bluuebunny/crossref_metadata_2025_split huggingface dataset:
Artificial Targets: a versatile cell-free platform to characterize CAR T cell function in vitro
Authors: Evaristo Cesar , Wang Xueting , Tokarew Nicholas , Borgelt Nadine , Siemer Ramona , Casonato Melo Cristiane , Langer Christian , Kasampalidis Ioannis , Ogusuku Isabella , Cathomen Toni , Gessner Isabel , Dose Christian , Fauerbach Jonathan , Richter Anne 
Published: November 2024

Abstract
Chimeric antigen receptor (CAR) T cells have proven efficacy against a number of cancers, but further research is needed to extend their therapeutic potential. Functional characterization of CAR T cells plays is central across fundamental research and therapeutic development, with increasing needs for standardization. Typically, effector functions are assessed following co-culture with target-expressing cell lines. This poses several limitations, including high variability, allogeneic-induced background, and work and time intensive workflows. To overcome these limitations, we developed a synthetic bead-based platform (“Artificial Targets”, or "AT") to characterize CAR T cell function in vitro. AT could specifically induce CAR T cell activation, and multiple effector functions of CAR T cells, mimicking the response triggered by target cell lines. These include cytotoxic activity, expression and secretion of cytokines, and cell proliferation. Finally, AT demonstrated flexibility to engage multiple co-stimulatory molecules to enhance CAR T cell function, representing a powerful tool for modulating CAR T cell responses. Collectively, our results show that ATs can specifically activate CAR T cells for essential effector functions that could significantly advance standardization of functional assessment of CAR T cells, from early development to clinical applications.
Affinity and formulation of a phenyl glycolipid for stimulation of invariant natural killer T cells to inhibit tumor growth
Authors: Hung Jung-Tung , Chiou Shih-Pin , Yu Alice 
Published: November 2024

Abstract
α-galactosylceramide (α-GalCer) can activate invariant natural killer T (iNKT) cells to modulate immune system. We have synthesized and tested many α-GalCer analogs and found the most potent iNKT-stimulating agent C34, which has two phenyl rings on the acyl chain. The complex of C34 and CD1d showed the highest binding affinity to the T cell receptor of iNKT cells. Moreover, C34 can induce strong Th1-biased cytokines and exhibit potent anticancer efficacy in tumor animal models. However, poor water solubility of C34 limits its clinical application. In this study, we incorporated C34 into PEGylated lipid nanocarriers (PLN), which improves the water solubility of C34. The PLN-C34 shortened the activation time of iNKT cells and maintained C34's ability to activate iNKT cells to secrete cytokines in vitro. Furthermore, PLN-C34 showed similar anticancer efficacy as C34 dissolved in dimethyl sulfoxide to prolong the survival of mice bearing with TC1 or B16F10 tumor. In general, the PLN-C34 preserved the ability to activate iNKT cells and inhibited tumor growth, suggesting that PLN-C34 is a promising candidate for cancer therapy in clinical settings.
Loss of Trex1 in intestinal epithelial cells prevents enteric viral infections
Authors: Zhang Li , Song Jaewon , Gheshm Newsha , Orvedahl Anthony , Lee Sanghyun 
Published: November 2024

Abstract
The host intestinal microbiota comprises bacteria, viruses, fungi, multicellular parasites, and archaea, which directly impact host’s health. Norovirus and rotavirus are agents of acute infectious gastroenteritis with symptoms of vomiting and watery diarrhea. These viruses are restrained by interferons, especially IFN-α and IFN-λ. The Tree-prime Repair Exonuclease 1 (Trex1) is a nonprocessive 3’→5’ exonuclease that degrades cytosolic nucleic acids. Mutations in Trex1 are associated with a broad spectrum of inflammatory and autoimmune diseases. Failure to degrade cytosolic DNA activates the cGAS-STING DNA sensing pathway and ultimately drives chronic type I and III interferon signaling pathway activation. We found persistent murine norovirus (MNoV) and rotavirus infections were prevented in the Trex1-/- mice. However, Trex1-/-cGAS-/- and Trex1-/-Ifnlr1-/- lost the protection against enteric viral infection seen in Trex1-/- mice. Moreover, tissues from the Trex1-/- mice exhibited a robust interferon stimulated genes (ISG) induction in response to MNoV infection compared to their WT controls. The ISG levels were dampened in both Trex1-/-cGAS-/- and Trex1-/-Ifnlr1-/- mice. Furthermore, Trex1 deficiency in gut epithelial cells is sufficient to suppress MNoV infection. In conclusion, Trex1, as a gatekeeper enzyme of the cGAS-STING pathways, has been demonstrated to be a therapeutic target to restrict enterovirus infection.
102
Authors: Zeleniak Ann , Martinez-Martin Nadia , Laddha Saurabh Vishnu , Kurtagic Elma , Greving Carrie , Hammaker Deepa , Dudas Paul , Perrigoue Jacqueline 
Published: November 2024

