bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of new immune reagents for swine health, vaccine and disease studies
Authors: Manirarora Jean Nepomuscene , Bailey Michael , Renukaradhya Gourapura , Kenney Scott , LaBresh Joanna , Sang Yongming , Francis Ore , Wooldridge Linda , Lunney Joan 
Published: January 2023

Abstract
Current research efforts require a broad range of immune reagents; those available for pigs are limited. The goal of the US-UK Collaborative Swine Immune Toolkit Initiative is to generate priority immune reagents, based on international input, and pipeline them for marketing. UK researchers focus on mucosal targets, including production of monoclonal antibodies (mAbs) to chemokine receptors and IgE. Plans for mAbs to each 7-transmembrane G-protein coupled receptor (CCR3, CCR9, CCR10, CXCR1, CXCR6, F4/80) are to target peptides from extracellular loop 2 (ECL-2) based on threaded protein structures. These peptides have been designed for CCR3 and CCR9 and mAb plans are underway. US efforts are aimed at expression of soluble proteins and swine CD molecules, and production of panels of mAbs. The team has set up collaborations with commercial partners for protein expression and mAb production, and updated protocols to evaluate reagent specificity. New panels of mAbs reactive with IL-6, IFNβ and IFNγ are currently being evaluated. Immunizations are underway to produce panels of mAbs for CXCL10, CX3CL1 (fractalkine), IL-13, IL-17A, IL-17F, IFNω1 and IFNω5. Plans for mAbs to porcine CD1d using 4 synthetic peptides are in progress using CD1d knockout (KO) mice and screening on CD1d KO pig cells. Plans continue to use reagents produced to develop new assays for swine immune markers, e.g., multiplex assays and intracellular staining to provide the veterinary community with new commercial reagents and techniques for their research efforts. Tools and reagents generated by this project will undoubtedly advance swine immune, disease and biomedical research efforts. Supported by USDA ARS, NIFA grant #2015-67015-23216 and BBSRC grant BB/M028232/1.
WC1 is a hybrid gammadelta TCR coreceptor and pattern recognition receptor for pathogenic bacteria
Authors: Telfer Janice C , Le Page Lauren , Nandi Dipika , Buck Jessica , Boisvert Natalie , Gillespie Alexandra , Damani-Yokota Payal , Amir Maria , Yirsaw Alehegne , Baldwin Cynthia L 
Published: January 2023

Abstract
WC1 proteins are uniquely expressed on γδ T cells and belong to the scavenger receptor cysteine-rich (SRCR) superfamily. Present in variable, and sometimes high, numbers in most animals with adaptive immune systems, in cattle there are 13 distinct genes (WC1-1 to WC1-13). The structure of a multigenic WC1 gene array, with each WC1 protein containing multiple SRCR domains in its extracellular domain, has been conserved over millions of years of evolutionary time, suggesting that they act as a panel of pattern recognition receptors for conserved pathogen ligands. All bovine WC1 proteins can serve as coreceptors for the TCR in a tyrosine phosphorylation-dependent manner and some are required for the γδ T cell response to Leptospira. SRCR domain binding was directly correlated with γδ T cell response, as WC1-3 SRCR domains from Leptospira-responsive cells, but not WC1-4 SRCR domains from Leptospira-nonresponsive cells, bound to multiple serovars of two Leptospira species. Mutational analysis indicated that the active site for bacterial binding in one of the SRCR domains is comprised of amino acids in three discontinuous regions. In addition, recombinant WC1 SRCR domains with the ability to bind leptospires inhibited Leptospira growth. WC1 SRCR domains also bind to Borrelia burgdorferi and Mycobacterium spp. WC1 in other artiodactyls such as goat, sheep and swine are closely related, but show some divergence that may have been driven by natural selection by species-specific pathogens. Our data suggest that WC1 gene arrays play a multi-faceted role in the γδ T cell response to bacteria in multiple species, including acting as hybrid pattern recognition receptors and TCR coreceptors, and may function as anti-microbials.
Canine CD4+CD8+ double-positive T cells participate in immunity to Ehrlichia chaffeensis infection
Authors: McGill Jodi L , Boorgula Gunavanthi Devi Yadav , Wang Ying , Ganta Roman R 
Published: January 2023

