bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular mechanism for adenosine regulation of dendritic cells
Authors: Cekic Caglar , Kayhan Merve , Koyas Altay , Akdemir Imran , Savas Ali Can 
Published: January 2023

Abstract
Extracellular adenosine concentrations and expression adenosine receptors are elevated after cell death, injury and inflammation. Antigen presenting cells are one of the primary targets for adenosine to suppress T and NK cell responses. However, molecular mechanisms for adenosine regulation of dendritic cells are poorly understood. Here we show that adenosine receptor stimulation strongly suppresses dendritic cell activation. Adenosine did not affect phosphoactivation of inflammatory signaling pathways such as MAPKs, NF-κB and IRF3. Adenosine receptor signaling increased intracellular cAMP accumulation. Specific cAMP analogs for EPAC and PKA pathways phenocopied the effects of adenosine on dendritic cells especially when used together. Adenosine-mediated suppression of inflammatory/effector cytokines is associated with increased expression of anti-inflammatory c-Fos and NR4A receptors. NR4A and cFos expression were also increased by EPAC and PKA-specific cAMP analogs. Addition of conditioned mediums from dendritic cells prestimulated with stable adenosine analog or PKA and EPAC specific cAMP analogs together inhibited the IFNγ production by activated T cells. Overall our data suggest that adenosine can potentially target both PKA and EPAC pathways and increase the expression of NR4A nuclear orphan receptors and cFos to regulate dendritic cell responses. Our findings have important implications for the development of novel therapies for immune related diseases such as cancer by targeting adenosine receptors.
Tissue-resident liver NK cell responses to MCMV infection
Authors: Dodard Garvin , Erick Timothy , Tata Angela , Brossay Laurent 
Published: January 2023

Abstract
Natural Killer (NK) cells belong to the ILC1 subgroup, which contains conventional NK (cNK) cells, ILC1 and tissue-resident NK (trNK) cells. The murine liver is home to both cNK cells and trNK cells. In this organ, cNK cells and trNK cells can be distinguished based on the expression of the transcription factors Eomes and T-bet. cNK cells are double positive for Eomes and T-bet while trNK cells do not express Eomes. During viral infection, the fate of the activated cNK cell population is relatively well defined. However, the fate of the trNK cell population remains unexplored. Using the MCMV model of infection, we found that a contraction phase precedes a replenishment of the trNK cell subset, which inversely correlates with cNK cell kinetics. We sought to determine whether the trNK cell response was a direct result of MCMV infection or an indirect response to inflammatory milieu. In order to help distinguish between these two possibilities, mice were injected with the iNKT ligand a-galactosylceramide (a-GalCer). We found that treatment with a-GalCer lead to an initial contraction phase of liver trNK cells, indicating that the phenotype observed is independent of MCMV infection. Altogether, these data suggest that liver cNK and trNK cell subsets, which arise from distinct developmental pathways, perform different functions in this organ.
CoCr micro-particles induce a robust type 2 inflammation in knee synovial tissues leading to fibrosis and tissue damage
Authors: Palma Mark J , Mishra Pankaj Kumar , Millman Ariel , Hallab Nadim J , Beebe Kathleen , Benevenia Joseph , Gause William C 
Published: January 2023

Abstract
Cobalt chrome (CoCr) is a major component of orthopaedic implants and a source of metallic wear debris, which may contribute to harmful inflammation and implant failure. In this study we show that the presence of M2 macrophages (CD68+, CD206+, and CD163+) and tissue fibrosis is observed in human peri-prosthetic tissue samples obtained from patients undergoing revision joint arthroplasty. We further examined the molecular mechanisms that drive the sterile inflammatory response to CoCr microparticles in 2 clinically relevant murine articular models. Bilateral intra-articular knee injections of CoCr microparticles were given to C57BL/6 mice and innate immune cell infiltrates were examined in both acute (2 day) and chronic (2 week) models. At the acute time point, CoCr microparticles induce inflammation characterized by an innate type 2 response in synovial tissues with marked increases in neutrophils, eosinophils, and M2 macrophages. This response in articular tissues is dependent on both SYK and BTK signaling as pharmacologic blockade of either pathway abrogates the innate response. Studies in our chronic model demonstrates that repeated exposure of articular tissues to CoCr microparticles triggers an inflammatory response that is characterized by persistent innate cell infiltrates of neutrophils and macrophages, as well as the development of tissue fibrosis. Additionally we show that the innate response and generation of tissue fibrosis in our chronic model is independent of the Caspase-1 inflammasome. Taken together, our findings suggest that CoCr mediates a robust type 2 inflammation that leads to fibrosis and identifies novel potential targets for control of microparticle induced sterile inflammatory responses.
Activation of innate sensing cGAS-STING pathway by Sindbis Virus
Authors: Fung Sin-Yee , Yuen Chun-Kit , Zhu Wuyang , Liang Guodong , Kok Kin-Hang , Chan Chi-Ping , Jin Dong-Yan 
Published: January 2023

