bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeted Gene Editing Restores Regulated CD40L Expression and Function in X-HIGM T cells.
Authors: Hubbard Nicholas Wesley , Hagin David , Sommer Karen , Song Yumei , Khan Iram , Clough Courtnee , Ochs Hans D , Rawlings David J , Scharenberg Andrew , Torgerson Troy R 
Published: January 2023

Abstract
Loss of CD40L expression or function results in X-Linked Hyper-IgM Syndrome (X-HIGM), characterized by recurrent infections due to impaired immunoglobulin class-switching and somatic hypermutation. Previous attempts using retroviral gene transfer to correct murine CD40L expression restored immune function; however, treated mice developed lymphoproliferative disease, likely due to viral-promoter dependent constitutive CD40L expression. These observations highlight the importance of preserving endogenous gene regulation in order to safely correct this disorder. Here we report efficient, on-target, homology directed repair (HDR) editing of the CD40LG locus in primary human T cells using a combination of a TALEN-induced double-strand break and a donor template delivered by recombinant Adeno-Associated Virus (rAAV). HDR mediated insertion of a coding sequence (GFP or CD40L) upstream of the translation start site within Exon 1 allowed transgene expression to be regulated by endogenous CD40LG promoter/enhancer elements. Additionally, inclusion of the CD40LG 3′-untranslated region (3′-UTR) in the transgene preserved post-transcriptional regulation. Expression kinetics of the transgene paralleled that of endogenous CD40L in unedited T cells, both at rest and in response to T cell stimulation. The use of this method to edit X-HIGM patient T cells restored normal expression of CD40L and CD40-muIg binding, and rescued IgG class switching of naïve B-cells in vitro. These results demonstrate the feasibility of engineered nuclease-directed gene repair to restore endogenously regulated CD40L, and the potential for its use in T cell therapy for X-HIGM syndrome.
Mesothelin Vaccination for the Prevention of Ovarian Cancer
Authors: Scholler Nathalie , Stein Paul , Repellin Claire , Kamat Kalika , Harrison M. Travis , Shoemaker Robert , Sambucetti Lidia 
Published: January 2023

Abstract
Survival of ovarian cancer patients with high-grade serous carcinoma (HGSC) remains low. We hypothesized that a tumor antigen-targeted vaccine capable of eliciting a robust cell-mediated immune response may substantially improve women’s health. Mesothelin is a glycosylphosphatidylinisotol (GPI)-anchored, conserved protein that is overexpressed by ovarian, lung and pancreatic cancers. We designed mesothelin vaccines containing cyclic dinucleotide (CDN)-based adjuvants or alum-based adjuvants to trigger CD8 T cell activation. Anti-mesothelin antibody titers were stable or slightly increased after 2 vaccine boosts, and the highest and most stable titers were obtained with CDN-based adjuvants (meso/CDN). In addition, splenocytes from all mice receiving CDN-containing vaccines secreted mesothelin-specific IFNγ. Next, immunized mice were injected orthotopically with luciferase-transduced mouse ovarian cancer cells (ID8). In vivo imaging performed 3 weeks after tumor injections revealed that most of the mice from the groups immunized with Alum-based vaccines (92%) presented a positive signal of bioluminescence, which corresponds to the routine take rate of the ID8 tumors. In contrast, only 1/3 of the mice in the meso/CDN group showed a positive signal compared with 2/3 of the mice immunized with the control CDN-only vaccine. These results suggest that the meso/CDN vaccine regimen delayed tumor establishment. Longer observation of the mice is ongoing to determine whether the vaccination regimen can prevent tumor development. We tentatively conclude that ovarian cancer prevention may be possible through vaccination against mesothelin tumor antigen in combination with adjuvants that trigger cellular immunity.
In vitro treatment of neonatal foal alveolar macrophages with CpG 2142 enhances their killing capacity of R. equi
Authors: Bordin Angela Ilha , Gressler Leticia Trevisan , Rocha Joana , Brake Courtney , Tanner Amelia , Juillard Veronique , Cohen Noah 
Published: January 2023

