Biology subjects

Jacoby, E.

Publications and source records attributed to Jacoby, E..

5 recordsLinked to original sources

bioRxiv2026

Dual Immune Checkpoint and Cytokine Receptor Modulation by an Engineered Human CTLA-4/IL-10 Bispecific Fusion Protein

Bispecific fusion proteins represent a unique strategy for developing precision therapeutics. By linking functional domains from distinct proteins, these biomolecules can engage multiple targets, enhancing both therapeutic efficacy and safety. Unlike bispecific antibodies, low-molecular-weight fusion proteins offer distinct advantages, including reduced immunogenicity and superior tissue penetration due to their relatively compact size and structure. Such a profile is particularly valuable in managing complex inflammatory diseases, where modulating multiple pathways is required to impart an effective anti-inflammatory effect. Among the various regulators of immune signaling, the cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and interleukin-10 (IL-10) play imperative roles in immune suppression through their interactions with CD80/86 and IL-10R, respectively. While Fc-fused CTLA-4 is a clinically approved drug (e.g., Abatacept), the clinical development of IL-10 has been hampered by unpredictable immunostimulatory side effects. Here, we engineered a bispecific fusion protein linking the extracellular domain of CTLA-4 to IL-10. We successfully expressed the protein in E. coli as an N-terminal GST-tagged variant and refolded it from the inclusion bodies. Additionally, we achieved soluble expression of an Fc-tagged variant in mammalian CHO cells. Both origins demonstrated binding to their cognate receptors, CD80 and IL-10R. Finally, the fusion protein demonstrated a T cell-inhibitory effect by reducing Interferon-{gamma} (IFN{gamma}) secretion levels in an in vitro human Virus-Specific T cells (VSTs) model. This innovative protein engineering offers a promising strategy for addressing unmet clinical needs in autoimmune and inflammatory diseases.

bioengineering↗

bioRxiv2026

Sustained exposure to CAR-T cell secretome impairs human Hematopoietic Stem Cell function and is reversible by dual TNFα-IFNγ blockade

Prolonged cytopenias are a frequent complication of chimeric antigen receptor (CAR) T-cell therapies and are associated with increased infection risk and non-relapse mortality. Although inflammatory cytokines released during CAR-T cell activation have been implicated in immune effector cell-associated hematotoxicity (ICAHT), their direct effects on human hematopoietic stem and progenitor cells (HSPCs) function remains incompletely understood. Here, we established a reductionist model of CAR-T-associated hematotoxicity using conditioned media (CM) derived from activated CD19 CAR-T cells. Sustained exposure of human HSPCs to CAR-T-derived inflammatory secretome impaired HSPC expansion and reduced long-term repopulating capacity in xenotransplantation assays. In contrast, short-term exposure did not abrogate HSPC function, indicating that brief inflammatory signals can initiate durable reprogramming events, with functional consequences emerging during subsequent proliferative expansion. Mechanistically, CAR-T CM induced IFN{gamma}- and TNF-responsive transcriptional programs in HSPCs and promoted inflammatory myeloid skewing without evidence of apoptosis-dependent stem cell loss. Combined inhibition of IFN{gamma} and TNF restored HSPC expansion, normalized lineage output, reversed inflammatory transcriptional signatures, and rescued in vivo repopulating capacity without impairing CAR-T cytotoxic activity. These findings demonstrate that CAR-T-derived inflammatory signaling can directly impair human HSC function and identify dual TNF/ IFN{gamma} blockade as a potential strategy to mitigate CAR-T-associated hematotoxicity while preserving antitumor efficacy.

cell biology↗

bioRxiv2026

Engaging Visual Media Shifts Taste-Related Neural Processing: An fMRI Study on Distracted Eating