Abstract
Inflammation-associated fibroblasts (IAFs) are a cell population that emerges in tissue inflammation. In inflammatory bowel diseases (IBD), IAFs correlate with disease severity and are enriched in colon tissue of patients refractory to anti-TNF therapy. IAFs can amplify tissue inflammation and mediate pro-fibrotic matrix remodeling and therefore represent a novel cellular target for treating inflammatory diseases like IBD. To explore the cellular and molecular pathways that mediate transition to an IAF-like phenotype, we developed a co-culture of human PBMCs and primary colonic fibroblasts to study stromal-immune crosstalk in promoting IAFs. Culturing primed PBMCs with fibroblasts in direct contact drives fibroblast transition to an IAF phenotype through upregulation of inflammatory mediators, evidenced by modulation of gene expression and secreted protein profiles. Single-cell RNAseq revealed IAF development is influenced by PBMC priming conditions. Exposure to LPS-primed PBMCs promoted a matrix-remodeling IAF phenotype while PHA-primed PBMCs promoted an immunomodulatory IAF. IAF activation can be dampened by inhibitors, including TNF and JAK inhibitors. This co-culture system can therefore be used to define mechanisms mediating IAF transition and identify IAF states that can be tailored to accentuate pathways for focused interrogation. We aim to leverage this system to identify novel targets and test prospective therapies to modulate IAF transition and downstream activity.
Single chain MHC-I-β2 microglobulin-peptide trimer constructs identify CD8 T cells recognizing low-affinity MHC-I-binding neoepitopes
Authors: Maji Stabonia , Chawla Arshmeet , Singhaviranon Summit , Dempsey Joesph , Chou Steven , Srivastava Pramod 
Published: November 2024

Abstract
Cancer-specific neoepitopes (mutated peptides in tumor cells presented by MHC-I) are under intense scrutiny to understand the anti-tumor CD8 T cell response. Only a small minority of such potential neoepitopes actually elicit tumor control in vivo. A majority of the neoepitopes, that do mediate tumor control in vivo, have very low affinity for MHC-I, ranging from IC50 values between 500 nM and 50,000 nM (Duan et al., 2014; Brennick and George et al., 2021; Ebrahimi-Nik et al., 2021). This contrasts with the strong MHCI-binding (IC50 &lt;100 nM) epitopes of viruses and model antigens (Peters et al., 2020). As expected, we have found it impossible to create MHC-I-tetramers for low-affinity tumor-controlling neoepitopes. Here, we have genetically created single chain trimers that contain MHC-I alleles, β2 microglobulin as well as the low-affinity peptide, for each of the nine low-affinity neoepitopes identified previously in our laboratory (Brennick and George et al., 2021). Such trimers have been purified and characterized and have been used to identify, for the first time, CD8 T cells that recognize low-affinity cancer neoepitopes presented by tumor MHC-I molecules. The characteristics (abundance, amplification, specificity etc) of these low-affinity MHC-I binding neoepitopes shall be presented.
β-adrenergic signaling regulates erythropoiesis and through it; cancer immunity
Authors: Chawla Arshmeet , Hagymasi Adam , Geyer Rory , Mandoiu Ion , Peterson Kenneth , Scanlon Vanessa , Srivastava Pramod , Nevin James 
Published: November 2024