Abstract
Pneumonia caused by Rhodococcus equi, a facultative, intracellular bacterium that replicates within macrophages, is an important cause of disease and death in immunocompromised hosts. No vaccine is yet available, and control is based on treatment with antibiotics. The emergence of antibiotic resistance and the lack of effective alternative antimicrobials indicate novel approaches to control are needed, such as host-directed strategies. Cytosine-phosphate-guanine oligodeoxynucleotides (CpGs), Toll-like receptor (TLR) 9 agonists, have immunomodulatory function both in vitro and in vivo in foals. Our objective was to evaluate the impact of in vitro treatment of neonatal foal alveolar macrophages (AMs) with CpG 2142 during the first month after birth. Using broncho-alveolar lavage, AMs were collected from 6 foals at ages 2, 14, and 26 days. Effects of age and treatment on the intracellular killing of R. equi by foal AMs were determined using the ratio of colony-forming units at 48 hours (T48) to post-phagocytosis (T0) (i.e, T48:T0). Foal AMs had significantly (P &lt; 0.05) greater killing capacity at 14 and 26 days than at 2 days of age. Relative to media control, CpG 2142 increased the killing capacity of R. equi by foal AMs at ages 2, 14, and 26 days by approximately 2, 10, and 8 fold (P &lt; 0.05), respectively. Notably, CpG 2142 clearly enhanced killing of R. equi by foal AMs even at age 2 days, when neonatal foals are known to lack a fully functional immune system. CpG 2142 can stimulate innate immune cells including AMs to kill intracellular pathogens more efficiently, and might be effective when nebulized for host-directed therapy against pneumonia caused by R. equi and similar intracellular pathogens such as Mycobacterium tuberculosis.
Maternal vaccination with dPNAG protects foals against the intracellular pathogen Rhodococcus equi
Authors: Rocha Joana Nunes , Cywes-Bentley Colette , Bordin Angela I. , Brake Courtney N. , Pier Gerald B. , Cohen Noah D. 
Published: January 2023

Abstract
Rhodococcus equi is a facultative intracellular pathogenic bacterium that causes a form of pneumonia similar to tuberculosis in foals and immunocompromised humans. A vaccine against R. equi is lacking. We studied the efficacy of maternal vaccination with deacetylated poly-N-acetyl glucosamine (dPNAG) oligosaccharide conjugated to tetanus toxoid to protect foals against R. equi. Pregnant mares (n=9) were randomly assigned to intramuscular injection 6 and 3 weeks prior to foaling with either vaccine (n=5) or saline (n=4). Foals were infected intrabronchially at age 28 days with 1 × 106 of a single strain of live, virulent R. equi. Foals were examined twice daily for clinical signs of R. equi pneumonia (viz., coughing, tachypnea, dyspnea, temperature &gt;103°F), and weekly by thoracic ultrasonography. Blood samples were collected at age 2, 14, 28, 42, 56, and 84 days from all foals to detect antibodies by ELISA against dPNAG and native PNAG in serum and IFNγ production from peripheral blood mononuclear cells (PBMCs). The proportion of foals with pneumonia was significantly (P&lt;0.05) greater among the unvaccinated mares (3/4) than vaccinated mares (0/5). Titers of dPNAG and PNAG antibodies were significantly (P&lt;0.05) higher at all ages for foals of vaccinated mares. IFNγ produced by PBMCs stimulated with R. equi antigen was significantly greater (P&lt;0.05) at age 28 days (prior to infection) in foals of vaccinated mares. Results indicate the dPNAG vaccine protected foals against the intracellular pathogen R. equi and efficacy against R. equi was likely mediated by maternally-transferred antibodies and enhanced CMI responses, and contribute to a growing body of evidence for a key role of antibodies in protecting against intracellular pathogens.
Identification of helitron sequences in the coral genome and implications for host defense.
Authors: Garrison Keith E 
Published: January 2023