Abstract
The cGAS-STING pathway for cytosolic DNA sensing induces type I interferon response during the course of infection with DNA viruses, retroviruses or bacteria. In addition, cGAS detects cytosolic self-DNA, which could be resulted from the reactivation of endogenous retroviral elements or leaked from mitochondria or the nucleus under autoimmune conditions. Cytosolic RNA species can also serve as cGAS ligand in the form of DNA:RNA hybrid. However, the relevance of cGAS-STING pathway to RNA viruses remains unclear. In this study we report the activation of cGAS-STING pathway by Sindbis virus with a positive-sense single-stranded RNA genome. cGAS transcript was induced during Sindbis virus infection and this induction was independent of type I interferon signaling. Loss of either cGAS or STING augmented Sindbis virus replication and prevented viral induction of type I interferons, suggesting a possible role of cGAS-STING signaling pathway in the sensing of RNA viruses. Reconstitution of cGAS and STING in cGAS- and STING-null cells exhibited a protective effect against Sindbis virus infection. Analysis of cytosolic nucleic acid species enriched from Sindbis virus-infected cells indicated the presence of a cGAS ligand. Taken together, our findings reveal an unrecognized role of cGAS-STING pathway for cytosolic DNA sensing in RNA virus infection.
Proteasome inhibition ameliorates experimental Sjögren’s syndrome by suppressing Th17 and B cell responses
Authors: XIAO Fan , LIN Xiang , LU Liwei 
Published: January 2023

Abstract
Primary Sjögren’s syndrome (SS) is a chronic autoimmune disease that mainly affects salivary and lacrimal glands. Currently, there is a lack of effective therapies for SS patients. Increasing evidence indicates that Bortezomib (BTZ), the first proteasome inhibitor approved for treating multiple myeloma, shows beneficial effects in treating lupus. Although clinical findings have revealed increased expression of proteasome in SS patients, it remains unclear whether proteasome inhibition is an effective approach for treating SS. In this study, we evaluated the therapeutic effects of BTZ on experimental SS (ESS) mouse model. Mice were immunized with proteins extracted from salivary glands (SG) to induce ESS and received either vehicle or BTZ treatment. Compared with vehicle-treated counterparts, BTZ-treated mice displayed markedly higher saliva flow rates and ameliorated tissue destructions accompanied with less lymphocytes infiltration in SG. Notably, mice with BTZ treatment showed significantly decreased Th17 but not Th1 responses, and reduced germinal center B cells and plasma cells in peripheral lymphoid organs. In culture, BTZ treatment significantly suppressed Th17 cells differentiation and proliferation. Moreover, adoptively transferred naïve CD4 T cells differentiated into Th17 cells and induced salivary dysfunction in immunized Il17a−/− mice. However, the BTZ-treated recipient mice exhibited suppressed Th17 responses with ameliorated reduction in salivary secretion. Taken together, our findings have shown that BTZ treatment can suppress Th17 and B cell responses and effectively ameliorate ESS development in mice, which may contribute to the validation of BTZ as a promising candidate for treating SS.
GMCSF-MOG abrogates experimental autoimmune encephalomyelitis (EAE) through the induction of MOG-specific regulatory T-cells.
Authors: Moorman Cody D , Curtis Alan Dale , Mannie Mark D 
Published: January 2023

