bluuebunny/crossref_metadata_2025_split huggingface dataset:
Age influences the lymphocyte response to the ellagitannin oenothein B and acai polysaccharides. (VET2P.1040)
Authors: Ramstead Andrew , Schepetkin Igor , Quinn Mark , Jutila Mark 
Published: January 2023

Abstract
The effects of nutritional supplements on immunity are still not well understood. For example, although aging affects immune cell responsiveness, it is not well known if immune stimulation by some nutritional supplements is also affected by aging. We have previously shown that the ellagitannin oenothein B and Acai berry polysaccharides (AcaiPS) enhance IFNγ production by both bovine and human NK cells and T cells. We wanted to examine the effect age had on this activity. Studies comparing immune responses in immature individuals to those in adults are difficult to perform in humans and mice. As such, we utilized the bovine system to examine the responsiveness of T cells and NK cells from newborn calves, older calves, and adult cows to oenothein B and AcaiPS in vitro. Oenothein B induced greater IFNγ production by T cells from adults compared to calves. Within the responsive T cell population, we found that CD45RO+ T cells expressed more IFNγ in response to oenothein B than CD45RO- T cells. Similar responses were seen in a limited analysis with human cells. In contrast, AcaiPS, both alone and in combination with IL-18, induced greater IFNγ production by NK cells from calves, especially newborn calves, compared to adults. Our data suggest that immune stimulation of T cells and NK cells by oenothein B and AcaiPS is influenced by age. It also suggests that the bovine model may be very useful in examining the effect of age on immune stimulation by nutritional supplements.
Anti-fungal immune responses to Pseudogymnoasces destructans in bats affected by white-nose syndrome (VET2P.1041)
Authors: Field Kenneth , Reeder Sophia , Rogers Elizabeth , James McMichael , Sigler Lauren , Vodzak Megan , Moore Marianne , Johnson Joseph , Reeder DeeAnn 
Published: January 2023

Abstract
White-nose syndrome is a devastating disease affecting bats in North America. It is caused by Pseudogymnoasces destructans, a fungal pathogen that colonizes the muzzles and wings of hibernating bats. Our study focuses on the relative differences between the immune responses of various bat species to this pathogen. The strength and type of immune response triggered by an antigen can be measured by determining the gene expression profile of cytokines. We have developed a quantitative PCR panel for bat cytokine genes in order to measure the immune responses involved in susceptibility to white-nose syndrome. We also used a cytokine ELISA to determine serum cytokine levels. In addition, we analyzed the neutrophil:lymphocyte ratios from blood smears collected both during hibernation and post hibernation, as another measure of immune response over time. By comparing the relative expression of certain cytokine genes, we can determine which subsets of T cells are mediating the immune response to the fungal pathogen. Our research will shed light on the mechanisms by which some bat species have a higher survival rate, and this knowledge can be used to further develop strategies for protecting North America’s bat population.
Mucosal correlates of cross-protection for live-attenuated influenza virus vaccines in pigs (VET2P.1031)
Authors: Hughes Holly , Vincent Amy , Loving Crystal 
Published: January 2023

Abstract
Controlling swine influenza A virus (IAV) has become increasingly difficult with the emergence of novel reassorted strains and introduction of human seasonal IAV into pigs. In North American swine there are 6 antigenically distinct H1 subtypes currently circulating. Live-attenuated influenza vaccines (LAIV) provide broader cross-protection than whole-inactivated virus (WIV) vaccine making LAIV a candidate for next-generation swine IAV vaccines. However, there is no standardized assay to predict cross-protection following LAIV vaccination. Hemagglutination-inhibiting (HI) immunoglobulin (Ig) in serum has long been the gold standard correlate of protection following WIV vaccination; however, LAIV does not elicit a robust serum HI Ig titer. Oral fluids (OF) have become a rapidly developing diagnostic specimen for a variety of animal pathogens. We evaluated nasal wash (NW) and OF samples for measuring cross-reactive Ig responses to a panel of H1 viruses following vaccination. Both NW and OF had detectable levels of IAV-specific Ig when measured by ELISA. NW from LAIV vaccinated pigs had significant cross-reactive IgA titers, while NW from WIV vaccinated pigs did not have detectable IgA. Samples of OF from either LAIV or WIV vaccinated pigs had high titers of cross-reactive IAV-specific IgG and measurable IgA. OF samples also displayed IAV specific neutralization. These data suggest OF can serve as a sample for evaluating LAIV immunogenicity and predicting cross-protection.
Progress in using coral as a model to study intrinsic and innate immune responses to herpes-like viruses (VET2P.1032)
Authors: Garrison Keith , Hernandez Austin 
Published: January 2023

