bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterizing the T-cell survival and differentiation during priming with downregulated γ-c cytokine/Jak3 signals (50.34)
Authors: Khattar Mithun , Wang Guohua , Miyahara Yoshihiro , Guo Zhiyong , Chen Wenhao , Stepkowski Stanislaw 
Published: January 2023

Abstract
T-cells play a central role in cell mediated immune responses to foreign antigens. Optimal T-cell activation and expansion requires antigen recognition via cognate T-cell receptors (TCRs) along with co-stimulatory and cytokine signals. The cytokine signals that lead to T-cell activation and proliferation, involve binding of common gamma-chain cytokines to their cognate receptors, that in turn activates Janus tyrosine kinase (Jak) 3, inducing cytokine gene transcription via the Jak/Stat (Signal Transducers and Activators of Transcription) pathway. However, it is currently unknown as to whether these paradigms of signaling during T-cell priming are universal to all T-cell subsets and lineages or unique to each. In the current study, we have examined the role of γ-c cytokine/Jak3 axis of signaling in survival and differentiation of different T-cell subsets during early phases of priming. We show highly reduced survival of CD8+ T-cells under cytokine/Jak3 inhibiting conditions during activation, whereas CD4+ T-cells were fairly more resistant. Loss of Jak3 signaling also inhibited the Treg differentiation while promoting pro-inflammatory Th17 cells thus, suggesting discrete and specific roles of this signaling pathway in different T-cell subsets and lineages. These findings contribute to our understanding of T-cell activation in different T-cell lineages as well as their differentiation into subsets, and are important for designing therapeutic approaches of the immune responses.
MEK Kinase 1 signaling regulates Natural Killer T cells (50.38)
Authors: Gallagher Ewen 
Published: January 2023

Abstract
MEK Kinase 1 (MEKK1) is of critical importance in controlling the thymus-dependent immune response by regulating Mitogen-Activated Protein Kinase (MAPK) activation from many receptors. Natural Killer T (NKT) cells are a distinct subset of lymphocytes that are characterized by the cell surface expression of a single invariant T-cell receptor (TCR) and Natural Killer (NK) cell markers. The activation of NKT cells is initiated by the engagement of the NKT receptor with glycolipid antigens presented by the molecule CD1d. Careful examination of Mekk1 kinase-deficient mice (Mekk1ΔKD) reveals multiple defects in NKT cells. Mekk1ΔKD NKT cells have deficient: MAPK signal transduction, proliferative expansion, apoptosis, and aberrant cytokine production. Mekk1ΔKD mice display elevated liver damage in a ConA-induced acute hepatitis liver injury model. My results demonstrate for the first time that MEKK1 is a critical MAPK Kinase kinase (MAP3K) signalling component for the regulation of NKT cells.
Control of T cell activation by Drak2 depends on protein kinase D and the interplay between calcium and reactive oxygen species (50.6)
Authors: Newton Ryan , Walsh Craig 
Published: January 2023

Abstract
Drak2 plays a negative regulatory role in acute T cell activation with direct consequences on proliferative capacity. The absence of Drak2 in T cells provokes responsiveness to sub-optimal stimuli, heightens calcium mobilization across a broad range of antigen receptor cross-linking, and increases production of Il-2, yet key survival factors are diminished within the context of Drak2 deficiency, ultimately limiting effector function. Increased co-stimulation, or transgenic expression of Bcl-xL, can rescue this proliferative defect. Thus, Drak2 provides a link between negative regulation of T cell activation and the requirement for adequate co-stimulation. The pathway through which this serine/threonine kinase functions has remained elusive. This work shows that Drak2 activity is dependent on protein kinase D (PKD), in a calcium-dependent manner, and that a PKD/Drak2 signaling axis serves to limit the influx of extracellular calcium following T cell activation. Furthermore, reactive oxygen species are required for Drak2 activity and are shown to play a key role in the initial phase of this negative feedback loop. Taken together, PKD and Drak2 are shown to limit the activation state of T cells through tuning those signals which initially converge to ultimately drive effector function.
PTPN4 and PTPN3 regulation of ITAM containing proteins (50.9)
Authors: Dougall David , van Oers Nicolai 
Published: January 2023