Abstract
FoxP3+ regulatory T-cells (Tregs) play a crucial role in maintaining peripheral tolerance by suppressing auto-reactive T conventional cells (Tcon). Developing a safe therapeutic approach to induce autoantigen-specific Tregs could provide an effective treatment for Multiple Sclerosis. Previous studies have shown that single-chain fusion proteins comprised of murine GM-CSF and the dominant MOG35-55 neuroantigen (NAg), when injected subcutaneously in saline, were potent tolerogens that suppressed experimental autoimmune encephalomyelitis (EAE). To assess the potential involvement of FoxP3+ NAg-specific Tregs in the tolerogenic mechanisms of GMCSF-MOG, we used 2D2-FIG mice that express a transgenic “2D2” T cell repertoire specific for MOG35-55 and express a GFP reporter of FoxP3 expression. The results showed that a single subcutaneous administration of GMCSF-MOG in saline elicited a major population of FoxP3+ Tregs comprised of approximately 10–50% of all circulating 2D2-FIG T cells. This GMCSF-MOG-induced FoxP3+ Treg (GMiTreg) subset appeared within 3 days, was sustained over several weeks, and was associated with a desensitized phenotype within the Tcon compartment including a decrement in circulating transgenic 2D2 T cells, and down-regulation of TCR, CD3, and CD4 on a per cell basis. Covalent linkage between GM-CSF and MOG35-55 was required for the induction of FoxP3+ Tregs and the desensitized phenotype. The tolerogenic activity of GMCSF-MOG was dependent upon CD25+ GMiTregs, because treatment of mice with the anti-CD25 mAb PC61 abrogated GMiTregs and completely reversed vaccine-induced tolerance. This study indicates that immunization with GMCSF-MOG induces antigen-specific Tregs, which inhibit EAE in mice.
Potential Role of Nitrobiolome in Autoimmune Disease: Efficacy of GSNO in EAE.
Authors: Singh Inderjit , Saxena Nishant , Singh Avtar K , Won Jeseong 
Published: January 2023

Abstract
S-nitrosoglutathione (GSNO) is a physiological nitric oxide (NO) carrier molecule important in modulation of NO-signaling. Cellular GSNO homeostasis is maintained by its synthesis by nitric oxide synthase (NOS) as well as its catabolism by GSNO reductase (GSNOR). Treatment of EAE mice (wild-type /WT) with exogenous GSNO or GSNOR inhibitor (N6022) significantly alleviated the clinical disease and associated myelin damage. In addition, more profound alleviation of EAE disease was observed in mice deficient in GSNOR (GSNOR−/−). However, EAE disease was further exacerbated in mice deficient in inducible NOS (iNOS−/−) as compared to WT mice. As expected, the spleen levels of S-nitrosothiol proteins were higher in GSNOR−/− EAE mice but lower in iNOS−/− EAE mice compared to WT EAE mice indicating a negative correlation of spleen GSNO levels with EAE disease severities. Both GSNO and N6022 treated EAE mice had decreased TH17 and increased CD4+/CD25+/FOXP3− Treg differentiations without any significant alterations in other CD4+ T cell subset differentiation. Accordingly, CD4 T+ cells from GSNO or N6022 treated EAE mice produced lower amount of IL-17 and higher amount of IL-10 compared to the cells from untreated WT EAE mice. GSNOR−/− EAE mice also exhibited decreased T 17 and increased CD4+/CD25+/FOXP3− Treg differentiations. On the other hand, iNOS−/− EAE mice exhibited increased TH17, but decreased Tregs differentiations compared to WT EAE mice. In summary, these studies document that nitrobiolome (GSNO) participates in differentiation of autoreactive TH17 and Treg subsets during the course of EAE, and that GSNO as a potential therapeutic drug for multiple sclerosis. Supported in-part by VA(BX002829) and NIH(NS037766).
Vector-Free Engineering of Antigen Presenting Cells for Adoptive Immunotherapies for the Treatment of Cancer
Authors: Myint Melissa , Loughhead Scott , Merino Amy , Tian Lina , Bernstein Howard , Sharei Armon 
Published: January 2023

Abstract
In recent years, ex vivo manipulation of primary cells has shown immense clinical potential with the advent of adoptive T cell therapies. Conventional methods for ex vivo manipulation, however, are not without limitations. They typically rely on the application of electrical fields or exogenous materials such as viral vectors and plasmids, which can increase the potential for cellular toxicity and off-target effects. To overcome such limitations, we have developed an approach using our CellSqueeze Technology that causes temporary membrane disruption as cells are passed through a microfluidic constriction. While the membrane is disrupted, material in the surrounding buffer can diffuse directly into the cytosol. This system has demonstrated efficacy in patient-derived cells, such as stem cells and immune cells, and with a variety of molecules that are difficult to address with alternative methods.
In this work, we describe the use of our vector-free technology to deliver antigens directly to the cytoplasm of antigen-presenting cells (APCs) to drive potent antigen-specific CD8 T cell responses. Conventional methods for eliciting T cell responses with APCs typically rely on cross-presentation, which can be inefficient and require lengthy cultures. Our results show that murine APCs processed with our CellSqueeze Technology can stimulate enhanced antigen-specific T cell responses in vitro and in vivo by at least 3-fold, when compared to responses stimulated by endocytosis controls. Additionally, we show translation of these significant advantages of the CellSqueeze Technology to human APCs, reinforcing the exciting clinical potential of CellSqueeze for adoptive cell therapy.
Peptide-based polymer Toll-Like Receptor 7/8 agonist nanoparticles increase the breadth of anti-tumor CD8 T cell immunity
Authors: Ramirez-Valdez Andrei , Blobel Nicolas J , Ishizuka Andrew S , Pan Jenny , Vodnala Suman , Restifo Nicholas P , Lynn Geoffrey M , Seder Robert A 
Published: January 2023