Abstract
Ehrlichia chaffeensis is an obligate intracellular gram-negative bacteria and a frequent cause of severe and fatal tick-borne infection in people in North America. The reservoir host for E. chaffeensis is the white-tailed deer, while humans, dogs, coyotes and goats are regarded as common incidental hosts. In dogs, we have shown that E. chaffeensis establishes a chronic infection that persists for at least 42 days, while promoting development of Th1 and Th17 cellular responses and pathogen-specific humoral immunity. To date, little is known about the immune responses that correlate with control or successful clearance of the infection in dogs or humans. In dogs, a population of mature CD4+CD8+ double-positive (DP) T cells exist that have features of activated T cells and share some similarities to the CD4+CD8+ DP populations which have been described in pigs and humans. However, little is known about the function of these cells in dogs, particularly in the context of infectious diseases. Here, we demonstrate that canine CD4+CD8+ DP T cells expand significantly in the blood and spleen in response E. chaffeensis infection, and upregulate markers of T cell activation and antigen-experience. Recall responses after in vitro restimulation with E. chaffeensis host-cell free lysate showed that CD4+CD8+ DP T cells undergo multiple rounds of antigen-specific proliferation, produce IFNγ and IL-17, and upregulate expression of the cytotoxic molecules granzyme B and granulysin. Together, our results demonstrate that CD4+CD8+ DP T cells contribute to the immune response to E chaffeensis in the physiologic host, and suggest the importance of alternative lymphocyte populations in promoting protection and control of tick-borne diseases in the dog.
TAGLN2 inhibits Arp2/3 complex-nucleated actin branching and controls microvilli formation in T cells
Authors: Jun Chang-Duk , Kim Hye-Ran , Lee Kyung-Sik 
Published: January 2023

Abstract
Microvilli are abundant on the surfaces of circulating lymphocytes. However, factors that regulate microvilli formation on lymphocytes are not completely understood. Moreover, definitive roles of microvilli have not been fully addressed. Here we identified that TAGLN2—an actin-binding protein predominantly expressed in T cells— specifically localizes at the microvilli on T cells. Ectopic expression of TAGLN2 enhanced microvilli protrusion at the initial development of immunological synapse, and the opposite results were obtained by TAGLN2 depletion. Obviously, T-cell activation was influenced by the microvilli structures on T cells. Of interest was that TAGLN2 blocks Arp2/3 complex binding to actin filaments that leads to the inhibition of actin branch. Electron microscopy-based helical image analysis revealed that the main sites of TAGLN2 binding on actin share the Arp2/3 complex binding sites. These results demonstrate a novel function of TAGLN2 that contributes to enhance T-cell antigen recognition, and hence T-cell activation.
‘The role of phagocytic NADPH oxidase and innate immune cells in the immune activation in toluene diisocyanate (TDI)-induced lung inflammation’
Authors: CHENG CHIA YING , Shieh Chi-Chang K 
Published: January 2023

Abstract
Innate lymphoid cells (ILCs) have been known to play an important role in development of lymphoid tissues and in the initiation of inflammation at barrier surfaces in response to infection or tissue damage. Isocyanates are low-molecular-weight compounds noted for inducing an oxidant stress-dependent pulmonary inflammation, which is the leading cause of occupational asthma. Our previous studies indicated that phagocytic NADPH oxidase is an essential regulator in TDI-induced airway inflammation characterized by a server lung inflammation with both Th17 and Th2 immune components. In this study, we hypothesized that ILCs activation may play an important role in phagocytic NADPH oxidase-mediated downstream immune mechanism in TDI-induced airway inflammation. The wild type (WT) and phagocytic NADPH oxidase deficient (Ncf1−/−) mice were sensitized and challenged with 3% TDI to induce lung inflammation. Our results showed that type 2 inflammatory cytokines, including IL-33 and IL-5, increased in both WT and Ncf1−/−mice but more significantly in TDI-induced lung inflammation. Moreover, the pro-inflammatory cytokine IL-1b and IL-17A production was only induced by TDI-exposure in WT but not in Ncf1−/− mice. We also found that the population of lineage negative and CD45 positive cells was identified in inflammation lung tissue. Comparison of ILCs population between TDI-induced WT mice and Ncf1−/− mice showed that the number of ILC2s and ILC3s were significantly higher in WT mice. These results suggested that ROS may modulate the activation of ILC2s, ILC3s and may determine the Th2 and Th17 cytokines production in the TDI-induced lung inflammation.
IL-1R and IL-1RA differentially contribute to immunopathogenesis during fungal asthma
Authors: Godwin Matthew S , Blackburn Jonathan , Steele Chad 
Published: January 2023