Abstract
Corals have been shown to produce herpes-like viruses when triggered by abiotic stressors. In addition, new associations are being discovered between known coral diseases and viral pathogens. We intend to use coral as a model species to study invertebrate anti-viral immune responses in organisms critical to the resilience of the coral reef ecosystem. Our long-term goals include identifying possible immune genes in corals and measuring changes in gene expression in response to virus production. Fragments of Acropora were propagated in controlled aquarium systems mimicking fundamental properties of shallow reef with live rock, live sand, and artificial seawater. We acclimatized corals by providing sufficient photosynthetic active radiation, feeding, and periodic replenishment of macro- and microelements. RNA extractions were performed on coral specimens to synthesize cDNA for qPCR. Known human innate immune pathways such as STING, cGAS, Factor X, and NF-kB are under investigation using primers targeting the coral orthologs of critical human genes in these pathways. We are optimizing qPCR as a tool to amplify and quantify target genes from cDNA. We are also investigating western blot anaylsis of NF-kB to assess levels of immune activation in coral cells. Eventually, we hope to develop minimally-invasive tests to provide leading indicators of coral stress to help identify and manage threats to reef ecosystem health.
Distinct and overlapping roles for cell intrinsic IL-6 and IL-27 signaling in promoting virus specific CD4+ T cell numbers and function during acute and chronic viral infections (VIR7P.1055)
Authors: Harker James , Dolgoter Aleksandr , Lewis Gavin , Zuniga Elina 
Published: January 2023

Abstract
Chronic viral infections such as human immunodeficiency virus 1 and hepatitis C virus represent a significant global health burden. Virus specific CD4+ T cell number and function are critical in determining the outcome of these chronic viral infections, but little is known about the signals that can promote their survival or helper functions in this environment. By studying both acute and chronic variants of lymphocytic choriomeningitis virus infection of mice we found that the IL-6 cytokine family signal transduction molecule gp130 expression on CD4+ T cells was essential for long term viral control in chronic, but not acute infection. It was critical for virus specific CD4+ T cell survival, differentiation and the production of IL-21, factors which are critical in the control of chronic infections. Dissection of the individual IL-6 family of cytokines revealed that IL-6 was critical for the upregulation of the transcriptional regulator Bcl6 and subsequent escalation in T follicular helper cell responses and development of antiviral antibody. Contrastingly IL-27 was uniquely required in the survival of these virus specific CD4+ T cells at late stages of chronic infection. Use of Il6ra-/-;Il27ra-/- ¬mice showed however that IL-6 and IL-27 had an overlapping role in the production of IL-21. Our data situates the IL-6 cytokine family as central orchestrators of anti-viral CD4+ T cell responses and subsequent immune responses vital during established chronic viral infections.
High levels of interleukin-21(IL-21) are produced during primary dengue infections. (VIR8P.1067)
Authors: Maldonado Rentería Mathywz , Sánchez Vargas Luis , Izaguirre Irma , Hernández Flores Karina , Vivanco Cid Héctor 
Published: January 2023

Abstract
Introduction: IL-21 is produced by CD4 T cells, mainly follicular T cells and exerts multiple and pleiotropic effects on innate and adaptive immune as well as on non-immune cells. IL-21 is involved in various viral infections but has not been described in dengue virus (DENV) infection context. Objective: The aim of this study was to investigate the IL-21 level production during the clinical course of primary DENV infections. Material and Methods: We collected 122 serum samples of primary DENV-infected patients at different days after the onset of symptoms and classified according to their phase of disease (acute vs. convalescent phases), and the clinical severity (dengue fever vs. dengue hemorrhagic fever). IL-21 levels were measured using a quantitative capture ELISA. Results: IL-21 levels were significantly elevated in the disease group compared with the control group (P &lt; 0.0001). IL-21 was detected in primary DENV infections with a significantly higher concentration in the convalescent phase. Conclusions: Our results show for the first time the involvement of IL-21 during the clinical course of DENV infections. We speculate that IL-21 may play a protective role in primary infections.
Peptide presentation by different HLA-Class I molecules during viral infection (APP2P.106)
Authors: Kaabinejadian Saghar , Vernon Stephen , McMurtrey Curtis , Mojsilovic Danijela , Bardet Wilfried , Schafer Fredda , Jackson Kenneth , Buchli Rico , Piazza Paolo , Hildebrand William 
Published: January 2023

