amplitudes ranged from 3.9 - 36.9 {micro}V with latencies averaged at 3.4 ms. Activation thresholds spanned 1.75 - 13.05 J/cm2. Additionally, potential pitfalls for INS setups and ways to resolve them were investigated, with two artifact types highly relevant to ex vivo INS being identified: A photo-thermal expansion artifact and a photovoltaic artifact with thermo-capacitive coupling. The observations confirm that CAPs can be evoked via INS in an ex vivo preparation of the rat sciatic nerve. The presented platform supports 3R principles and offers a robust basis for introduction of pharmacological investigations with INS.

neuroscience↗

bioRxiv2026

Treatment with saturating dose of conventional anti-CD28 monoclonal antibody well tolerated in pig acute myocardial infarction model

CD4+ Foxp3+ regulatory T cells (Treg) efficiently foster wound healing after myocardial infarction (MI). Therapeutically shifting the balance between CD4+ Foxp3- conventional T cells (Tconv) and Treg towards Tregs enhanced survival in a mouse model of MI. Due to species-specific differences in cardiac wound healing and remodelling, it remains, however, unclear whether these findings can be translated into novel immunotherapies for human patients after MI. Therefore, we studied pigs whose cardiac wound healing after MI and the composition of the immune system are very close to humans. This includes the relevant complication of developing a cytokine release syndrome (CRS) after infusion of saturating amounts of a superagonistic anti-CD28 monoclonal antibody (mAb). To achieve the intended shift in the Treg/Tconv balance, we treated pigs three days after interventional MI induction with a non-superagonistic, i.e. conventional, anti-CD28 mAb, clone 3D11. Infusion of a saturating dose (1 mg/kg body weight) of mAb 3D11 was clinically well tolerated without signs of CRS induction or any other complications. Molecularly, mAb 3D11 infusion led to a downmodulation of CD28 expression on porcine T cells in vivo with the remaining CD28 molecules blocked from binding natural ligand proteins CD80 and CD86, as we show in this publication. Apart from modulating CD28 expression, treatment with mAb 3D11 did not induce any overt changes in the peripheral T cell compartment. However, after mAb 3D11 treatment, we observed Treg accumulation in the infarcted heart, particularly the border zone, on day 7 post-MI using immunofluorescence histology. Our findings thus suggest that even saturating doses of conventional anti-CD28 monoclonal antibodies could potentially be safely administered in patients to therapeutically shift the Treg/Tconv balance in the infarcted myocardium. This might be sufficient to enhance cardiac wound healing in patients short-term and prevent adverse remodelling long-term.

immunology↗

bioRxiv2025

Changes in peripheral blood leukocyte composition precede development of heart-reactive autoantibodies in patients hospitalised for acute heart failure

In a retrospective pilot study, we showed that the induction of heart-reactive autoantibodies (HRA) in the wake of acutely decompensated heart failure predicts worse outcomes. To gain deeper insights into the immunological mechanisms causing induction of HRA after heart failure decompensation we initiated the prospective Acute Heart Failure-Immunomonitoring Cohort Study (AHF-ImmunoCS). For this study, 380 patients were enrolled and will be followed up, including serial collection of biomaterials, for a period of 18 months after the index hospitalisation for AHF. Analysis of AHF-ImmunoCS samples obtained at baseline and at 6-month follow-up from 110 patients showed de novo induction of HRA - as detected by indirect immunofluorescence (IFT) - in 21% of patients (previously published percentage: 32%). The IFT results did not reflect induction of broad anti-heart autoimmunity as autoantibodies against other cardiac antigens like Troponin I3 or Myosin Light Chain 7 were not induced in parallel. To understand what drives HRA induction in these patients we longitudinally immunophenotyped peripheral blood leukocytes at baseline, 6-week and 6-month follow-up by high-resolution spectral flow cytometry. Among lymphocytes, induction of HRA in the wake of acute decompensation of heart failure was associated with a higher proportion of CD4+ T cells among lymphocytes, more CD45RA+ CCR7+ naive conventional, i.e. non-regulatory, and more CXCR3+ CCR4- Th1 cells among CD4+ T cells at baseline. Among myeloid cells, there were no differences at baseline between patients going on to develop HRA and those that did not. However, patients developing HRA had higher proportions of eosinophils (six-month follow-up) and lower proportions of Arginase+ HLA-DR- polymorphnuclear myeloid-derived suppressor cells among myeloid cells (six-week and six-month follow-up). Our data, thus, implicate that alterations in the composition of both the lymphoid and the myeloid compartments might drive HRA induction which impacts disease progression and prognosis in AHF.