BackgroundDistracted eating is prevalent in modern environments. While behavioral research consistently shows that distraction attenuates taste perception and increases food intake, the underlying neural mechanisms appear to be more complex. ObjectiveThis functional magnetic resonance imaging (fMRI) study investigated whether naturalistic distraction modulates gustatory processing via sensory suppression or reallocation of neural resources, as observed in more controlled cognitive load paradigms. MethodsThirty-eight healthy participants received sweet and umami taste stimuli of low and high concentration during fMRI scanning. Attentional state was manipulated using short food-related (low-distraction) versus film-related (high-distraction) video clips. After each video, participants rated perceived intensity and pleasantness. Group-level analyses included covariates for sex, body mass index (BMI), and hunger level. ResultsHigh distraction attenuated perceived intensity (p < 0.001, d = -0.28) and pleasantness (p < 0.01, d = -0.21), independent of taste category or concentration. No significant attenuation by distraction was observed in core gustatory regions (insula, orbitofrontal cortex). Instead, distraction increased activation in occipital, thalamic, and cerebellar regions, indicating a redistribution of processing resources toward visual and attentional systems. ConclusionDistraction reduced taste salience without lowering gustatory cortex activity, supporting resource-competition models rather than active sensory suppression. These results reinforce that the impact of distracted eating is behaviorally robust yet neurally subtle, highlighting the need for personalized stimuli and ecologically valid methods to capture real-world eating behavior. The study demonstrates that video-based paradigms work reliably in fMRI and capture how naturalistic distraction alters taste experience.

neuroscience↗

bioRxiv2025

CAR T-cells dysfunction in the central nervous system is mediated by BBB-induced activation-induced cell death

Chimeric Antigen Receptor (CAR) T-cells can eliminate leukemia and lymphoma within the central nervous system (CNS). Following a case with CD19+ relapse in the CNS in the presence of persisting Tisagenlecleucel, we generated several models to study CAR T-cells in the CNS. In both immunocompetent and humanized leukemia-bearing mice, CNS-residing CAR T-cells were shown to have inferior ex-vivo killing capacities compared to CAR T-cells harvested from the spleen or bone marrow of receptive mice. In a mechanistic work-up, we identified two contributing factors: First, CAR T-cells cultured in cerebrospinal fluid showed impaired cytotoxic and proliferative function compared to those in serum or other media. Second, using a unique in vitro human blood-brain barrier (BBB) model composed of endothelial cells and brain pericytes, we demonstrated that CAR T-cells that crossed the BBB exhibited antigen-independent activation and diminished cytolytic capacity. This led to increased activation-induced cell death of the BBB-migrating CAR T-cells and was confirmed to be independent of the CAR target and the costimulatory domain. Our findings add possible mechanisms of CAR T-cell resistance in the CNS, relevant for CNS hematologic and non-hematologic tumors.

cancer biology↗

bioRxiv2023

Improved CAR-T cell activity associated with increased mitochondrial function primed by galactose

CD19 CAR-T cells have led to durable remissions in patients with refractory B-cell malignancies; nevertheless, most patients eventually relapse in the long term. Many interventions aimed at improving current products have been reported, with a subset of them focusing on a direct or indirect link to the metabolic state of the CAR-T cells. We assessed clinical products from an ongoing clinical trial utilizing CD19-28z CAR-T cells from patients with acute lymphoblastic leukemia. CAR-T clinical products leading to a complete response had significantly higher mitochondrial function (by oxygen consumption rate) irrespective of mitochondrial content. Next, we replaced the carbon source of the media from glucose to galactose to impact cellular metabolism. Galactose-containing media increased mitochondrial activity in CAR-T cells, and improved in vitro efficacy, without any consistent phenotypic change in memory profile. Finally, CAR-T cells produced in galactose-based glucose-free media resulted in increased mitochondrial activity. Using an in vivo model of Nalm6 injected mice, galactose-primed CAR-T cells significantly improved leukemia-free survival compared to standard glucose-cultured CAR-T cells. Our results prove the significance of mitochondrial metabolism on CAR-T cell efficacy and suggest a translational pathway to improve clinical products.

immunology↗