Abstract
Class I Human Leukocyte Antigens reveal intracellular viruses to the immune system by presenting viral epitopes at the cell surface. In this study we hypothesized that during infection particular HLA class I alleles consistently present more virus-derived peptide ligands to virus-specific CD8+ T cells than do other HLA class I. To test this hypothesis, we infected cells with West Nile virus (WNV) and directly identified and enumerated the virus specific peptides presented by various HLA class I molecules. Cells expressing a number of different HLA-A and HLA-B class I were cultured in bioreactors and infected with WNV. HLA/peptide complexes were harvested from infected and uninfected cells, peptides were eluted from affinity-purified HLA, comparatively mapped by mass spectroscopy, and sequenced. Twenty-four peptides, 20 eluted from HLA-A complexes (HLA-A*02:01, A*01:01, A*11:01 and A*24:02) and 4 from HLA-B complexes (HLA-B*27:05, B*07:02 and B*35:01), were identified as unique to infected cells. Ligands represented different parts of the viral polyprotein demonstrating that peptides sampled by class I HLA are distributed widely throughout the WNV proteome. These data indicate that, in WNV-infected cells, HLA-A present more virus-specific peptides than do HLA-B suggesting a potentially different role for HLA class I loci in developing immune responses that control WNV infection whereby HLA-A is responsible for diverse reactivity and HLA-B leads to more focused immunity.
Detection and validation of HLA Class I presented viral epitopes using T cell receptor mimics (APP2P.107)
Authors: Kaabinejadian Saghar , McMurtrey Curtis , Kim Sojung , Jain Rinki , Bardet Wilfried , Schafer Fredda , Diamond Michael , Hansen Ted , Weidanz Jon , Hildebrand William 
Published: January 2023

Abstract
Class I Human Leukocyte Antigens distinguish healthy cells from infected cells by presenting peptides at the cell surface. Viral epitopes confirmed as unique to infected cells can lead to successful development of vaccines and therapeutics, and a tool to validate viral epitope presentation on a variety of cell lineages is essential. Here, comparative mass spectrometry shows that the West Nile virus peptide epitopes SVGGVFTSV and ILRNPGYAL are presented by HLA-A*02:01 and HLA-B*07:02 of infected cells. To generate monoclonal antibodies against these HLA/WNV peptide epitopes, mice were immunized with peptide/HLA complexes, splenocytes were fused to myeloma cells, and single clones were picked and grown. Hybridoma supernatants were screened for recognition of the appropriate HLA/WNV peptide complex on peptide-pulsed cells with irrelevant HLA/peptides acting as a negative control. The T cell receptor mimic (TCRm) monoclonal antibodies RL15A and RL29A were found to be specific for A*02:01/SVG9 and B*07:02/ILR9, respectively. The RL15A and RL29A T cell receptor mimic mAb were then used to track viral epitope presentation on WNV infected cell lines and primary cells. In summary, we demonstrate the implementation of a mass spectrometry system for the direct discovery of class I HLA-presented viral epitopes from infected cells followed by the complementary use of TCRm mAb as a companion diagnostic for tracking epitope presentation during the course of a viral infection.
HIV-1 NEF ligands predominate in the HLA class I of infected CD4+ T cells (APP2P.108)
Authors: Yaciuk Jane , Smith S. , Skaley Matthew , McMurtrey Curtis , Bardet Wilfried , Schafer Fredda , Mojsilovic Danijela , Stewart Christopher , Cate Steven , Jackson Kenneth , Weidanz Jon , Olvera van der Stoep Alex , Brander Christian , West John , Hildebrand William 
Published: January 2023

