bluuebunny/crossref_metadata_2025_split huggingface dataset:
A single immunization with optimized DNA vaccines protects against lethal Ebola virus challenge in mice (VAC8P.1057)
Authors: Patel Ami , Scott Veronica , Wong Gary , Reuschel Emmal , Villarreal Daniel , Muthumani Karuppiah , Shedlock Devon , Yan Jian , Tierney Kevin , Sardesai Niranjan , Kobinger Gary , Weiner David 
Published: January 2023

Abstract
Ebola is a hemorrhagic fever virus that is responsible for the severe, ongoing outbreak in West Africa that has infected &gt;20000 people, exhibiting &gt;37% mortality. Infection control has been complicated by a lack of approved vaccines and therapies and recent figures suggest that the outbreak is more severe than previously thought. We generated 3 DNA vaccines expressing Zaire ebolavirus (EBOV) glycoprotein (GP): 2 vaccines designed on consensus alignments of EBOV GPs (1976-2014) and a 3rd matched construct to a 2014 Guinea strain. BALB/c mice received 40ug of each vaccine, delivered by IM injection followed by electroporation. Strong T cell responses were detected for all 3 vaccines, including GP-specific CD4+ and CD8+ T cells. To broaden efficacy against emerging EBOV strains, we also co-administered DNA vaccines in 2 or 3 construct formulations. Groups of 10 BALB/c mice were challenged with a lethal dose of a heterologous mouse-adapted Mayinga 1976 EBOV strain. The two consensus DNA vaccines, and combinations of 2 and 3 vaccines were 100% protective in mice. The matched vaccine afforded 90% protection. Total GP-specific IgG antibody levels were high in all surviving animals and low in unprotected animals suggesting that titers may indicate therapeutic efficacy. All three DNA vaccines are immunogenic and no antigen interference was observed. These experiments in mice support further studies in non-human primates towards continued development of this potential vaccine platform.
Immunization with superantigen-derived oligopeptides protect against S. aureus bacteremia (VAC8P.1058)
Authors: Karauzum Hatice , Datta Sandip 
Published: January 2023

Abstract
Staphylococcal superantigens are exotoxins that play a major role in disease pathogenesis. Using a T cell epitope prediction tool from the Immune Epitope Database Analysis Resource (IEDB) site, sequences within staphylococcal enterotoxin A and B (SEA, SEB) and Toxic Shock Syndrome Toxin with high affinity towards H2-Iab were identified and oligopeptides SEA_aa116-128, SEB_aa16-28 and TSST-1_aa212-224 were generated. C57BL/6 mice were immunized with a cocktail of oligopeptides or whole toxins in Al(OH)3. Serum samples from peptide-immunized mice tested positive towards individual peptides and whole toxins, however, samples from mice immunized with whole toxins did not bind to any of the peptides. To determine peptide-specific T cell response primed CD4 T cells isolated from spleens and total LNs of immunized mice were co-cultured with peptide-pulsed BMDCs and cytokine response was evaluated. Only cells of peptide-immunized mice resulted in elevated levels of IL-4, IL-5, IL-10, IL-13, GM-CSF, IFNg and TNFa, while cells obtained from whole toxin-immunized mice did not differ from baseline levels. To evaluate protective efficacy of peptides against toxin expressing strains, immunized mice were infected intravenously with MNHOCH (SEB+) or Newman (SEA+) and monitored up to 13 days. Mice that were infected with MNHOCH showed significantly extended survival compared to non-immunized mice (P=0.03). No significant difference in survival was observed when infected with strain Newman.
The yeast quorum sensing molecule, farnesol, acts as an adjuvant to promote innate inflammatory responses (VAC8P.1060)
Authors: Brown Deborah , Hargarten Jessica , Atkin Audrey 
Published: January 2023

