bluuebunny/crossref_metadata_2025_split huggingface dataset:
Infection with Porcine Reproductive and Respiratory Syndrome virus (PRRSv) impacts immune cell metabolism without evidence of cell-mediated immune response.
Authors: Loving Crystal L , Olson Zahra F , Schweer Wes P , Gabler Nicholas K 
Published: January 2023

Abstract
Specific metabolic pathways affect immune cell differentiation and function, and the metabolic state of leukocytes may provide a signature associated with a particular condition or treatment. Health challenge in food production animals may warrant dietary changes to meet metabolic demand of the immune system. Porcine reproductive and respiratory syndrome virus (PRRSV) infection negatively impacts pig performance, and the PRRSV-specific immune response is inefficient at providing protection. Given the poor cell-mediated response induced by infection, we hypothesized that the metabolic profile of peripheral blood mononuclear cells (PBMC) from PRRSV-infected pigs would not show a shift to glycolysis, which is a hallmark of T effector cells. Using an Extracellular Flux Analyzer (Seahorse Biosciences) we interrogated the glycolytic and mitochondrial function of PBMC following infection of pigs with PRRSV, and PRRSV-specific cell-mediated immunity. Basal glycolysis of PBMC was not significantly impacted by PRRSV infection, but contrary to our hypothesis, glycolytic capacity and reserve were significantly increased in PBMC from PRRSV-infected pigs over non-infected pigs. The effect of PRRSV infection on PBMC mitochondrial respiration was minimal, with a slight increase in spare respiratory capacity detected. Despite the increased glycolytic capacity of PBMC, there was not a detectable PRRSV-specific cell-mediated immune response. Thus, while there was metabolic signature of lymphocyte activation, it was not associated with a functional response.
Diclazuril treatment ineffective at preventing Equine Protozoal Myeloencephalitis relapse in established mouse model.
Authors: Wagner Alayna , Kasmark Leah , Witonsky Sharon , Lindsay David , Leeth Caroline McPhee 
Published: January 2023

Abstract
The debilitating and potentially fatal neurologic disease, Equine Protozoal Myoencephalitis (EPM), is one of the most common neurologic diseases seen in the equine population of the United States. Disease develops as a result of unintentional ingestion of the pathogenic parasite, Sarcocystis neurona. Treatment options consist of the anticoccidial drug Diclazuril and numerous other antiprotozoal drugs. Weeks to months after cessation of antiprotozoal drug treatment and corresponding neurologic improvement, horses may present with clinical disease symptoms again. Little is known whether this reoccurrence of symptoms is from relapse or reinfection. We sought to understand if relapse was possible following appropriate treatment with diclazuril, a commonly used medication for the treatment of EPM. Using a mouse model of equine EPM, we subjected infected mice to treatment with or without diclazuril for 30 and 60 days. All untreated mice developed neurologic symptoms consistent with S. neurona infection within 30 days post infection. All diclazuril treated mice developed neurologic symptoms cessation of treatment. Cerebellum samples were examined for lesions characteristic of those associated with S. neurona infection and immunohistochemically for presences S. neurona. Sera immunoglobulin levels were analyzed to evaluate treatment’s effect on humoral immunity. In conclusion horses treated appropriately with 60 days of diclazuril may be at risk for relapse and more effective treatment options should be explored.
Kinetic Analysis of ZAP70 Phosphorylation in Human T cells: p-Y292, p-Y319, and p-Y493
Authors: Soper David M 
Published: January 2023

Abstract
The 70-kDa zeta chain-associated protein (ZAP-70) is a Syk family protein tyrosine kinase normally expressed in T and NK cells. Patients deficient in ZAP-70 have no functional T cells in the periphery, are prone to repeated infections, and suffer from ZAP-70-related severe combined immunodeficiency (SCID). In some cases of chronic lymphocytic leukemia (CLL), ZAP-70 is aberrantly expressed in B cells and used a prognostic marker to monitor disease progression and therapeutic intervention. In normal T cells, ZAP-70 is required for thymic development, T cell receptor (TCR) mediated signaling, and T cell activation. ZAP-70 consists of two tandem Src homology 2 (SH2) domains, two linker regions, and a kinase domain. Following TCR engagement, several tyrosine residues within ZAP-70 are phosphorylated and serve to both positively and negatively regulate kinase activity. A kinetic analysis was performed using human peripheral blood mononuclear cells (PMBCs) stimulated with hydrogen peroxide (H2O2), anti-CD3/CD28, anti-CD3/PD-1, or anti-CD3/TIGIT. Using in-house generated clones, T cells were analyzed for three distinct ZAP-70 phospho-epitopes: p-Y292, p-Y319, and p-Y493.
Coalescence of Nanoscale Cytoplasmic Signalosomes Contributes to T Cell Receptor Signaling to NF-κB
Authors: Traver Maria K , Campanello Leonard , Paul Suman , Shroff Hari , Losert Wolfgang , Schaefer Brian C 
Published: January 2023