Abstract
Class I HLA reveal virus-derived ligands to cytotoxic T lymphocytes (CTL) whose function is to eliminate infected cells. The Los Alamos National Laboratory (LANL) database reports numerous HIV-1 CTL epitopes, none of which have elicited protective immunity in vaccine testing. The goal of this study was to determine the number and nature of HIV-1 ligands available for CTL targeting through presentation by the HLA class I of virus-infected cells. Class I presented peptides were recovered from immunoaffinity purified HLA-A*11:01 gathered from HIV-1 (NL4-3)-infected human CD4+ SUP-T1 T cells. These peptides were fractionated by RP-HPLC and mapped by tandem mass spectrometry (MS/MS). Seven HIV-derived peptides were confirmed by MS/MS as unique to infected cells. Twelve LANL HIV epitopes, including the well-studied Gag p24 epitope ACQGVGGPGHK (AK11), were clearly absent from the HLA-A*11:01 of infected cells. Of the 7 HIV-1 ligands found to be presented by A*11:01, 4 were derived from Nef, 2 from Gag, and 1 from Pol. When these 7 ligands were tested for T cell reactivity in a gamma interferon ELISPOT assay using PBMC from HIV-1 infected individuals (including elite controllers), the Nef-derived peptides were found to be highly reactive. These data demonstrate that the class I HLA of HIV-1 infected CD4+ T cells present a handful of viral peptide ligands for recognition by CTL and that reported HIV-1 CTL epitopes might not be presented by the HLA class I of infected cells.
LC3-associated phagocytosis mediates IFN-alpha secretion in response to DNA-immune complexes. (BA3P.207)
Authors: Mehta Payal , Henault Jill , Martinez Jennifer , Riggs Jeffery , Tian Jane , Clarke Lorraine , Sasai Miwa , Latz Eicke , Brinkmann Melanie , Iwasaki Akiko , Coyle Anthony , Kolbeck Roland , Green Douglas , Sanjuan Miguel 
Published: January 2023

Abstract
TLR9 compartmentalization is largely responsible for discriminating self from pathogenic DNA. However, association of host DNA with autoantibodies activates TLR9, inducing the secretion of large amounts of type I IFNs in plasmacytoid dendritic cells. Here, we show that TLR9-mediated production of IFN-α depends on the convergence of the phagocytic and autophagic pathways in response to DNA-containing immune complexes, a process called LC3-associated phagocytosis (LAP). LAP is required for TLR9 trafficking into a specialized IRF7-signaling compartment (ISC) by a mechanism that involves ATG7 but not the conventional autophagic pre-initiation complex. Progression to this compartment did not require AP-3, a protein that mediates TLR9 trafficking and IFN-α secretion in response to CpG oligonucleotides. Our findings unveil a new role for autophagy in inflammatory responses, and provide one of the mechanisms by which anti-DNA autoantibodies, such as those found in several autoimmune disorders, bypass the controls that normally restrict TLR9 activation to pathogenic DNA. Further, we have established phagosomal enrichment assays that will allow us to characterize precise nature ISC as well as the role of LAP in ISC regulation .
The arginine deminase PAD2 is activated by ATP-induced P2X7R signaling in mast cells and modulates the disease course in an inflammatory arthritis model (BA3P.208)
Authors: Mowen Kerri , Guimond David , McKenney Katherine , Arandjelovic Sanja 
Published: January 2023

Abstract
Rheumatoid arthritis is a chronic, systemic inflammatory disease. Citrulline-containing proteins are generated through posttranslational modification of arginine residues in a reaction catalyzed by the Ca2+-dependent peptidyl arginine deiminases or PAD enzymes. Plasma and synovial biopsy specimens from RA patients contain high levels of citrullinated proteins. PAD2 is highly expressed in synovial tissue of RA patients, in close association with citrullinated protein deposits, and its expression correlates with inflammation intensity. The process by which PAD2 is activated and contributes to protein citrullination during RA is unknown. We have identified mast cells as a major source of the PAD2 enzyme. Activation of the P2X7 purinergic receptor by the inflammatory “danger” signal ATP induces PAD2 activity and robust protein citrullination in mast cells. P2X7-mediated activation of PAD2 is sensitive to p38 MAPK and protein kinase C inhibitors, and PAD2 regulates the expression of the TNFR2, Adamts-9, and Rab6b transcripts in mast cells. The PAD2 enzyme and its citrullinated substrate proteins are released from mast cells on activation with ATP. Importantly, mast cells have been shown to play a key role in the development of RA. Our results suggest that P2X7 activation of mast cells may play a role in inflammation by providing PAD2 and PAD2 substrates access to the extracellular space.
Diferuloylmethane suppresses the inflammatory response that mediates type 1 diabetes development (BA3P.133)
Authors: Alard Pascale , Lee Tricia , Rahmouni Faiza , Kosiewicz Michele 
Published: January 2023