Abstract
Candida albicans is a polymorphic fungus that can cause systemic disease in immunocompromised and transplant patients, but is maintained as a commensal in healthy adults mainly through the actions of innate immunity. At low cell density, C. albicans secretes the quorum-sensing molecule, farnesol (FOH) which acts as a virulence factor. Our previous work indicates that FOH alone promotes chemotaxis of macrophages (MΦ) and synergizes with the TLR-2 agonist zymosan to promote MΦ cytokine secretion in vitro. We next determined whether FOH could act similarly to promote inflammatory responses in vivo. After intra-peritoneal injection, FOH increased the numbers of MΦ and neutrophils in the peritoneal cavity compared to untreated or thioglycollate treated mice. FOH treatment increased IL-6 and CCL2 mRNA in peritoneal exudate cells consistent with MΦ activation. In addition, cell surface expression of MHC class II and co-stimulatory molecules on dendritic cells in the peritoneum were higher in FOH injected mice compared to controls, suggesting FOH may prime antigen presenting cells for T cell activation. Currently we are investigating whether FOH can act as an adjuvant to promote antigen specific, cell mediated or antibody mediated immunity or a combination of both types. Defining the immunostimulatory properties of FOH, a potentially novel adjuvant, will improve strategies for vaccine design that promotes robust immunity against a variety of infectious diseases.
Use of heterodimeric IL-15 in immune therapeutic regimens against HIV-1 or SIV infection (VAC8P.1048)
Authors: Pavlakis George , Bergamaschi Cristina , Valentin Antonio , Kulkarni Viraj , Jalah Rashmi , Rosati Margherita , Alicea Candido , Sowder Raymond , Chertova Elena , Felber Barbara 
Published: January 2023

Abstract
Interleukin-15 (IL-15) promotes the growth of NK and memory CD8 T cells, which makes it an attractive candidate in immune therapeutic strategies to eradicate HIV-1 infection. We have shown that the relevant IL-15 molecule in vivo is the heterodimer (hetIL-15) composed of IL-15 chain and the so-named IL-15 Receptor alpha (IL-15Ra) chain. We developed expression vectors producing high levels of bioactive hetIL-15. We have also developed stable cell lines overproducing IL-15 heterodimers. SIV-infected and uninfected rhesus macaques were treated with these DNA plasmids by electroporation. In addition, SHIV-infected macaques received repeated subcutaneous injections of purified hetIL-15 protein or placebo. Electroporation of plasmid DNA resulted in increased levels of circulating IL-15, which typically peaked between day 4 and 5 in all the animals. Similarly, treatment with hetIL-15 protein gave sustained high cytokine plasma levels without any significant changes in plasma VL or any other adverse effects. In both studies, increased levels of IL-15 resulted in early decrease in the percentage of NK cells, probably linked to their mobilization out of the bloodstream, and preferential accumulation of CD8+ T lymphocytes with effector memory phenotype. In conclusion, the combination of CD8 effector cells expansion and lack of viral activation makes IL-15 an ideal component in immune therapeutic strategies for the treatment and potential eradication of HIV-1 infection.
Novel vaccination of supplementing live attenuated influenza vaccine with M2 ectodomain tandem repeat to confer universal protection (VAC8P.1051)
Authors: Lee Young-tae , Kim Min-Chul , Ko Eun-Ju , Lee Yu-Na , Kim Ki-Hye , Kwon Young-Man , Hwang Hye Suk , Lee Youri , Jung Yu-Jin , Kang Sang 
Published: January 2023

Abstract
Current influenza vaccines including live attenuated influenza vaccine (LAIV) do not provide cross-protection against drift and potential pandemic strains. We hypothesized that supplementing LAIV with a highly conserved protective antigenic target M2 ectodomain (M2e) would confer broad cross-protection by inducing humoral and cellular immune responses to conserved antigenic targets (M2e, NP), overcoming strain-specific protection of current vaccines. Intranasal vaccination with LAIV (A/NL/09/H1N1) supplemented with tandem repeat M2e containing virus-like particles (M2e5x VLPs) induced M2- and H1N1 virus-specific antibodies. Upon heterosubtypic challenge with pandemic potential rgA/H5N1 virus, we found that M2e5x VLP supplemented LAIV immune mice showed significantly improved heterosubtypic protection by preventing weight loss and lowering lung viral titers compared to the LAIV alone group inducing poor heterosubtypic immunity. Further mechanistic studies on heterosubtypic immunity suggest that recall T cell responses to M2e and nucleoprotein as well as systemic and mucosal M2e antibodies were found to be major heterosubtypic immune correlates. Therefore, this study demonstrates a novel vaccination strategy to confer universal protection by supplementing current strain-specific vaccines with a highly conserved cross protective antigenic target M2e5x VLPs.
Therapeutic platform for immunotherapy of Chagas disease (VAC9P.1106)
Authors: Konduri Vanaja , Liang Dan (Vivian) , Halpert Matthew , Levitt Jonathan , Hotez Peter , Jones Kathryn , Decker William 
Published: January 2023