Abstract
Helitrons are mobile elements found in the genomes of numerous plant and animal species. Helitrons can mobilize flanking sequences when they transpose, and so have the potential to duplicate genes and remodel genomes. The stony coral Acropora has an expanded immune gene repertoire in comparison to the closely-related anemone species Nematostella, especially in the number of TLR and IL-1R genes. Mechanisms behind the expansion are not currently known. Although helitron sequences have been identified in Nematostella, no helitrons have been identified in Acropora. BLAST searches against the Acropora genome using Nematostella amino acid sequences as queries revealed seven candidate Acropora helitrons. Six of the seven Acropora candidates had large open reading frames in the forward direction. One candidate element was split between two adjacent reading frames. Conserved domain searches were performed within the open reading frames using the databases at NCBI. As expected, Helitron N-like and replicative protein A (RPA) domains were shared between Acropora and Nematostella elements. UPGMA trees were constructed using highly conserved regions, and one candidate Acropora element clustered more closely with the Nematostella reference helitron than with other Acropora sequences. Interestingly, P-loop domains were common to cnidarian helitrons and a subset of plant helitron reference sequences containing plant disease resistance gene motifs in regions overlapping with a nucleotide-binding domain critical for resistance gene function. Together, these studies will help to determine if helitrons played a role in the expansion of the Acropora immune gene repertoire.
Lutein Ameliorates Streptozotocin-Induced Cardiac Oxidative Stress in Rats
Authors: ALREJAIE SALIM , ALHUSAINI KHALID 
Published: January 2023

Abstract

Background and Aims
A number of clinical studies and animal models have established that hyperglycemia is the leading cause for the progression of diabetic cardiovascular complications. An elevated glucose levels induces structural and functional changes in the cardiomyocytes through the activation of several signal transduction pathways, one of them is lipid peroxidation. The present study was undertaken to evaluate the protective effects of lutein against cardiac oxidative stress in streptozotocin (STZ; 65 mg/kg)-induced diabetic rats.

Method
Lutein content (40, 80 and 160 mg/kg) diets were supplemented to diabetic rats for six weeks. Thiobarbituric acid reactive substances (TBARS) and glutathione (GSH) levels, and superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST) activities were measured in cardiac tissue. Histopathological changes were observed in cardiac tissue.

Results
Lutein dietary supplementations significantly inhibited the diabetic induced increased in heart levels of TBARS and enhanced the decreased levels of GSH in dose dependent manner. Markedly inhibited the cardiac enzymatic activities of SOD, CAT, GPx, GR and GST in diabetic rats were significantly increased in lutein diet supplemented diabetic animals compared to diabetic vehicle group.

Conclusion
These findings suggest that lutein has the potential to ameliorate STZ-induced toxicity in cardiac tissue probably through its antioxidant properties.
Comparative efficacy of equine bone marrow derived stem cell culture protocols.
Authors: Pullen Nicholas A. , Ryan Joan M. , Schiltz Paul M. 
Published: January 2023

Abstract
Across species the differentiation and propagation of various stem cell subsets are essential processes for research into immune cell and regenerative biology, and the development of new clinical opportunities. Especially in the equine realm there is an ever-growing interest in the effective and replicable generation of multipotential mesenchymal stem cells (MSC) for clinical resolution of intra-articular pathologies. Bone marrow is a desirable equine MSC source due to ease of access and the inclusion/richness of hematopoietic stem cells (HSC) that can also be studied from the same patient; however, there are numerous competing protocols for the differentiation of MSC, mostly concerning the initial isolation and enrichment steps from patient samples. Using sternebral BM biopsies we compared the efficacy of MSC purification and culture protocols including: carbohydrate/density gradient cell centrifugation, erythrocyte lysis, and cell settling/immediate subculture methods. Furthermore, with substantially fewer standardized equine-specific reagents available for the differentiation of HSC, we performed preliminary culture assessments of cross-reactivity for cytokines from non-equine sources, specifically IL-3 and SCF. The data presented here contribute to clinical and research decision-making with respect to MSC under differing circumstances of sample size, availability, scale of return, and time in addition to shedding light on research opportunities for growth and differentiation of equine HSC lineages.
Reduced memory responses to vaccination in cows naturally infected with bovine leukemia virus.
Authors: Frie Meredith , Sordillo Lorraine , Bartlett Paul , Coussens Paul 
Published: January 2023