Abstract
T cell receptor (TCR) activation of the transcription factor NF-κB is a crucial determinant of effector T lymphocyte function. The complex regulatory network surrounding this pathway remains poorly understood, particularly at time periods further removed from initial TCR triggering. We have previously demonstrated that following activation of the TCR, the proteins p62, Bcl10, and Malt1 rapidly combine to form a cytoplasmic filamentous signalosome called POLKADOTS, which recruits further signaling proteins and initiates the terminal steps in activation of NF-κB. Here, we examine the fate of POLKADOTS signalosomes following initial activation of NF-κB. We demonstrate via super-resolution and confocal microscopy techniques that POLKADOTS filaments converge on the microtubule-organizing center via microtubule transport, aggregating in a peri-nuclear aggresomal structure. Aggresomes are poorly understood structures thought to be depots of misfolded protein destined for degradation; however, our data suggest that the aggresomal accumulations of POLKADOTS continue to promote T cell activation. We show that the formation and maintenance of this aggresome corresponds with a secondary increase in NF-κB translocation. Additionally, the aggresomal structure prolongs phosphorylation of IKK and promotes the proteolytic activity of Malt1. Together, these results demonstrate that TCR signaling to NF-κB directs the orchestrated assembly, transport, and stable coalescence of nanoscale cytoplasmic signaling complexes. More broadly, our findings provide evidence that aggresomes can serve as stable platforms of signal transduction, thereby proposing a new role for these little understood structures.
Molecular mechanisms for adenosine regulation of helper T cell activation
Authors: Cekic Caglar , Akdemir Imran , Koyas Altay , Kayhan Merve , Savas Ali Can 
Published: January 2023

Abstract
Polarization of T helper subsets into different functional phenotypes strongly affects the progression of immune-related diseases. One of the hallmarks of immune cell activation and inflammation is the elevation of extracellular adenosine and increased expression of adenosine A2A receptors in activated immune cells. Adenosine strongly inhibits activation of helper T cells by elevating cAMP concentrations, which activate PKA and EPAC proteins to regulate cellular responses. Here we showed that adenosine strongly inhibits T cell accumulation rather than differentiating into functional subsets in the presence of polarizing conditions. Different aspects of adenosine mediated T cell suppression were phenocopied by specific activation of PKA and/or EPAC pathways. Adenosine can suppress phospho-activation of Akt pathway to promote T cell quiescence and to inhibit immediate downstream events after TCR stimulation. One of the targets for Akt is Foxo1. Foxo1 is known to suppress T cell proliferation. Our results showed that adenosine receptor stimulation reduced inhibitory phosphorylation of Foxo1 downstream of Akt. Foxo1 inhibitor, AS1842856, completely reversed the inhibition of T cell accumulation by adenosine signaling. Our results suggest that adenosine by activating PKA and/or EPAC pathways and by sustaining Foxo1 activation suppresses T cell activation and accumulation. These findings have important implications to develop novel interventions to regulate helper T cell responses in different pathological conditions.
Molecular mechanisms regulating CXCR4 mediated cytokine mRNA stability
Authors: Dinkel Brittney A , Kremer Kimberly , Hedin Karen E 
Published: January 2023

Abstract
T cell cytokine production has been shown to be dependent not only on T cell antigen receptor (TCR), but also the chemokine receptor CXCR4. Evidence in the literature suggests that, in addition to stimulating migration, CXCR4 might also more directly regulate T cell immune activation. Recent studies from our lab have revealed that TCR stimulation in the absence of SDF-1 induces association of TCR and CXCR4, and that this is inhibited by pretreating cells with the specific CXCR4 antagonist AMD3100. Cytokine production, particularly cytokines IL-2, IL-4, and IL-10, by TCR-stimulated cells was inhibited by using either AMD3100 or CXCR4 siRNA to prevent TCR-CXCR4 formation. Interestingly, TCR-CXCR4 was not required for activation of many TCR-stimulated signaling pathways or for cytokine gene transcription. Instead, TCR-CXCR4 formation during TCR-mediated T cell immune activation was required for the post-transcriptional stabilization of cytokine mRNAs through a Rac-1 mediated pathway. Our results suggest that targeting the TCR-CXCR4 complex formation or downstream signaling pathways may provide therapeutic targets to regulate cytokine production.
Leupaxin is serine phosphorylated and recruited to the immunological synapse upon TCR engagement in cytotoxic T lymphocytes (CTL)
Authors: Yao Shugang , Ostergaard Hanne 
Published: January 2023

Abstract

Background
Leupaxin is a member of paxillin family proteins that function as adaptor proteins in integrin signaling. Compared to other members, leupaxin is mainly expressed in leukocytes. However, the role of leupaxin in T cells is unknown.