Abstract
The T cell receptor (TCR) uses a highly conserved ITAM signaling pathway to transduce intracellular signals following appropriate ligand binding. Activations of the ITAM signaling pathway is tightly controlled by diverse families of protein tyrosine kinases and protein tyrosine phosphatases. In earlier studies, we reported that two PTPases (PTPN3 and PTPN4) could directly dephosphorylate the tyrosine residues in the CD3 z ITAMs. However, T cells from both PTPN3 and PTPN4-deficient mice exhibit normal CD3 regulated ITAM signaling pathways. Since PTPN3 and PTPN4 have FERM domains that localize these enzymes to the cytoskeleton, it is possible that these PTPases target distinct proteins. Interestingly, moesin, ezrin, and dectin-1 are three cytoskeletal associated proteins that use ITAM sequences to transduce intracellular signals. To examine whether these distinct ITAM-containing proteins were dephosphorylated by PTPN3 or PTPN4, a combination of transient transfection assays and cell signaling studies in PTPase-deficient dendritic cells were performed. These experiments will reveal whether this PTPase-family dephosphorylates distinct ITAM-containing proteins.
Faithful epigenetic and transcriptional regulation of a transgenic human Th2 cytokine gene locus in the murine nuclear environment (51.1)
Authors: Pivniouk Vadim , Bailey Tom , Pivniouk Oksana , Kiesler Patricia , Wlasiuk Gabriela , Rosenbaum David , Kim Kenneth , Strempel Jannine , Daines Michael , Vercelli Donata 
Published: January 2023

Abstract
Dysregulated production of the Th2 cytokines IL4 and IL13 is a hallmark of allergy and asthma. Th2 cytokine gene locus regulation has been extensively studied in mice, much less in humans. We have generated mice carrying a 160 kb BAC transgene (BAC5) that encompasses human RAD50, IL13 and IL4 and includes all the elements known to be required for faithful regulation of mouse Th2 cytokine locus. BAC5 transgenic mice appeared normal, were born at the expected Mendelian ratio, and carried 1-15 intact copies of BAC5. In vitro polarized mouse transgenic CD4 Th2, but not Th1 cells expressed human IL13 and IL4, demonstrating tissue-specific regulation of human Th2 cytokines. Murine CD4 T cells expressed physiologic levels of human IL13 and IL4, particularly in a house dust mite immunization model. Human IL13 expression was transgene copy number-dependent, suggesting BAC5 includes the human Th2 locus control region. Notably, DNA methylation patterns at multiple DNase I hypersensitive sites in the Th2 locus of human CD4 T cells were highly comparable if not identical in transgenic CD4 T cells of equivalent differentiation stages. Our results support the notion that in homologous tissues, genetic sequence is largely responsible for directing transcriptional programs and chromatin modifications even in a heterologous nuclear environment. Moreover, our findings suggest BAC5 transgenic mice are a powerful tool to study human Th2 cytokine gene regulation and Th2-dependent events in vivo.
Characterization of a novel stimulation-dependent secondary promoter in the lymphotoxin alpha gene (51.12)
Authors: Posch Phillip , Yokley Brian 
Published: January 2023

Abstract
The lymphotoxin alpha (LTA) downstream region (DSR, +1 to +459) between the transcription and translation start sites contains polymorphisms that impact expression, yet the importance of the DSR to transcriptional regulation has not been examined. Similar regions in other genes have been shown to contain numerous regulatory elements and secondary promoter sites. We found that the LTA DSR is required for full activity of the LTA promoter in T and B cells as deletion of this region leads to a ≥55% reduction in luciferase expression under basal and stimulated conditions. Further, expression levels driven by the DSR alone were found to be comparable to that of the LTA regulatory region (-915 to +459) in basal and stimulated B and T cells. We hypothesized that the DSR contained a secondary promoter site that might consist of a putative Sp1 and/or TFII-I [initiator (Inr)-like element] binding sites located near the 3' end. Mutation of these sites resulted in significant reductions of DSR-mediated expression. ChIP assays showed that Sp1 and TFII-I, as well as RNA pol II, bind to this region of LTA in vivo. 5'RLM-RACE assays revealed LTA transcripts initiating from the Inr-like element when T cells were stimulated with human recombinant (hr) LT-α, hrTGF-β1 and hrKGF, but not when stimulated with PMA/ionomycin or left unstimulated. All transcripts were novel utilizing five novel start sites. These data show a complex stimulation-dependent pattern of LTA transcriptional regulation.
Alternatively spliced IL-4 protein is naturally secreted by T cells (51.2)
Authors: Bocharov Alexander , Lockatell Virginia , Lavania Sachin , Todd Nevins , Luzina Irina , Atamas Sergei 
Published: January 2023