Abstract
Targeting tumor specific neoantigens through personalized therapeutic vaccination is a promising approach to induce T cell immunity against a variety of tumors. Current vaccine formulations induce CD8 T cell responses against a limited subset of pre-selected neoantigens. We developed a novel vaccine platform to enhance the breadth and magnitude of CD8 T cell responses by chemically coupling synthetic neoantigen peptides to a synthetic polymer containing a toll-like receptor-7/8 agonist that self-assembles into nanoparticles (termed SPP-7/8a). To test the efficiency of the SPP-7/8a platform for inducing antigenic breadth, 173 transcribed non-synonymous single nucleotide variants were identified in a mouse melanoma tumor line (SB-3123). Mice were vaccinated twice with a random set of 96 SB-3123 neoantigens with SPP-7/8a. The binding affinity was predicted for each neoantigen using the Immune Epitope Database and Analysis Resource (IEDB.org) Consensus algorithm, which ranged from the 0.15 to 3.05 percentile. Overall, CD8 T cell responses were detectable at frequencies ranging from 0.1% to 16% of total CD8 T cells against 10 of the 96 neoantigens (~10%). However, by stratifying the 96 neoantigens according to high affinity binders (Consensus &lt; 0.5), 7/14 (50%) induced CD8 T cell responses. These data show that the SPP-7/8a platform efficiently induces CD8 T cell using peptides that have high binding affinity. Ongoing studies will determine whether the increased breadth of CD8 responses following SPP-7/8a vaccination confers protection against established tumors.
Optimization of Gene Editing in Human Primary T Cells with Neon® Transfection System
Authors: Chang Sean , Andronikou Nektaria , de Mollerat du Jeu Xavier 
Published: January 2023

Abstract
The successful cases of autologous CAR-T cell therapy in leukemia have highlighted the promising future of cell therapy. However, most studies in this field have been using viruses to engineer T cells, which lead to safety concerns. On the other hand, there has been a growing interest in developing allogeneic immune cell therapies to tackle the challenges of production scale up. Hence, there is a need for a better delivery system to maximize gene editing efficiency in immune cells.
Neon® Transfection System has been widely used in a variety of cell models and especially hard-to-transfect cells, such as primary cells and stem cells. Here we demonstrate how Neon® is fully capable of supporting all spectrums of research in cell therapy, including RNA delivery, DNA delivery, and CRISPR/Cas9 gene editing. Human primary T cells from four different donors were isolated from individual fresh Leukapheresis Pak, followed by activation/expansion in OpTmizer media with 2% human serum and CD3/CD28 Dynabeads. The optimal delivery time was between 3–5 days after activation. The transfection efficiency can be reached over 95% for RNA (EGFP mRNA) and 85% for DNA (pcDNAEF1a-emGFP) with high viability using certain Neon® programs. For CRISPR/Cas9 gene editing, more than 30% knock-in efficiency was observed while cleavage efficiency at HPRT locus can be reached more than 70%.
The promising results of gene delivery and editing in primary T cells from Neon® could facilitate the progress of immunotherapy. Meanwhile, we have been dedicating in developing a next generation cGMP-certified large-scale electroporation system, which enable researchers to seamlessly scale-up from Neon®.
Detection and tracking of public, broadly neutralizing antibodies against group 1 and group 2 Influenza A viruses in healthy donors and post-seasonal flu vaccine.
Authors: Gittelman Rachel M , Yusko Erik , Sanders Catherine M , Vignali Marissa , McDermott Adrian , Robins Harlan 
Published: January 2023