Results
Leupaxin has been shown to be tyrosine phosphorylated in cells including tumor cells, fibroblasts and B cells. However we detected no leupaxin tyrosine phosphorylation in T cells. Instead, leupaxin showed mobility shift in response to TCR signaling. We demonstrated that the mobility shift was caused by serine phosphorylation and was ERK dependent. Although leupaxin is downstream of integrin receptors, LFA-1 stimulation did not induce leupaxin phosphorylation. Several potentially phosphorylated residues were mutated into alanine individually. When Ser54 was mutated, leupaxin no longer showed mobility shift, suggesting that leupaxin is phosphorylated at Ser54. Leupaxin was recruited to the immunological synapse and colocalized with LFA-1. Leupaxin has N-terminal leucine-aspartic acid (LD) domains and C-terminal LIM domains that contain four double zinc finger motifs. The LIM domains were dispensable for leupaxin localization at the contact zone. Leupaxin failed to be recruited to the immunological synapse after deleting LD2–4 domains, suggesting the importance of LD2–4 domains for the recruitment.

Conclusions
Leupaxin is not tyrosine phosphorylated but serine phosphorylated at Ser54 in response to TCR engagement. Leupaxin is recruited to the immunological synapse through LD2–4 domains. Studies are underway to determine the contribution of leupaxin to CTL adhesion, polarization and degranulation.
PKCθ Signaling in CD8 T cells regulates Notch1 and C-rel localization
Authors: Sherman Heather L , Ozay E. Ilker , Garrigan Daniel T. , Minter Lisa M. 
Published: January 2023

Abstract
The NF-kB family of transcription factors regulate many functions essential to T cells, including activation, proliferation, and survival. In resting T cells, certain NF-kB subunits are retained in inactive complexes in the cytosol. Following stimulation, phosphorylation events mediated by the IKK complex allows nuclear import of NF-kB and binding to target promoters. Using Jurkat CD4 T cells, it was shown that the NF-kB subunit, c-Rel, interacts with PKCq and anchors it to chromatin. Previous work in our lab demonstrated a physical and functional interaction between PKCq and the transmembrane receptor Notch1, in primary CD4 T cells, upstream of NF-kB activation. We further showed that inhibiting Notch1 signaling can attenuate symptoms in a model of acute graft-vs-host disease. Using this same model revealed that CD8 T cells require intact PKCq signaling to mediate disease, and data from the literature suggest CD8 T cells lacking PKCq are phenotypically similar to c-Rel-deficient CD8 T cells. Furthermore, we have discovered that PKCq, Notch1, and c-Rel localize differentially in CD4 and CD8 T cells following activation. Here we show that PKCq cellular localization regulates a novel interaction between Notch1 and c-Rel in CD8 T cells, which may constitute a crucial driver of graft-vs-host disease.
A Subset of Human Autoreactive CD1c-Restricted T Cells Preferentially Express TRBV4-1+ TCRs and Recognize Phospholipids
Authors: Guo Tingxi , Koo Edward M.Y. , Hirano Naoto 
Published: January 2023

Abstract
Unlike the highly polymorphic, peptide-presenting conventional MHC molecules, CD1 consists of a family of monomorphic, lipid-presenting proteins. Recent studies have revealed the molecular basis of mycobacterial lipid recognition by CD1a-c-restricted T cells. In addition to foreign lipids, subsets of CD1a-c-restricted T cells recognize self-lipids, which may have implications for human diseases such as autoimmunity and cancer. And yet, the molecular identity of these self-reactive T cells remains largely elusive. In this study, using a novel CD1c+ artificial antigen-presenting cell (aAPC)-based system, we have isolated human CD1c-restricted autoreactive T cells and characterized them at the molecular level. By employing the human cell line K562, deficient in MHC class I/II and CD1 expression, as a backbone, we generated an aAPC expressing CD1c as the sole antigen-presenting molecule with costimulatory molecules, CD80 and CD83. When stimulated with this CD1c+ aAPC endogenously presenting self-lipids, a subpopulation of primary human CD4+ T cells from multiple donors consistently upregulated CD154 (CD40L) in a CD1c-specific manner. These activated CD4+ T cells preferentially expressed TRBV4-1+ TCRs. Interestingly, TRAV usage and CDR3 sequences of these TRBV4-1+ T cells were diverse. Clonotypic analyses of the reconstituted TRBV4-1+ TCRs demonstrated that the heterogeneity of the CDR3 sequences greatly impacted the strength of CD1c-restricted autoreactivity. Furthermore, cell-free assays using recombinant CD1c loaded with distinct lipids identified several phospholipid species as potential self-ligands. These data provide new insights into the molecular identity of human autoreactive CD1c-restricted T cells.
Inhibition of Phosphatidylinositol-3-Kinase by Hibiscone C in T cells
Authors: Rawlings Jason S , Besley Caroline , Goess Brian 
Published: January 2023