Abstract
We have previously described an alternatively spliced isoform of IL-4 mRNA that omits exon 2 (IL-4δ2). We and others reported that the total levels of combined IL-4 + IL-4δ2 mRNAs and IL-4 / IL-4δ2 mRNA ratios change in patients with scleroderma, asthma, and various infections. Natural IL-4δ2 protein was not previously studied because most commercial anti-IL-4 antibodies (Ab) do not react with IL-4δ2. We established mammalian expression systems (HEK293 stably transfected or infected with recombinant adenoviral constructs) producing naturally processed (folded and glycosylated) human IL-4 or IL-4δ2. By screening a library of commercial anti-IL-4 Abs, we identified a selective Ab reacting exclusively with IL-4, and an indiscriminate Ab reacting equally with IL-4 and IL-4δ2. PBMC and T lymphocyte (&gt; 95% purity) cultures from 3 donors with allergic asthma and 3 controls were stimulated with PMA/ionomycin for 24 hrs. The supernates were immunodepleted of wild-type IL-4 using four cycles of depletion with the selective antibody. These immunodepleted supernates were negative in ELISAs with the selective anti-IL-4 Ab. However, the IL-4-depleted supernates from asthmatics, but not controls, were positive in ELISAs with the indiscriminate antibody that now detected only IL-4δ2. The levels of IL-4δ2 were 5 to 10 fold lower than the levels of IL-4 in the initial undepleted supernates. Thus, IL-4δ2 is naturally produced not only in the mRNA form, but also as a secreted protein.
Nourexin-4 A Novel Anti-inflammatory Therapy for Influenza Flu (52.1)
Authors: Elgebaly Salwa , Perez Daniel , Sullivan Kathleen , Whitaker Craig , Caspe Stephanie , Yi Qiao , Kreutzer Donald 
Published: January 2023

Abstract
Relatively little is known about the inflammatory mediators and mechanisms that drive the progression of influenza flu infection to cytokine storm, lung dysfunction, organ failure, and ultimately death. Vaccines and antiviral medications cannot control the excessive host inflammatory response. We demonstrated the rapid release of a potent inflammatory mediator, recently named Nourin, by local mammalian tissues in response to injury and infection. Nourin is a formyl peptide that acts through the formyl peptide receptor (FPR) on phagocytic leukocytes. As an “initial signal” in the “innate immunity”, Nourin stimulates leukocyte chemotaxis, induce acute and chronic inflammation, and stimulates the release of cytokine storm mediators from monocytes and neutrophils. Nourin detected in plasma samples from patients with severe influenza infection was much higher compared to moderate influenza. We then tested the Nourin antagonist Nourexin-4, as specific competitive antagonist of formyl peptides on phagocytic leukocytes FPR. Nourexin-4 completely blocked neutrophil chemotaxis induced by the standard formyl peptide f-MLF and the host-derived Nourin released by (1) cultured epithelial cells infected with the H1N1 influenza virus (PR8) (6-24 hours), (2) Nourin detected in the serum of mouse model of H1N1 influenza (6 hrs), and (3) Nourin detected in severe and moderate influenza patients plasma samples. Nourexin-4 can be used to control virus-induced inflammation and protect patients.
Route of immunization influences the induction of humoral, cellular and protective immunity by live attenuated measles vaccine in rhesus macaques (52.11)
Authors: Lin Wen-Hsuan , Pan Chien-Hsiung , Adams Robert , Ford Brandi , Laube Beth , Mikszta John , Sullivan Vince , Griffin Diane 
Published: January 2023

Abstract
Measles is a leading cause of vaccine-preventable mortality worldwide. To facilitate vaccine distribution, aerosol immunization has been proposed, but the immune response induced, and protection afforded, by live attenuated measles virus (MV) vaccine (LAV) given by the respiratory route have not been systematically studied. Rhesus macaques were immunized with liquid or powder LAV through a nebulizer, an endotracheal tube, or parenterally by intramuscular injection. The method of immunization significantly influenced the induction of the humoral and cellular immune responses to LAV, but the effect on the cellular immune response was not correlated with the effect on the humoral response. For instance, nebulizer immunization induced good T cell, but poor antibody responses. Intratracheal challenge with wild type MV showed that respiratory immunization did not fully prevent infection. However, animals primed with nebulizer LAV showed accelerated viral clearance from both blood and respiratory tract that coincided with faster and higher levels of MV-specific recall CD4+/CD8+ T-cell responses compared to controls. Therefore, the route of immunization influences the induction of humoral, cellular and protective immune responses to measles vaccine. Protection correlates with neutralizing antibody and not with T-cell responses. Although, memory T cells alone are not sufficient for preventing MV infection, a robust recall T cell response does accelerate viral clearance.
Construction of multimerized scFv that neutralize Bovine Herpes Virus-1 (52.12)
Authors: Pasman Yfke , Nagy Eva , Kaushik Azad 
Published: January 2023