Abstract
Bovine leukemia virus (BLV) is an extremely prevalent retrovirus infecting US dairy herds. Approximately 83% of US dairy herds are BLV-positive, and the within-herd infection rate is typically between 25–50%, with the prevalence increasing as the herd size increases. Historically, BLV infection has been considered benign, leading to a lack of control for its spread. However, recent research suggests that BLV infection negatively impacts the health of the host immune system. If BLV impairs host immunity, it would have a large economic impact on the dairy industry. To assess the immune competence of dairy cattle naturally infected with BLV, both BLV- and BLV+ cows were boosted on their regular schedule with BoviShield Gold FP5 L5 HB. Samples were collected over 28 days to characterize the strength of the immune response to the vaccine stimulus. BLV+ cows produced significantly lower titers of antigen-specific antibodies in comparison to healthy, BLV- cows. In vitro assays suggest that the difference in antibody titers is related to lower interferon-γ production from T cells in response to antigenic stimulation. Our results support the hypothesis that BLV+ cows have a reduced immune response to a commonly used commercial vaccine, which suggests that BLV+ cows may be less protected by vaccines and, thus, at an increased risk of developing secondary infections.
Variegated expression of members of the WC1 pathogen recognition receptor and co-receptor multi-gene family on γδ T cells
Authors: Damani-Yokota Payal , Telfer Jancie C , Baldwin Cynthia L 
Published: January 2023

Abstract
Cells of the immune system recognize disease-causing pathogens and respond in a manner to stop the infection. While we know how conventional cells of the immune system do this, for some non-conventional cells such as γδ T cells this process is less clear. We have shown that bovine γδ T cells bear lineage-specific transmembrane glycoproteins, known as WC1, are coded by a multigenic family and that these molecules function both as pattern recognition receptors (PRR) and signaling co-receptors for cellular activation. For example, while a subpopulation known as WC1.1+ γδ T cells respond early and participate in a recall response to the spirochetes Leptospira interrogans and Leptospira borgpetersenii, other subpopulations that express a different set of the WC1 genes do not respond. Other experiments showed that WC1 molecules on γδ T cells that respond to Leptospira bind the bacteria and are involved in cell activation. Thus, we hypothesize that WC1 is essential for the recognition of bacterial pathogens by γδ T cells and that the WC1 family members expressed by a particular cell will determine its ability to respond to an infection. We are now evaluating the distribution of WC1 gene expression among individual T cells to understand how subpopulations respond to specific pathogens. WC1 expressing γδ T cell clones were generated by infection with Theileria parva or by generation of long-lived memory cells to Leptospira Ag. To date, our findings suggest that the number of WC1 family members per an individual γδ T cell is restricted and stably expressed.
Stress perception differentially regulates the inflammatory response in the CNS to a neurotropic viral pathogen
Authors: Steel Christina D , Machida Mayumi , Lundberg Patric S , Wellman Laurie , Sanford Larry D , Ciavarra Richard P 
Published: January 2023

Abstract
Intranasal application of neurotropic vesicular stomatitis virus (VSV) replicates in olfactory receptor neurons and then rapidly invades the olfactory bulb (OB). VSV has an established time course of progression through discrete brain regions and of onset and recovery from encephalitis. Coincident with viral progression is a bimodal brain cytokine response composed of multiple chemokines and cytokines responsible for the prominent neuroinflammatory response. Stress can have significant, but poorly understood, effects on immune function and even less is known on how stress perception may impact immune competency. Therefore, VSV infected mice were exposed to either controllable (modeled by escapable foot shock, ES) or uncontrollable (modeled by inescapable foot shock, IS) stress prior to infection and for 1–3 days post infection (PI). OB cytokine responses were initially blunted and then amplified and dysregulated at days 1 and 3 PI, respectively. With time, dysregulated responses in ES mice were associated with increased pathogenecity. Ingenuity Pathways Analysis (IPA) identified miRNA146a as an upstream regulator of this response and qPCR found elevated OB levels (~5 fold) in ES versus IS cohorts at day 1 PI. Surprisingly, brains from mice that had long ago cleared VSV still contained an infiltrate containing CD103+ cells, a marker associated with long-term brain T resident memory cells. To our knowledge, this is the first time that stress perception has been shown to differentially modulate neuroinflammation. Our data also suggest the microRNA likely responsible for the observed stress-induced changes in the antiviral immune response to a neurotropic viral pathogen.
Development of interferon mimetic peptides having antiviral activity
Authors: Mujtaba Mustafa G 
Published: January 2023