immunology↗

bioRxiv2025

CD4+ T cells fuel the Duchenne cardiomyopathy

Duchenne muscular dystrophy (DMD) is a X-linked genetic disorder, in which cardiomyopathy represents a major cause of mortality. Although myocardial inflammation and fibrosis are hallmarks of the disease, the role of adaptive immunity in cardiac pathology remains poorly defined. Using the Mdx mouse model, we demonstrate that T and B lymphocytes accumulate in the heart and that activation occurs in heart-draining lymph nodes at an early disease stage. Mdx mice lacking adaptive immunity (Mdx-Scid) were protected from myocardial fibrosis and hypertrophic remodeling. Single-cell RNA sequencing revealed expansion of an inflammatory, matrisome-associated macrophage subset in Mdx but not Mdx-Scid hearts. Genetic deficiency or depletion of CD4+ T cells reduced left ventricular fibrosis and preserved systolic function. Moreover, adoptive transfer of T cells von Mdx mice induced myocardial fibrosis and dysfunction in healthy recipients. Our results identify autoreactive CD4+ T cells as key drivers of DMD-associated cardiomyopathy and suggest targeted modulation of adaptive immune responses as a potential therapeutic approach in DMD.

immunology↗

bioRxiv2025

Low-dose infusion of a superagonistic anti-CD28 monoclonal antibody ameliorates inflammation after acute myocardial infarction in pigs

CD4+ Foxp3+ regulatory T cells (Treg) efficiently foster wound healing after myocardial infarction (MI). Moreover, therapeutic shifting of the balance between CD4+ Foxp3- conventional (Tconv) and Treg towards Treg enhances survival in mice after MI. Owing to the species-specific differences in cardiac wound healing and remodelling after MI, it remains unclear whether these findings can be translated into novel immunotherapies for patients with MI. Therefore, we tested the feasibility of therapeutic Treg activation after MI in pigs whose cardiac wound healing and the composition of the immune system are very close to humans. To activate Treg, we treated pigs three days after MI induction with a superagonistic anti-CD28 monoclonal antibody (mAb, CD28-SA; clone 4D12). To avoid induction of a cytokine release syndrome (CRS) we infused the CD28-SA at a low dose of 10 {micro}g/kg body weight. Low-dose CD28-SA treatment led to a local increase in porcine Treg among heart-infiltrating CD4+ T cells in comparison with the control mAb therapy (clone MOPC-21). Immunofluorescence histology showed that CD28-SA application particularly strengthened Treg accumulation in the infarct border zone. This was paralleled by a lower proportion of TNF-producing cells among cardiac monocytes and macrophages reflecting a shift away from inflammation and towards wound healing. Taken together, our findings in the pig MI model are the first to show that low-dose CD28-SA infusion is capable of ameliorating tissue inflammation in a species prone to CRS development after high-dose CD28-SA infusion. Moreover, the data highlight that therapeutically shifting the Treg/Tconv balance in MI patients might be feasible in order to enhance cardiac wound healing and prevent adverse remodelling.

immunology↗

bioRxiv2025

Interferon gamma signaling drives cardiac metabolic rewiring

BackgroundIFN-gamma (IFN-{gamma}) signaling influences myocardial inflammation and fibrosis across a wide range of conditions, including ischemic and non-ischemic heart failure (HF). However, the direct effects of IFN-{gamma} on cardiomyocytes remain poorly understood. Here, we developed a novel in vivo model to investigate how IFN-{gamma} impacts myocardial metabolism and function. MethodsMale C57BL/6J mice were injected intravenously with hepatotropic adeno-associated virus (AAV2/8) carrying Ifng and nLuc reporter under the albumin promoter (AAV- Ifng) or empty vector control virus (AAV-ctrl). Cardiac alterations were monitored on day 28 through flow cytometry, bulk RNA sequencing, targeted metabolomics, isolated mitochondrial activity, echocardiography, and in vivo imaging using [18F]fluordeoxyglucose ([18F]FDG) and [18F]fluoro-6-thia-heptadecanoic acid. Additionally, mice lacking IFN-{gamma} receptor expression in cardiomyocytes (Myh6Cre Ifngr1fl/fl) were used to further dissect the cell-intrinsic roles of IFN-{gamma} signaling in cardiomyocyte metabolic reprograming. ResultsAfter confirming liver-specific viral transfection and elevated serum IFN-{gamma} production at physiological levels, we observed cardiac metabolic adaptation and rewiring in animals treated with AAV-Ifng compared to control animals. Myocardial bulk RNA sequencing and gene set enrichment analysis identified an IFN-{gamma} response signature accompanied by marked down-regulations of oxidative phosphorylation and fatty acid oxidation pathways. Functional assessment of isolated cardiac mitochondria showed decreased oxygen consumption, and targeted metabolomics confirmed metabolic shifts toward glycolysis in mice overexpressing IFN-{gamma}. In vivo imaging confirmed increased cardiac glucose uptake following AAV-Ifng treatment. Notably, these metabolic alterations were abrogated in mice with cardiomyocyte-specific deletion of IFN-{gamma} receptors (IFNGR). ConclusionsSystemic IFN-{gamma} induces pronounced metabolic reprogramming in the heart, characterized by increased glucose uptake and reduced oxidative phosphorylation, via direct signaling through cardiomyocyte IFNGR. These alterations mirror those observed in aging and some forms of HF, thereby highlighting that, beyond classical inflammation, this cytokine regulates cardiac metabolism. Novelty and significanceO_ST_ABSWhat is known?C_ST_ABSO_LIImmunological mechanisms can impact myocardial disease progression through complex context-dependent mechanisms. C_LIO_LIIFN-{gamma}, a cytokine primarily secreted by natural killer and T cells, promotes myocardial inflammation and fibrosis in the context of autoimmune myocarditis, pressure-overload-induced heart failure, and Chagas cardiomyopathy. C_LIO_LICytokines exert pleiotropic effects and can influence inflammatory responses through mechanisms involving control of energy metabolism. C_LI What new information does this article contribute?O_LIA novel adeno-associated virus model of systemic IFN-{gamma} elevation allows assessment of cardio-immune-metabolic crosstalk without confounding factors. C_LIO_LIIFN-{gamma} drives cardiac metabolic reprogramming in inflammatory contexts, characterized by enhanced glucose uptake and glycolysis with mitochondrial dysfunction, ultimately altering cardiac metabolic fluxes and function. C_LIO_LIThe IFN-{gamma}-induced cardiac metabolic reprogramming is, at least in part, mediated through direct signaling via receptors on cardiomyocytes. C_LI