Abstract
Our laboratory earlier constructed functional antibody fragments, scFv (single chain fragment variable), with an 18 amino acid linker, against bovine herpes virus-1 (BoHV-1) (Koti et al. 2010. Molec. Immunol. in press). With an objective to enhance scFv avidity, the scFv in Vl-linker-VH configuration was constructed with a short flexible amino acid linker. An overlap PCR product comprising Vl-linker-VH was cloned into pPICZα vector and expressed in Pichia pastoris, under the influence of inducible AOX1 promoter. Affinity purified scFvs were analyzed by Western blot and mass spectrometry that confirmed the expected molecular weight of the scFv. Further, the multimerized scFvs, both purified and unpurified, neutralized BoHV-1 infection of MDBK cells in vitro. Taken together with previous studies, the scFvs with varying linker size are capable of effectively neutralizing BoHV-1. The scFv against BoHV-1 will provide protection at respriratory mucosal surfaces and prevent infection. In conclusion, scFvs both in multimerized and non-multimerized form provide a useful diagnostic and therapeutic agent against an important cattle pathogen. (Supported by NSERC Canada)
Totally synthetic nanoparticle vaccines produced via layer-by-layer assembly of polypeptide films induce protective T-cell responses (52.14)
Authors: Powell Thomas , Palath Naveen , DeRome Mary , Tang Jie , Jacobs Andrea , Boyd James 
Published: January 2023

Abstract
Nanoparticle vaccines were synthesized using layer-by-layer (LbL) deposition of oppositely-charged polypeptides to build up an artificial film on CaCO3 nanocores. Designed peptides (DP) containing antigenic epitopes of interest were incorporated into the layers of the film, yielding nanoparticles that delivered the DP payload to dendritic cells via phagocytosis. LbL nanoparticles induced DC maturation without triggering secretion of inflammatory cytokines including TNFα and IL-6. LbL nanoparticle delivery of DP to DC resulted in potent cross-presentation to CD8+ T-cells and more efficient presentation to CD4+ T-cells compared to presentation of soluble peptide. Mice immunized with a single dose of LbL nanoparticles without adjuvant mounted vigorous T-cell responses detectable by IFNγ and IL-4 ELISPOT and in vivo CTL assay. Mice immunized with nanoparticles containing ovalbumin-derived DP were protected from challenge with Listeria monocytogenes ectopically expressing ovalbumin. Similar to the in vitro DC maturation results, immunization with LbL nanoparticles did not induce inflammatory symptoms locally (injection site reaction) or systemically (serum cytokine spike). The potency and efficacy of polypeptide LbL nanoparticles administered in aqueous suspension without adjuvant or other formulation additive, and the absence of overt inflammatory responses, suggest that this strategy may be useful in producing novel vaccines against multiple diseases.
Mechanisms of post-exposure protection mediated by virus-like particles against Ebola virus infection (52.18)
Authors: Bradfute Steven , Tailor Prafullakumar , Anthony Scott , Ozato Keiko , Bavari Sina 
Published: January 2023

Abstract
Ebola virus causes hemorrhagic fever that is characterized by high viral titers, coagulopathy, and high lethality rates. Our lab has developed a virus-like particle (VLP) vaccine that is effective in mouse, guinea pig, and non-human primate models when given as a pre-exposure vaccine. In this work, we show that VLPs protect Ebola-infected mice when given 24 hours post-infection. We used this platform to analyze mechanisms of immunity for post-exposure protection against Ebola virus. Analysis of serum revealed a decrease in many cytokine levels in mice given VLPs post-exposure compared to infected, untreated mice. VLP-mediated post-exposure protection required B cells, perforin, STAT-1 signaling, and either CD4+ or CD8+ T cells. However, NK cells, IFN-gamma, and TNF-alpha were not required for this protection. Interestingly, transient depletion of macrophages or dendritic cells abrogated VLP-mediated post-exposure protection. Additional studies are underway to further characterize the mechanisms of immunity in this setting.
Preclinical evaluation of a 15-valent pneumococcal polysaccharide-protein conjugate vaccine in infant rhesus monkeys (52.20)
Authors: Skinner Julie , Indrawati Lani , Winters Michael , MacNair John , Manger Walter , Pujar Hari , Blue Jeffrey , Antonello Joseph , Heinrichs Jon , Caulfield Michael 
Published: January 2023