Abstract
The intricate interplay between the nervous and immune systems significantly influences the fate of immune cells in homeostasis as well as under pathological conditions such as cancer. Notably, adrenergic signals from the sympathetic nervous system have been identified as regulators of hematopoiesis. Disruption of sympathetic signaling has been linked to mal-differentiation of myeloid and lymphoid lineages leading to the emergence of Tregs, myeloid-derived suppressor cells, and the exhaustion of CD8+ T cells. We show here that disrupted sympathetic signaling also leads to mal-differentiation of erythroid lineage via the emergence of erythroid precursor cells (EPCs), which, apart from their role in maintenance of the red blood cell pool, contribute significantly to suppression of the anti-tumor CD8+ T cell response.
Specifically, we show here that EPCs are highly regulated by β-adrenergic signaling in naïve as well as tumor-bearing mice. We observe that EPCs actively promote tumor growth in vivo and influence the fate of tumor infiltrating CD8+ T cells by suppressing their proliferation and effector functions. We have reported several aspects of the mechanisms involved in the β-adrenergic regulation of erythropoiesis including the specific receptors involved, the signaling pathway and the kinetics of the effect. This discovery unveils a novel dimension of neuroimmune communication and offers insights that could catalyze the development of new therapeutic approaches to cancer.
Secondary hemophagocytic lymphohistiocytosis with associated central pontine myelinolysis
Authors: Philip Rachel , Alam Mariam 
Published: November 2024

Abstract
A 34-year-old previously healthy male presented to the emergency department with an altered mental state and unexplained intermittent fevers persisting for three weeks. Initially, his symptoms mimicked an ear infection, for which he received a prescription for Augmentin. As his condition deteriorated, additional symptoms emerged, including bilateral ear pain, fever, and diffuse headaches. Initial evaluations revealed acute hepatic encephalopathy, mastoiditis, and pancytopenia. Further tests uncovered hepatosplenomegaly, lymphadenopathy, and reactivated EBV. The patient met six of the eight HLH criteria, including fever above 38.3°C, cytopenia, elevated triglyceride and ferritin levels, and a bone marrow biopsy confirming hemophagocytosis. The treatment plan followed the HLH-2004 protocol, involving an eight-week induction therapy using etoposide, dexamethasone, and cyclosporine. Monitoring of disease-specific markers such as ferritin and IL-2R showed improvement. One month later, the patient experienced a throbbing headache, nausea, photophobia, and vision changes. MRI showed abnormal signals consistent with central pontine myelinolysis. Lumbar puncture findings included pleocytosis, elevated total protein, and a substantial EBV viral load, indicative of progressive CNS-HLH with persistent EBV reactivation. The patient is currently undergoing salvage therapy for EBV reactivation, with a poor prognosis for CNS involvement and allogeneic transplant preparations.
Dendritic cell CTLA-4+ extracellular vesicles as regulators of T cell polarization
Authors: Bisht Nalini 
Published: November 2024

Abstract
Dendritic cells (DC) are professional antigen-presenting cells that prime de novo T cell responses. DC release extracellular vesicles (EV) to communicate environmental context to other cells of the immune system. Our published data demonstrate that CTLA-4 is expressed on the outer surface of DC-secreted EV. DC-released CTLA-4+ EV are internalized by bystander DC and result in downregulation of costimulatory B7 expression on these bystander DC. TH2 polarization of DC results in enhanced secretion of CTLA-4+ EV as compared to TH1 polarized DC which abrogate secretion of CTLA-4+ EV almost entirely. Furthermore, knockdown of CTLA-4 in DC resulted in upregulated CD8+ T-cell proliferation and enhanced antitumor immunity. Here we report additional characterization of how DC-derived EV modulate adaptive immunity. Co-culture of CTLA-4neg EV with naïve T cells induced significant T cell blast formation and enhanced effector functionality of T cells in the form of upregulated granzyme B and IFN-g in comparison to naïve T-cells cultured with CTLA-4+ EV, indicative of a TH1 response. Additionally, CTLA-4neg EV downregulated Treg proliferation compared to CTLA-4+ EV. These data support the hypothesis that DC CTLA-4+ EV are important effector vehicles through which DC propagate TH polarization signals. Future investigations will determine the therapeutic application of the EV-stimulated T cells, and the corresponding transcriptional changes induced.
Characterization of immune population of model murine of muscular dystrophy.
Authors: Romero Hector , García-Pérez Blanca Estela , Vélez-Caballero Sara H , Cano-Martínez Luis Javier , Martínez-Damas Mirna G , Castillo-Cruz Juan , Coral-Vázquez Ramón M , Baltierra-Uribe Shantal Lizbeth 
Published: November 2024