immunology↗

bioRxiv2022

Rapid differentiation of regulatory CD4+ T cells in the infarcted myocardium blunts in situ inflammation

BackgroundMyocardial infarction (MI) is a sterile inflammatory condition associated with tissue injury that results in the activation of T helper cell targeting cardiac antigens. However, the differentiation trajectories and in situ activity of heart-specific CD4+T cells activated in the MI context remain poorly understood. MethodsHerein, we combined T-cell receptor transgenic models targeting myocardial protein, single-cell transcriptomics, and functional phenotyping to elucidate how the myosin-specific CD4+ T cells (TCR-M) differentiate in the murine infarcted myocardium and ultimately influence tissue repair. Furthermore, we adoptively transferred heart-specific T-cells that were pre-differentiated in vitro towards pro-inflammatory versus regulatory phenotypic states to dissect how they differentially regulate post-myocardial infarction (MI) inflammation. ResultsFlow cytometry and single-cell transcriptomics findings reveled that transferred TCR-M cells rapidly acquired an induced regulatory phenotype (iTreg) in the infarcted myocardium and blunt local inflammation. Myocardial TCR-M cells differentiated into two main lineages enriched with cell activation and pro-fibrotic transcripts (e.g. Tgfb1) or with suppressor immune checkpoints (e.g. Pdcd1), which we also found in human myocardial tissue. These cells produced high levels of latency-associated peptide (LAP) and inhibited interleukine-17 (IL-17) responses. Notably, TCR-M cells that were pre-differentiated in vitro towards a regulatory phenotype maintained a stable in vivo FOXP3 expression and anti-inflammatory activity when adoptively transferred prior to MI induction. In contrast, TCR-M cells that were pre-differentiated in vitro towards a pro-inflammatory TH17 phenotype were partially converted towards a regulatory phenotype in the injured myocardium and blunted myocardial inflammation. ConclusionsThese findings reveal that the myocardial milieu provides a suitable environment for iTreg differentiation and reveals novels mechanisms by which the healing myocardium shapes local immunological processes.

immunology↗

bioRxiv2021

Diversify your workflow! - An inconvenient suggestion to analyze spike data from intracranial recordings

An important challenge of neuroscience research and future brain machine interfacing is the reliable assignment of spikes to putative neurons. By means of extracellular recordings, researchers try to match different types action potentials with their putative neuronal source and timing. Unfortunately, this procedure is by far not standardized and reliable, leading to many different suggestions and as many differing results. It appears that sharing of data is thus hampered by different processing pipelines in different labs, thus playing along the reproducibility crisis in neurosciences. To systematically shed light on this issue, we present preliminary results of several easy event detection schemes on one data set, meant to illustrate the inconsistencies arising from different processing pipelines already in its initial step. The results indicate that thresholding choices alter findings due to a lack of a ground truth for spike sorting. We suggest to increase reliability in findings by only accepting and further processing events accepted by more than one processing pipeline.

neuroscience↗

bioRxiv2020

Bilateral Intracranial Beta Activity during Forced and Spontaneous Movements in a Hemi-PD Rat Model