Abstract
Severe asthma with fungal sensitization defines a subset of asthmatics with allergy to multiple fungal species, including Aspergillus fumigatus. We have previously shown genetic deficiency in or neutralization of IL-22 improves fungal asthma severity, which correlated with lower levels of IL-1α and IL-1β. Here, we show that bronchoalveolar lavage fluid (BALF) and induced sputum from human asthmatics sensitized to fungi demonstrate increased levels of IL-1α, IL-1β and IL-1 Receptor Antagonist (IL-1RA) compared to fungal negative asthmatics, suggesting that members of the IL-1 cytokine family may contribute to fungal asthma severity. To this end, we examined fungal asthma severity in mice deficient in IL-1R and IL-1RA with the hypothesis being that severity would be decreased and increased in these animals respectively. Surprisingly, the absence of IL-1R signaling had no impact on biomarkers of asthma severity (i.e. levels of proallergic CCL17, CCL22, and IL-33), yet Il1r−/−mice demonstrated better lung function compared to WT mice. Interestingly, the absence of IL-1R signaling resulted in a profound defect in the production of IL-22, suggesting a mechanism of improved lung function in Il1r−/− mice. In direct contrast, Il1rn−/− mice exhibited elevated proallergic biomarker levels, but no changes in the levels of IL-22. These preliminary results identify differential roles for IL-1R signaling (IL-22 mediated immunopathogenesis) and IL-1RA signaling (type 2 mediated immunopathogenesis) in fungal asthma severity.
Inhibition of tight junction protein ZO-1 expression by estrogen in human gut mucosa
Authors: Zhou Zejun , Gilkeson Gary , Jiang Wei 
Published: January 2023

Abstract

Background
Females have stronger cellular and humoral immunities compared to males. Sex hormones are playing roles in these differences; however, the role of sex hormones in human gut permeability is essentially unknown.

Methods
The expression of tight junction ZO-1, estrogen receptor (ER)-α and β in male and female primary gut tissues were analyzed using immunohistochemistry. The inhibition of estrogen on ZO-1 was evaluated in primary gut tissues by qPCR. ZO-1 expression and NF-κB activation in Caco-2 cells was evaluated by western blot, immunofluorescent staining and reporter activity assays. Data are presented as median (interquartile range [IQR]).

Results
The expression of ZO-1 and ER-β but not ER-α were present in both male and female gut tissues, the median immunofluorescence intensities (MFI) of ZO-1 expression were 2.48 (2.40–3.11) and 3.59 (3.11–4.23) for females and males respectively (P = 0.02, non-parametric Mann-whitney test). No sex difference in MFI expression of ER-β was found (P = 0.79). After 24 h incubating with physiologic dose of estrogen (200 pg/mL) in primary gut tissues in vitro, the median ZO-1 mRNA relative expression to GADPH by qPCR was 1.002 (1.000–1.003) and 0.80 (0.21–0.84) for treatment with medium alone and estrogen respectively. The median MFI expression of ZO-1 in Caco-2 cells assessed by microscopy was 0.97 (0.77–1.14) and 1.16 (0.76–1.37) for treatment with estrogen and medium alone respectively (P = 0.01). The median luciferase activity of NK-κB in Caco-2 cells assessed by NK-κB reporter activity assay was 0.58 (0.53–0.60) and 0.98 (0.87–1.05) for treatment with estrogen and medium alone respectively (P = 0.002).