Abstract
Muscular dystrophy comprises a group of genetic diseases leading to progressive weakness and muscle mass loss due to protein deficiency, such as delta-sarcoglycan protein involved in stabilization of the sarcolemma and in signal transduction. Dystrophic muscles exhibit a predominant inflammatory environment characterized by cell infiltrate capable of producing immunostimulatory signals. While it has been reported that myeloid cell infiltrate may contribute to the pathogenesis of chronic myopathies, the specific role of delta-sarcoglycan protein deficiency in the overall immune response profile has not been previously assessed. In this study, we used flow cytometry to evaluate B and T cell populations, macrophages, monocytes, and neutrophils in the spleens of delta-sarcoglycan null and wildtype mice. We observed significant differences only in the F4/80 macrophage population. These findings suggest that delta-sarcoglycan deficiency may play a role in the differentiation of immune cells, particularly macrophages.
An early burst of IL-2 production by CD8 T cells biases their developmental fate decisions
Authors: Ingram Jennifer , Kahan Shannon , Weaver Casey , Harrington Laurie , Zajac Allan 
Published: November 2024

Abstract
The differentiation of CD8 T cell responses is guided by a combination of antigenic, costimulatory, and cytokine signals. Here we show that naïve CD8 T cells undergo a heterogeneous burst of cytokine synthesis within hours following activation. This divergent early response develops with every specificity of CD8 T cells we have checked, manifests in vivo and in vitro, and occurs even prior to the first cell division. Using cytokine reporter systems, we have demonstrated that these early functional differences are consequential for establishing fate biases and directing the gain of effector and memory T cell traits. Furthermore, we have discovered that the initial cell intrinsic synthesis of IL-2 attenuates IL-2-dependent STAT5 signaling. The bifurcation between the abilities of IL-2-producing and non-producing subsets to elaborate STAT5 signaling are consistent with a model in which recently activated CD8 T cells that manufacture IL-2 become less permissive to cytokine-driven differentiation signals, thereby preserving their “stem-like” properties and allowing them to serve as precursors for memory T cells as well as future effectors. By contrast, the non-IL-2-producing contingent remains receptive to extrinsic IL-2 signals allowing a more terminally-differentiated series of effector CD8 T cells, that are legitimized by cytokine signals, to rapidly form in order to facilitate clearance of the antigenic insult.
Metagenomic investigation of intestinal bacterial and parasitic infections in urban canines in Seoul, Korea
Authors: Kim Ju Yeong 
Published: November 2024

Abstract
Approximately 25% of South Koreans own pets, predominantly dogs. Despite this, there's limited research on the fecal organisms of these dogs and the identification of zoonotic pathogens. This study employs metabarcoding analysis to explore these bacterial and parasitic infections in dog feces in Seoul. Fecal samples were collected from 16 pet and 25 stray dogs in the Mapo district of Seoul. Next-generation sequencing was utilized, employing 16S and 18S rRNA amplicon sequencing to identify prokaryotic and eukaryotic pathogens. Significant differences in bacterial and parasitic composition were found between stray and pet dogs, and putative prokaryotic and eukaryotic pathogens were identified, including Helicobacter, Campylobacter, Giardia, Pentatrichomonas, and Cystoisospora. Interestingly, Campylobacter, Giardia, and Pentatrichomonas were found to be significantly more prevalent in stray dogs than in pet dogs. Additionally, subsequent PCR analysis confirmed that some of these pathogens were strains capable of infecting humans. This study offers valuable insights into the zoonotic risks associated with dog populations residing in diverse environments. By identifying and characterizing potential pathogens in dog feces, this study highlights the need for further research into the immunological mechanisms that enable specific parasites to infect both humans and animals simultaneously.
Immune-modifying cold plasma for treatment of periprosthetic orthopedic infection
Authors: Smith Carly , Watkins Amanda , Melvage Autumn , Lucas Abigail , Schaer Thomas , Freeman Theresa 
Published: November 2024