Abstract
Because of their various immune defects including the excess production of proinflammatory cytokines by dendritic cells (DC), non-obese diabetic (NOD) mice provide a powerful model to understand type 1 diabetes. Diferuloylmethane (cucurmin), a natural phenol chemical isolated from the plant Curcuma longa (turmeric), is believed to be anti-inflammatory, and has been shown to ameliorate autoimmune diseases such as multiple sclerosis, rheumatoid arthritis, psoriasis, and inflammatory bowel disease in human and/or animal models. We have, therefore, examined the effects of diferuloylmethane on the inflammatory response leading to type I diabetes. We have found that DC from NOD mice produce decreased pro-inflammatory cytokines, including IL-12, following treatment with diferuloylmethane in vitro. More importantly, diferuloylmethane-treated NOD DC induce fewer IFNγ-producing T cells, shown to be responsible for beta islet destruction. Cell viability was not affected by treatment with diferuloylmethane as shown by 7AAD labeling. Finally, T cells from the spleens of NOD mice treated in vivo with diferuloylmethane for several weeks produced significantly less IFNγ. Overall, our data show that diferuloylmethane suppresses in vitro and in vivo the differentiation of inflammatory cells involved in type 1 diabetes development. Diferuloylmethane may, therefore, have therapeutic potentials for preventing the development of type 1 diabetes.
Central nervous system-specific expression of C-reactive protein exacerbates experimental autoimmune encephalomyelitis. (BA3P.202)
Authors: Wright Tyler , McCrory Mark , Morgan Todd , Bali Namrata , Hou Xiaogang , Finch Caleb , Szalai Alexander 
Published: January 2023

Abstract
Long considered merely a marker of inflammation, new evidence indicates that C-reactive protein (CRP) may also affect the maintenance of health and propagation of disease. Our lab previously showed that human CRP, hepatically synthesized by human CRP transgenic mice (CRPtg), delays onset and reduces severity of Experimental Autoimmune Encephalomyelitis (EAE; a model of Multiple Sclerosis). Though no causal data are available for humans, research suggests that unlike CRPtg, expression/deposition of CRP in the CNS associates with disease, rather than protection. Though human CRP expression in both man and CRPtg mouse is largely hepatic, studies have shown that it is also expressed by multiple cells of the CNS, particularly during neuroinflammation. To test CNS-expressed CRP’s impact on EAE, we made transgenic mice wherein the neuron-specific Ca2+/calmodulin-dependent protein kinase IIα (CaMKIIα) promoter drives human CRP expression. We confirmed that human CRP expression is restricted to the CNS in these neuronal CRPtg (nCRPtg) mice. In contrast to CRPtg, CNS-specific expression of CRP neither delays EAE onset nor improves its symptoms. As in humans, CNS-expressed CRP associates with more severe disease. In summary, observations made with CRPtg vs. nCRPtg reveal a previously unappreciated site-specificity to the impact of CRP on the CNS. The possibility that CRP mediates beneficial vs. detrimental actions depending on the local cellular environment will be investigated.
Transcription factor Fli-1 directly regulates the expression of IL-6 (BA2P.114)
Authors: Zhang Xian , Sato Shuzo , Lennard Richard Mara 
Published: January 2023

Abstract
Fli-1, a member of the Ets family of transcription factors, is highly expressed in immune cells and endothelial cells and is implicated in the development of systemic lupus erythematosus (SLE) in both human patients and mouse models. Increased expression of IL-6 is also involved in the SLE and other autoimmune disease development. We examined the role of Fli-1 on production of IL-6 in this study. We found expression of IL-6 was significantly reduced in sera from Fli-1 heterozygous knockout MRL/lpr mice (Fli1+/-; Fli-1 homozygous knockout is embryonic lethal), a murine model of lupus, compared to wild-type littermates. The production of IL-6 were significantly decreased in endothelial cells transfected with specific Fli-1 siRNA compared to cells transfected with negative control siRNA after LPS stimulation. We demonstrated that Fli-1 directly binds to the IL-6 promoter by Chromatin Immunoprecipitation (ChIP) assay and transient transfection assays show that Fli-1 drives transcription from the IL-6 promoter in a dose-dependent manner. Our data indicate that transcription factor Fli-1 binds to the promoter of IL-6 and directly regulates the expression of IL-6.