Conclusion
Estrogen inhibits the ZO-1 expression and NF-κB activity in human gut mucosa.
A novel method of assessing ASC specks and caspase-1 activity by imaging flow cytometry
Authors: Nagar Abhinit , DeMarco Richard A , Harton Jonathan A 
Published: January 2023

Abstract
NLRP proteins interact with the adaptor ASC to recruit and activate caspase-1-dependent maturation of IL-1β and IL-18 in a multiprotein ‘inflammasome’ complex. Inflammasome activation occurs in response to structurally diverse agonists triggering NLRP activation leading to rapid localization of NLRP:ASC complexes in a typically singular perinuclear ‘speck’. At present, formation of inflammasome specks can be assayed by microscopy, which is time consuming, or a flow cytometric analysis technique evaluating altered width and height of ASC pulse (TOFIE), which provides no visualization of the speck. We have developed a technique utilizing the advantages of imaging flow cytometry that combines high-throughput sample collection and high magnification (60×) microscopic visualization. This technique not only detects the presence of ASC specks but also determines the distribution of active caspase-1 within the cell using the fluorescent caspase-1 substrate FLICA. This assay allows for high-throughput imaging of ASC-containing specks and characterization of these specks in terms of number and area. Additionally, this technique also determines the location of active caspase-1 and enumerates aggregates of active caspase-1. This method is more sensitive than the present flow cytometric method, TOFIE, which fails to differentiate between active inflammasome specks and false positive signals due to accumulation/aggregation of ASC. The capacity of this method to detect and quantify native as well as reconstituted inflammasome specks with associated caspase-1 activity renders it more powerful for quantifying the impact of mutations on the molecular interaction of NLRP proteins with ASC and accompanying caspase-1 activation.
Aryl hydrocarbon receptor: a novel target for the anti-inflammatory activity of statin therapy
Authors: Turco Antonella , Scalisi Giulia , Gargaro Marco , Pirro Matteo , Fallarino Francesca 
Published: January 2023

Abstract
Statin therapy is associated with an attenuation of the systemic inflammatory response. Aryl hydrocarbon receptor (AhR) is believed to play an active role in the control of inflammation. Specific statins promote the transcription of Cyp1a1, which induction involves AhR activation1. We investigated whether AhR might be involved in the anti-inflammatory effect of statins treatment.
We performed luciferase assay to test the ability of murine hepatocytes overexpressing AhR and the xenobiotic responsive elements (XRE) and the ability of murine embryonic fibroblast AhR KO, transfected with different mutants of AhR to activate receptor in response to statins. Lipopolysaccharide (LPS)-treated RAW 264.7 macrophage cells were tested for the expression of AhR and for pro-inflammatory/anti-inflammatory response to different statins. Activated AhR WT and KO macrophages were tested for the expression of M1 and M2 markers and for pro-inflammatory/anti-inflammatory cytokines production. Luciferase assays showed that atorvastatin and pravastatin activated AhR, but no AhR mutants. LPS treatment of RAW 264.7 cell line induced AhR. Exposure of RAW 264.7 to both pravastatin and atorvastatin concomitantly to LPS reduced IL-6 production and increased IL-10 secretion. Atorvastatin and pravastatin potentiated arginase expression and anti-inflammatory response in activated macrophages in AhR dependent manner.
These data show that atorvastatin and pravastatin activate AhR, attenuate the inflammatory response induced by LPS and promote M2 phenotype in AhR dependent manner. AhR might represent a mediator of the anti-inflammatory effects of statin treatment.
Attenuation of Staphylococcal enterotoxin-B (SEB)-induced lung injury (ALI) by resveratrol results from miR-193a down-regulation and alterations in metabolic profile
Authors: Alghetaa Hasan F , Mohammed Amira K , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Resveratrol, (RES), a phytoalexin, is well-known for its anti-inflammatory and anti-oxidant properties. SEB, a superantigen, is known to trigger ALI and cause mortality. In the current study, we tested the effect of RES in a dual-dose model of SEB exposure that triggers ALI and causes 100% mortality in C3H/HeJ-mice. The data revealed RES attenuated SEB-induced ALI and prevented mortality. Forty eight hours post-SEB exposure, lung-infiltrating mononuclear cells were tested for microRNA expression profile to determine the epigenetic regulation by resveratrol. SEB-activated splenocytes were pre-treated with 50 μM of RES or vehicle for metabolic profile analysis by measuring oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). We also noted significant decline in miR-193a in the lungs of RES-treated SEB group, which targeted and caused an increase in TGFβ2 and TGFβR3, potent inhibitors of T-cell proliferation, by using RT-PCR and validation by transfection studies. RES-induced downregulation of miR-193a also influenced the activity of mechanistic target of rapamycin (mTOR) as well as pyruvate kinase muscle isozyme2 (PKM2) genes, and caused RES-treated SEB-activated T cells to be quiescent metabolically in comparison to the energetic vehicle-treated SEB-activated T cells. Together, RES caused inhibition in the proliferation of SEB-activated T-cells by alterations in miR expression and metabolic profiles. (Supported by NIH grants P01AT003961, R01AT006888, R01ES019313, R01MH094755, P20GM103641 and R01AI129788).
The role of microglia and monocyte-derived macrophages in Toxoplasma gondii infection of the brain
Authors: Thompson Jeremy , Hayes Nikolas , Gevaria Anjana , Harris Tajie 
Published: January 2023