Abstract
Influenza A viruses form two diverse clades that co-circulate and cause seasonal infections each year. This diversity has made it challenging to develop a universal flu vaccine that effectively targets epitopes across both clades. One approach to developing a universal flu vaccine is to identify broadly neutralizing antibodies that can be reproducibly elicited across individuals by novel immunogens or immunogens specifically designed to enhance these sequences. These public antibodies may represent common and accessible routes to protection against influenza in the general population. Next generation sequencing can be used to monitor vaccine trials for the presence of these sequences. In this work, we leverage Adaptive Biotechnologies’ IGH sequence database and immunoSEQ® assay to track and characterize a set of recently identified public IGH motifs that are capable of neutralizing group 1 and group 2 flu viruses1. We first characterized the incidence and basal frequency of these motifs in blood from 3 healthy individuals, as well as bone marrow from 97 patients following immune reconstitution. Next, we monitored clonal expansion and contraction of these motifs in 15 healthy subjects immediately prior to, and at days 3, 7, 14, 28, and 120 post seasonal flu vaccination. Finally, we placed these motifs within the broader context of immune response following seasonal flu vaccination through additional deep sequencing of the IGH and TCRB loci in these subjects. These data show that immunoSequencing can be used as a highly quantitative and sensitive tool to track the production and maintenance of specific antibody sequences.
Adapting the innate immune system to develop long-lived vaccines to bacterial pathogens
Authors: Super Michael , Doherty Edward , Cartwright Mark , Stafford Alexander , Ali Omar , White Des , Graveline Amanda , Urena Frank , MacDonald Chloe , Dinis Alexandre , Duffy Shannon , Duckless Benjamin , Seiler Benjamin , Ingber Donald , Mooney David 
Published: January 2023

Abstract
There is an urgent need to develop vaccines against bacteria due to the rise of Multi-drug resistant (MDR & XDR) organisms. To date, it has been difficult to produce protective vaccines against bacterial pathogens; there is a danger of outgrowth of fast growing attenuated bacterial strains while polysaccharide cell walls are poorly immunogenic and subunit vaccines are ineffective, generating TI (T-independent) immune responses, characterized by low affinity, short lived, non-class switched IgM antibodies.
We are developing technologies for generating vaccines in mice against multiple pathogen species, including bacterial; MDR E. coli, MRSA, MTb LAM, Viral; HIV gp120, parasitic; P. falciparum and fungal; C. albicans. We capture pathogens using FcMBL Opsonin technology (which binds more than 90 different pathogen species), and present the killed pathogens in an immune modulating biomaterial system. This lyophilized product provides long-lasting protection with a single dose through a novel self-boosting mechanism of action.
Our vaccines can protect mice against MDR strains of E. coli which are lethal within 12 hours. We have raised antibodies with titers which are sustained beyond 90 days (to date) with a single vaccination (the biomaterial system has demonstrated titers beyond 1 year in other indications). Using this opsonin and immune modulating technology, E. coli can be captured from the blood and tissues of one animal and used to vaccinate other animals.
Investigating the roles of TNFα and TGF-β1 in Ly-6A-mediated CD4+ T cell death
Authors: Patel Akshay G , Bamezai Anil K 
Published: January 2023

Abstract
Ly-6A, a member of the Ly-6 family of proteins, negatively regulates the antigen receptor-driven clonal expansion of CD4+ T cells by an unknown mechanism. Ly-6A is a 12–14 kDa protein that lacks transmembrane and cytoplasmic domains and is anchored to the cell membrane with a glycosylphosphatidylinositol (GPI) anchor. On immune cells, this cell surface protein also has adhesion and signaling properties. To explore the mechanism of Ly-6A-mediated CD4+ T cell growth inhibition, we have used immortalized CD4+T cell lines as in vitro experimental model. Engaging Ly-6A caused growth inhibition, apoptosis, and cytokine secretion in culture. Ly-6A activation also upregulated p27kip1, a cell cycle inhibitory protein. Additionally, we observed mitochondrial destabilization as gauged by the release of cytochrome C in the cytoplasm, and detection of active Caspases 3 and 9. The simultaneous observance of cytokine transcription and cell death appeared enigmatic. To examine this issue, we hypothesized that CD4+ T cells generate death and growth inhibitory factors after engaging the Ly-6A protein with a monoclonal antibody. The cytokines TNFα and TGF-β1, growth inhibitory and pro-apoptotic molecules, respectively, were examined. We observed that engaging Ly-6A results in growth inhibition and production of TNFα at higher concentrations than what is generated by engaging the antigen receptor. Importantly, blocking antibodies against TNFα decreased cell death in a concentration-dependent manner as evaluated by flow cytometry. Levels of total and free active forms of TGF-β1 were significantly upregulated by CD4+ T cells to stimulation through the antigen receptor complex, but no changes in secretion of TGF-β1 were found after engaging Ly-6A.