Abstract
Inhibitors of Phosphatidylinositol-3-Kinase (PI3K) are of significant interest due to their ability to target an intracellular pathway, which regulates cell proliferation, growth, and migration. Mutations in PI3K can lead to an upregulation of the pathway, resulting in tumorigenesis, autoimmunity, and leukemia in the immune system, while a nonfunctional PI3K can result in immunodeficiency. Wortmannin, a known and potent PI3K inhibitor, possesses many unfavorable characteristics as a chemotherapeutic and immunotherapy drug such as its insolubility in neutral buffers, limited supply, and high toxicity in vivo, which have prevented its clinical application. The small molecule inhibitor Hibiscone C, synthesized from the readily available precursors acetone and isobutylaldehyde, shares the same structural chemistry at the binding site as Wortmannin, suggesting that it shares similar biological activity. Via analysis of phosphorylation of the downstream effector molecule Akt in activated T cells, we demonstrate that Hibiscone C also can irreversibly inhibit PI3K activity due to the presence of the furanosteroid ring, to which PI3K covalently binds in each molecule. As predicted by the lack of functional groups in Hibisconce C that are present in Wortmannin, Hibiscone C was found to be less potent of an inhibitor. Despite this reduced inhibitory potency, Hibiscone C serves as a minimalist structure with which to study the requirements for PI3K binding affinity and inhibition. This reduced structure can be manipulated in order to synthesize derivatives that can maximize potency, specificity, and solubility.
S1P1 regulation of pulmonary tertiary lymphoid tissues
Authors: Huppé Carole-Ann , Lecours Pascale Blais , Lechasseur Ariane , Morissette Mathieu C. , Marsolais David 
Published: January 2023

Abstract
Pulmonary tertiary lymphoid tissues (TLT) emerge as central regulators of antigen-specific responses in the lung and they have been associated with the severity of chronic respiratory diseases such as hypersensitivity pneumonitis (HP). The sphingosine-1-phosphate receptor S1P1 is a potent modulator of lymphoid organ biology and of the TH17 response, which is often associated with HP and TLT formation in the lung. We recently identified a novel immunogenic agent commonly found in the bioaerosols of working environments that induces an HP-like disease and potently stimulates TLT formation in the lung. We first aimed to determine if the ability of this agent to induce TLTs was related to their propensity to trigger a TH17 response in the lung. Secondly, we determined whether or not S1P1 acts as a modulator of pulmonary TLTs. Mice were exposed intra-nasally to Methanosphaera stadtmanae (MSS) for a 3-weeks time period in order to induce TLT formation. Upon MSS rechallenge, Il-17 expression was increased by 3 fold in the lung supporting the involvement of this cytokine in the current model. Among five homeostatic chemokines/proinflammatory cytokines tested, we found that CXCL13 was the most significantly increased by the MSS rechallenge (4.9-fold), supporting its involvement in TLT formation in response to MSS. S1P1 pharmacological agonism led to a 50% inhibition of Il-17 and Cxcl13 expression in the lung, which translated into a full inhibition of TLT expansion in response to the MSS rechallenge; and into a 77% alleviation of the pulmonary neutrophilic response. This study unravels a role for the S1P pathway in the regulation of TLTs as well as its interplay with the TH17 response in the lung.
Leptin Enhances TH2 and ILC2 Responses in Allergic Airway Disease
Authors: Zheng Handong , Zhang Xing , Castillo Eliseo F , Luo Yan , Liu Meilian , Yang Xuexian 
Published: January 2023