Abstract
Current treatment options are limited for ameliorating viral infections. Interferons are involved in immune responses to viral infections; however, these interferons are not induced by some viral infections and, furthermore, can have toxic side effect when given at high doses as a therapeutic. The objective of this research is to design peptide mimetics of interferons that are not toxic to cells but have antiviral activities similar to interferons. Eleven peptides were designed based on interferon sequences and their receptor binding sites. Their activity on cell proliferation, toxicity, and antiviral effects against vesicular stomatitis virus (VSV) challenge of HeLa cells were measured. Results show that none of the peptides had significant antiviral activity when peptides were preincubated or incubated at the same time with the cells before VSV challenge. However, one peptide (Bact2) did have antiviral activity at 60ug/mL when this peptide was preincubated with the VSV virus for 24 hours before the HeLa cells were added to the mixture, suggesting that the Bact2 peptide directly bound to the VSV virus for its antiviral effect. Further analysis of the toxicity of the Bact2 peptide incubated alone with HeLa cells showed no cellular toxicity at concentrations of 60ug/mL or lower. Thus, the results from this study show that short peptides could be developed to directly neutralize viruses and interfere with their infectivity of cells.
RIG-I-like Receptor-mediated Signaling Regulated by NSs through Its Interaction with MAVS in a Novel Bunyavirus Infection
Authors: Xing Zheng , Zhu Lili , Qi Xian , Wu Xiaodong , Cardona Carol J 
Published: January 2023

Abstract
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease, which is caused by SFTS virus (SFTSV), a novel bunyavirus transmitted by ticks from animals. The disease is characterized by high fever and drastic reduction of leukocytes and platelets with high mortality rates in severe cases. We have reported that SFTSV suppresses host antiviral immunity and the viral nonstructural protein NSs plays a role in the immune evasion by interacting with downstream interferon signaling effectors, which is probably significant in virus pathogenesis. We furthered our study and demonstrated in this report that NSs interacted with mitochondrial antiviral signaling protein MAVS, the essential linker for host innate responses. NSs formed unique inclusion bodies (IBs) in the cytoplasm of infected cells and, through NSs and MAVS interaction, MAVS was translocated into the IBs in both transfected and infected cells. The interaction was specific and NSs did not interact with RIG-I or MDA5. Co-localization of NSs and MAVS trapped a significant portion of MAVS in the cytoplasmic IBs and weakened the functions of MAVS in activating downstream NFκB and IRF3 signaling. We examined host responses in NSs-overexpressing or infected cells and observed significant reduction of pro-inflammatory cytokine (IL-6, IL-8, Fas) and IFNs (IFN-α, IFN-β). Our data suggest that MAVS is essential to anti-SFTSV response since the knockdown of MAVS promoted viral replication via decreased IFN-β production, which could be reversed by overexpression of MAVS in the cells. This report demonstrates another player key to host responses, targeted by NSs of SFTSV through sequestration into viral IBs, resulting in inhibited IFN and NF-κB responses.
A central role for Type I IFN in the induction of Th2 responses by dendritic cells
Authors: Webb Lauren M , Lundie Rachel J , Borger Jessica G , Cartwright Adam N , Cook Peter C , Brown Sheila L , Jackson-Jones Lucy , Phythian-Adams Alex T , Davis Daniel M , MacDonald Andrew S 
Published: January 2023

Abstract
Although dendritic cells (DCs) are critical for induction of Th2 immunity against helminths or allergens, relatively little is known about how they become activated and function in response to Th2-polarizing antigens. We have discovered a previously unrecognized role for Type I IFN (IFN-I) in the optimal activation and function of DCs following exposure to strongly Th2-polarizing antigens. To date, IFN-I has primarily been associated with anti-viral immunity, and its role in Th2 settings is currently unclear. DCs exposed to total egg antigens from the parasitic helminth Schistosoma mansoni, or the primary immunostimulatory component of S. mansoni eggs, omega-1, produced IFN-I in vitro. IFN-I was also detected in response to the common allergen house dust mite (HDM). DCs lacking the IFN-I receptor displayed a dramatically impaired ability to induce Th2 cytokines in vivo, but unimpaired ability to support CD4 T cell polarization in vitro. Further, Th2-promoting DCs depended on IFN-I signaling for optimal activation, efficient migration to the draining LN, and effective localization within the T cell zone. In vivo challenge of naïve mice with S. mansoni eggs or HDM in the absence of the IFN-I receptor induced significantly reduced levels of Th2 cytokines compared to controls. Together, our data suggest a key role for IFN-I to enable Th2 induction by DCs against Th2 Ag in vivo. Future work will address the wider role of IFN-I in Th2 inflammation, including during patent helminth infection in vivo.