Abstract
The incidence of invasive pneumococcal disease (IPD) caused by the 91 serotypes of Streptococcus pneumoniae (PN) varies geographically and temporally as a result of changing epidemiology and vaccination. Prevnar® (Pfizer), the first licensed pneumococcal conjugate vaccine (PCV), comprises polysaccharides (PS) from 7 serotypes conjugated to the mutant diphtheria toxin carrier protein, CRM197. In the United States, this vaccine has been highly efficacious in reducing the incidence of IPD caused by vaccine serotypes, however, the incidence of non-vaccine serotypes (e.g., 19A, 22F, and 33F) has increased resulting in the need for vaccines with higher valencies. Indeed, 10- and 13-valent PCVs have recently been licensed. For expanded coverage, we have developed a 15-valent PCV containing PS from serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F conjugated to CRM197 and formulated on aluminum phosphate adjuvant. The vaccine was tested in infant rhesus monkeys since they, like human infants, do not respond to unconjugated PN PS. Infant (2-3 month old) rhesus monkeys were vaccinated 3 times with PCV-15 or Prevnar® at 2 month intervals, and serotype-specific IgG antibodies were measured using a multiarray electrochemiluminescence (ECL) assay. The results indicate that antibody responses to PCV-15 and Prevnar® were comparable for the 7 common serotypes and that post-vaccination responses to PCV-15 were &gt; 10-fold higher than baseline for the 8 added serotypes.
Alphavirus particle filovirus vaccine activation of Dendritic cells (52.7)
Authors: Pisarcik Sarah , Herbert Andrew , Olinger Gene 
Published: January 2023

Abstract
Filoviruses, Marburg (MARV) and Ebola (EBOV), are among the deadliest etiological agents causing hemorrhagic fevers. Although little is known about the pathogenesis or what immunological responses are required to protect against filoviruses, a promising recombinant RNA replicon vaccine has been developed consisting of an attenuated alphavirus, Venezuelan equine encephalitis virus (VEE), to express MARV and EBOV viral genes such as the virus glycoprotein (GP). While numerous in vivo studies have demonstrated the protective efficacy of filovirus GP expressing VEE replicon particles (GP-VRP) in mice, guinea pigs, and non-human primates, when challenged with homologous virus, little has been done to understand the means by which these VRP filovirus vaccines initiate and establish immunity. In this study, we sought to dissect the protective mechanism of VRPs by evaluating the GP-VRPs effects on dendritic cells (DCs) by several characterization assays. Immunofluorescence assay (IFA), flow cytometry, and cytokine detection assays were used to assess maturation, co-stimulatory molecule expression, and cytokine expression profiles of GP-VRP-treated DCs. These findings will aid in deciphering early innate immune responses initiated by GP-VRP vaccination that are necessary for eliciting protective immunity against filovirus infection.
Placental extract suppresses mouse models of autoimmunity (83.4)
Authors: Landek-Salgado Melissa , Kimura Hiroaki , Rose Noel , Caturegli Patrizio 
Published: January 2023

Abstract
Pregnancy is known to modulate many autoimmune diseases. For example, autoimmune thyroiditis improves during pregnancy, and autoimmune hypophysitis presents during this period. The mechanisms underlying the effect of pregnancy on autoimmunity remain unknown. To determine whether placental proteins influence autoimmunity, we used a mouse model of experimental autoimmune hypophysitis (EAH). Here we report that co-immunization of mice with placental extract and pituitary extract, significantly decreased EAH incidence, severity, and antibody titers. Disease suppression was specific to the placenta; liver or muscle extracts were incapable of suppressing disease. Disease suppression, however, was not specific to EAH: co-immunization of placental extract with thyroglobulin, similarly decreased antibody titers. Immunization with pituitary extract during early or late pregnancy and administration of non-emulsified placental extracts were ineffective in suppressing EAH. Suppression of EAH was only observed when the placental extract was at the same location as the immunogen. Placental extracts were found to contain high levels of soluble TNF receptor one (sTNFR1). Furthermore, a placental extract deficient in TNFR1 was incapable of suppressing EAH, suggesting that sTNFR1 may be responsible for the placental-mediated suppression of EAH. These results suggest that a placental component, possibly sTNFR1, is capable of dampening the immune response to a co-injected immunogen.