Abstract
In the brain, microglia are the resident immune cells. Despite their location in the brain parenchyma and expression of machinery capable of responding to pathogens, little is known about the role microglia play during infection. Toxoplasma gondii is an intracellular, protozoan parasite that establishes a chronic infection in the brain. T cell-derived IFN-g is central to control parasite replication and prevent host mortality by activating anti-parasitic mechanisms. In addition, during brain infection monocyte-derived macrophages (MDMs) enter the brain. Deciphering the contribution of these two cell populations during T. gondii infection has been confounded by their phenotypic and morphologic similarity. However, these cells likely contribute differently to parasite control. Using a reporter mouse to label microglia we have found that MDMs are the primary producers of nitric oxide, a key anti-parasitic molecule. Therefore, to explore the role of microglia and MDMs in the control of T. gondii in vivo we are taking advantage of constitutive or inducible cre recombinase activity driven from the CX3CR1 promoter which allow both microglia and MDMs or solely microglia, respectively, to be targeted genetically. By removing the gene STAT1 from these cells we have rendered them unable to upregulate key IFN-g induced genes necessary for controlling T. gondii. Mice in which MDMs and microglia are unresponsive to IFN-g are unable to control parasite replication and succumb to infection by 19dpi. Conversely, mice in which only microglia are deficient in IFN-g signaling do not succumb to infection and resemble infected control mice. These results suggest that microglia are not a key cell type responsible for controlling T. gondii in the CNS.
The immune environment dictates the dominance of licensed versus unlicensed natural killer cell subsets during viral infection
Authors: Aguilar Ethan George , Zamora Anthony Elston , Dunai Cordelia , Khuat Lam , Murphy William J 
Published: January 2023

Abstract
Natural Killer (NK) cells are innate lymphocytes involved in anti-viral and anti-cancer responses. They can be functionally and phenotypically divided based on expression of inhibitory receptors capable of binding to self-MHC class I, a concept termed licensing/education. NK cells capable of binding to self-MHC (licensed) have been shown to be more functionally competent compared to those that cannot (unlicensed). It is still unclear which subset is more critical during an anti-viral response. Using two models of murine cytomegalovirus infection (MCMV), we show distinct subsets of NK cells that can both directly and indirectly contribute to the anti-viral response. We have previously shown that after hematopoietic stem cell transplantation (HSCT), the licensed effector NK population was predominantly responsible for resistance to MCMV, which was in part due to the absence of T cells present immediately after transplant. However, MCMV infection in non-HSCT mice yielded an opposite pattern with the unlicensed NK subset augmenting resistance to MCMV. We found preferential localization patterns of the different NK subsets which correlated with increased antigen specific T cell expansion and viral clearance. Interestingly, by depleting the different NK subsets in HSCT vs non-HSCT mice, we demonstrated differences in subset roles. The unlicensed NK cells, which promote antigen specific T cells, are more critical during normal conditions, while the licensed NK cells serve a better direct anti-viral role following HSCT where T cells take longer to repopulate. Thus, the contributions of these different NK subsets and their dependence on the immune environment should be taken into consideration when applying NK cells therapeutically.