Abstract
Non-antibiotic treatment options are needed to combat biofilm formation in clinical settings, and cold plasma, an energized gas containing a cocktail of reactive oxygen and nitrogen species, may be such a strategy. In vitro, cold plasma is antimicrobial through direct bacterial killing and modulation of virulence factors. Cold plasma treatment may also be immunostimulatory by enhancement of innate immune effector responses.
We evaluated the efficacy of cold plasma for treating periprosthetic infection in an in vivo rat model. Infection mitigation following cold plasma treatment was assessed by measurement of colony forming units (CFU) of infected rats. Immune modulation was also determined using qPCR, Westerns, IHC, RNA sequencing, and histology.
While we observed no reduction in CFUs, RNAseq revealed cold plasma treatment significantly enriched genes associated with neutrophil activation, chemotaxis, and inflammasome activation. IL-1β was increased at the site of infection, coordinating with increased myeloperoxidase protein and positive cells. Draining lymph nodes in plasma-treated animals were markedly smaller than untreated animals, which had significant lymphadenomegaly.
Cold plasma treatment leads to enhanced innate immune cell recruitment and activation at the site of infection. Additional studies will determine if cold plasma treatment in combination with standard clinical antibiotic therapy will improve bacterial clearance following revision surgery.
A Pro-Drug Strategy Enhances Butyrate Bioavailability and Therapeutic Efficacy In Atherosclerosis
Authors: Beckman Taryn , Volpatti Lisa , Cao Shijie , Slezak Anna , Reda Joe , Budina Erica , De Matos Salvador , DeLeon Orlando , Borjas Gustavo , Chang Eugene , Hubbell Jeffrey 
Published: November 2024

Abstract
Butyrate is a gut microbiota derived short chain fatty acid with pleiotropic positive effects on inflammation and metabolism. In atherosclerosis, sodium butyrate has been shown to significantly reduce atherosclerotic lesions, rectify routine metabolic parameters such as LDL-C in the serum, and reduce systemic inflammation. However, its foul odor, low absorption, and fast metabolism limit its therapeutic effectiveness. We engineered a derivate of butyrate attached to the amino acid serine (SerBut) to mask the taste and odor and co-opt amino acid transporters in the gut to increase its bioavailability. In Vitro, SerBut maintains the anti-inflammatory, NFkB suppressing, capacity of sodium butyrate while allowing higher dosing without inducing cytotoxicity. SerBut can be stably, easily, and odorlessly administered in drinking water at higher doses than previously possible. Over 6 weeks of HFD in concurrence with SerBut water, SerBut reduces LDL-C in the serum, plaque accumulation in the aortic root, and monocytes in the aorta of an apolipoprotein-E-/- murine model of atherosclerosis. Bioinformatic analysis of fecal 16S rRNA sequencing reveals Seryl modification of butyrate to be associated with less plaque in the aortic root of mice, while general butyrate treatment was not. SerBut reduces enzymes associated with liver damage and has a systemic tolerogenic effect. SerBut is a promising therapeutic that lowers lipids and lipid driven inflammation without inducing liver toxicity.
Development of conditionally active IL-10 INDUKINE TM molecules for the treatment of inflammatory bowel disease
Authors: Sullivan Jenna , Aderhold Pamela , Brodkin Heather , Nagy-Domonkos Celesztina , Economides Kyriakos , Hicklin Daniel , Lewis Yuka , Magness Leigh , Siedel-Dugan Cynthia , Steuert Zoe , Stieglitz Jessica , Winston William , Salmerson Andres 
Published: November 2024

Abstract
Inflammatory bowel disease (IBD) encompasses two chronic inflammatory diseases of the gastrointestinal tract, Ulcerative Colitis (UC) and Crohn’s disease. Although great strides have been made in the treatment of IBD, subsets of patients do not respond or lose responsiveness to therapy, underlining a need for new treatments. Interleukin 10 (IL-10) is a immunoregulatory cytokine, known to suppress and inhibit inflammatory responses of a variety of immune cells including monocytes, macrophages, and T cells. Polymorphisms associated with reduced expression of IL-10 and IL-10R have been linked with IBD. Clinical trials using recombinant IL-10 in IBD have been unsuccessful, highlighting a need for further engineering to utilize IL-10 as a therapeutic agent. We have developed IL-10 INDUKINE molecules that are designed as prodrugs containing human IL-10, an inactivation domain and a half-life extension domain tethered together by protease-sensitive linkers. IL-10 INDUKINE molecules are peripherally inactive but due to dysregulation of the protease milieu in the inflamed colon, the linkers are cleaved, and IL-10 is released locally. We have identified proprietary linkers which are cleaved by UC and Crohn’s human colon samples. In a mouse model of colitis, IL-10 INDUKINE molecules prevent weight loss, intestinal damage and inhibit inflammatory cytokine production within the colon. Together these data support the potential of an IL-10 INDUKINE molecule for the treatment of IBD.