Cortico-basal ganglia beta oscillations (13-30Hz) are assumed to be involved in motor impairments in Parkinsons Disease (PD), especially in bradykinesia and rigidity. Various studies have utilized the unilateral 6-OHDA rat PD model to further investigate PD and test novel treatments. However, a detailed behavioral and electrophysiological characterization of the model, including analyses of popular PD treatments such as DBS, has not been documented in the literature. We hence challenged the 6-OHDA rat PD model with a series of experiments (i.e. cylinder test, open field test and rotarod test) aimed at assessing the motor impairments, analyzing the effects of Deep Brain Stimulation (DBS), and identifying under which conditions excessive beta oscillations occur. We found that hemi-PD rats presented an impaired performance in all experiments compared to the sham group, and DBS could improve their overall performance. Across all the experiments and behaviors, the power in the high beta band was observed to be an important biomarker for PD as it showed differences between healthy and lesioned hemispheres and between PD and sham rats. This all shows that the 6-OHDA PD model accurately represents many of the motor and electrophysiological symptoms of PD and makes it a useful tool for the pre-clinical testing of new treatments and further investigations into this disease.

neuroscience↗

bioRxiv2020

Long-term in vivo Monitoring of Gliotic Sheathing of Ultrathin Entropic Coated Brain Microprobes with Fiber-based Optical Coherence Tomography

Microfabricated neuroprosthetic devices have made possible important observations on neuron activity; however, long-term high-fidelity recording performance of these devices has yet to be realized. Tissue-device interactions appear to be a primary source of lost recording performance. The current state of the art for visualizing the tissue response surrounding brain implants in animals is Immunohistochemistry + Confocal Microscopy, which is mainly performed after sacrificing the animal. Monitoring the tissue response as it develops could reveal important features of the response which may inform improvements in electrode design. Optical Coherence Tomography (OCT), an imaging technique commonly used in ophthalmology, has already been adapted for imaging of brain tissue. Here, we use OCT to achieve real-time, in vivo monitoring of the tissue response surrounding chronically implanted neural devices. The employed tissue-response-provoking implants are coated with a plasma-deposited nanofilms, which have been demonstrated as a biocompatible and anti-inflammatory interface for indwelling devices. We evaluate the method by comparing the OCT results to traditional histology qualitatively and quantitatively. The differences in OCT signal across the implantation period between the plasma group and the control reveal that the Parylene-type coating of otherwise rigid brain probes (glass and silicon) does not improve the glial encapsulation in the brain parenchyma.

bioengineering↗

bioRxiv2018

Intravital optoacoustic ultrasound bio-microscopy reveals radiation-induced skull vasculopathy

Angiogenesis is critical in bone development and growth. Dense, large-scale, and multi-layered vascular networks formed by thin-walled sinusoidal vessels perfuse the plate bones and play an important role in bone repair. Yet, the intricate functional morphology of skull microvasculature remains poorly understood as it is difficult to visualize using existing intravital microscopy techniques. Here we introduced an intravital fully-transcranial imaging approach based on hybrid optoacoustic and ultrasound bio-microscopy, allowing for large-scale observations and quantitative analysis of the vascular morphology, angiogenesis, vessel remodeling, and subsurface roughness in murine skulls. Our approach also enabled high-throughput physiological studies to understand radiation-inhibited angiogenesis in the skull bone. We observed previously undocumented sinusoidal vascular networks spanning the entire skullcap, thus opening new vistas for studying the complex interactions between calvarian, pial, and cortical vascular systems.

bioengineering↗

bioRxiv2017

Mud sticks after all: Chronically implanted microelectrodes but not electrical stimulation cause c-fos expression along their trajectory

The goal of CNS implanted devices is to build a stable brain-machine-interface. The brain tissue response to the foreign body limits the functionality and viability of this brain-machine connection. Notably the astrocytic glial scar formation and inflammation with resulting neuronal cell loss is considered to be responsible for the signal deterioration over time. We chronically implanted a polyimide microelectrode in the dorsolateral striatum of rats. First, we analyzed the c-fos immunoreactivity following high frequency stimulation (HFS) of the dorsolateral striatum and second, using GFAP and ED1 immunocytochemistry, the brain tissue response. Acute as well as chronic HFS showed no significant change of neuronal c-fos expression in the dorsolateral striatum and corresponding cortical areas. We found that the sole chronic implantation of a polyimide microelectrode leads to a reaction of the surrounding neurons, i.e. c-fos expression, along the implantation trajectory. We also observed the formation of a glial scar around the microelectrode with a low number of inflammation cells. Histological and statistical analysis of NeuN positive cells showed no kill zone, which accompanied neuronal cell death around the implantation site.

neuroscience↗