Abstract
Allergic asthma and obesity are the leading health problems in the world. Many studies have shown that obesity is a risk factor of development of asthma. However, the underlying mechanism has not been well established. In this study, we demonstrate that leptin, an adipokine elevated in obese individuals, promoted proliferation and survival of pro-allergic type 2 helper T cells and group 2 innate lymphoid cells and production of type 2 cytokines, which together contribute to allergic responses. Leptin activates mTORC1, MAPK and STAT3 pathways in TH2 cells. The effects of leptin on TH2 cell proliferation, survival and cytokine production are dependent on the mTORC1 and MAPK pathways as revealed by specific inhibitors. In vivo, leptin-deficiency led to attenuated experimental allergic airway inflammation. Our results thus support that obesity associated elevation of leptin contributes to the increased susceptibility of asthma via modulation of pro-allergic lymphocyte responses.
Accumulation of pulmonary vacuolated eosinophils in experimental allergic asthma that synergize with dendritic cells to induce Th-17 cells
Authors: Laumonnier Yves , Wiese Anna Valeska , Ender Fanny , Quell Katharina , Vollbrandt Tillman , König Peter , Köhl Jörg 
Published: January 2023

Abstract
Eosinophils are considered a homogenous cell population, contributing to the allergic asthma phenotype by promoting mucus hypersecretion and airway hyperresponsiveness. Here, we identified a previously unrecognized vacuolated CD11cdim eosinophilic population (vEOS) which accmulates exclusively in the lung tissue and in mediastinal lymph nodes but not in the airways in a house dust mite-mediated allergic asthma model and an IL-33 mediated pulmonary inflammation. In addition to the vEOS, we found classical SiglecF+CD11c− eosinophils (EOS) in the airways and the lung. Both cell types shared typical structural features of eosinophils and expressed CCR3. In contrast to EOS, vEOS expressed higher levels of the CD11a, CD11b and CD18 integrins. Immunofluorescence microscopy revealed that vEOS were located exclusively near arteries and airways, where they were in close contact with T cells. Moreover, vEOS took up antigen and expressed costimulatory molecules more efficiently than EOS. Consequently, in co-culture experiments with OVA-TCR transgenic T cells, vEOS and dendritic cells (DCs) drove T cell proliferation. However, in the presence of vEOS, T cells divided more frequently than in co-culture with DCs. Further, T cell activation in response to co-culture with vEOS together with DCs markedly enhanced IL-17 but not IL-13 production.
Collectively, our data demonstrate that vEOS are an important subset of eosinophils, which differ from EOS in their location and their ability to activate T cells. Further, they suggest that cross-talk between vEOS and DCs might be a crucial step for the induction of Th17 cells in allergic asthma. Thus, vEOS may serve as an important target for Th17-associated steroid-resistant asthma.
Impaired induction of Slc26a4 promotes respiratory acidosis and severe, steroid-resistant asthma
Authors: Kim Richard Yong Hoon , Pinkerton James W , Rae Brittany E , Mayall Jemma R , Brown Alexandra C , Ali Md Khadem , Goggins Bridie J , Essilfie Ama-Tawiah , Starkey Malcolm R , To Cuong , Bosco Anthony , Horvat Jay C , Hansbro Philip M 
Published: January 2023

Abstract
CO2 produced by systemic cellular respiration is hydrated into carbonic acid (H2CO3) that dissociates into H+ and HCO3−. These are transported in the plasma to the lungs where HCO3− is converted back into H2CO3 and CO2 that are expelled through breathing. Evidence suggests that dysfunction of the mechanisms that govern these processes may result in the development of respiratory acidosis (RA) and the cardinal features of severe, steroid-resistant (SSR) asthma. Reduced lung function, which occurs in SSR asthma, impairs removal of volatile H2CO3 and CO2, resulting in acid accumulation and increased arterial PaCO2. Patients with severe asthma often develop complications from increased PaCO2, which skews the PaCO2/HCO3− ratio resulting in increased H+ concentration and reduced pH. We developed three mouse models of respiratory infection and ovalbumin-induced SSR allergic airways disease (SSRAAD) that are highly representative of SSR asthma in humans. We used these models to show a role for impaired homeostatic acid-base balance in SSR asthma. All three infections suppress the induction of the expression of the chloride (Cl−)/HCO− pump, Slc26a4, in the airway mucosa in AAD. Importantly, SSRAAD is associated with increased levels of free H+ ions in bronchoalveolar lavage fluid. Administration of Slc26a4-specific siRNA in steroid-sensitive AAD, which mimics the effect of decreased Slc26a4 responses in SSRAAD, induced RA and steroid-resistant airway inflammation and AHR. Importantly, treatment of RA with NaHCO3 during infection-induced SSRAAD suppressed steroid-resistant AHR. Thus, we have identified a previously unrecognised role for deficient Slc26a4 responses that result in the development of RA and the pathogenesis of SSRAAD.