Abstract
Chagas Disease, caused by infection with the protozoan parasite Trypanosoma cruzi, is a leading cause of heart disease and sudden cardiac death in Latin America, disproportionately affecting people living in resource-poor areas. Approved treatment is limited to two drugs, benznidazole and nifurtimox, which have poor efficacy in the chronic stage of disease as well as severe side effects. Thus, there is an urgent need for alternative therapies to treat Chagas Disease. Pre-clinical models have demonstrated that antigen specific CD8+IFNγ+ responses are essential for reducing parasite burdens, increasing survival, and decreasing cardiac pathology in both the acute and chronic phases of Chagas. In the present study, we developed an adenovirus-based gene transfer cassette that was used therapeutically to prevent or delay the cardiac complications that result from infection with T. cruzi. The therapeutic cassette, containing T. cruzi Tc24 and TSA1 antigens and a dominant-negative isoform of the SHP-1 inhibitory phosphatase, mediated development of antigen-specific CD8+IFNγ+ cells when ex vivo-transduced dendritic cells were administered to Balb/c mice. Additionally, the vaccine platform reduced parasite burdens, increased survival, and reduced cardiac pathology in a Balb/c model of live T. cruzi infection. The data suggest that immunotherapeutic approaches might be successful in addressing the unmet medical need for improved Chagas treatments.
Azithromycin inhibits IL-1 secretion and non-canonical inflammasome activation (VAC9P.1110)
Authors: Gualdoni Guido , Lingscheid Tilman , Schmetterer Klaus , Hennig Annika , Steinberger Peter , Zlabinger Gerhard 
Published: January 2023

Abstract
Deregulation of inflammasome activation was recently identified to be involved in the pathogenesis of various inflammatory diseases. Although macrolide antibiotics have well-established immunomodulatory properties probably involved in their clinical effects, studies on their impact on this pathway are missing. Comparing the influence of macrolides on cytokine induction in human monocytes, we found that azithromycin, but not clarithromycin or roxithromycin, specifically inhibited IL-1β secretion upon LPS stimulation whereas no effects were observed after flagellin stimulation. Blockage of azithromycin’s transmembrane transport abolished the cytokine-modulatory effect, indicating intracellular azithromycin accumulation to be causative for diverging effects. Consistent with IL-1β inhibition restricted to LPS stimulation, we found impaired induction of the intracellular LPS-sensing caspase-4 whereas NF-κB signalling was unaffected. Substantiating our in vitro data, azithromycin specifically dampened IL-1β levels and enhanced survival in a murine endotoxin sepsis model. Overall, we provide first evidence of a differential impact of macrolides on the inflammasome/IL-1β axis, which might be of relevance in inflammasome driven diseases like COPD or asthma.
Fc receptor γ chain plays a critical role in lung viral clearance and reducing vaccine-enhanced disease in mice immunized with respiratory syncytial virus vaccines (VAC9P.1063)
Authors: Hwang Hye suk , Lee Young Tae , Kim Ki-Hye 
Published: January 2023

Abstract
Vaccine-enhanced respiratory disease (ERD) is well-known for respiratory syncytial virus (RSV) after vaccination with formalin inactivated RSV (FI-RSV). Induction of Th2 type immune responses and deposition of immune complexes have been suggested to be possible mechanisms. We hypothesized that Fc receptor γ chain (FcRγ) would play a role in conferring protection against RSV after vaccination and that priming with Th1 inducing vaccine would prevent ERD. Virus-like particles presenting RSV fusion protein (F VLP) were found to induce Th1 type immune responses. Wild type (WT) mice and FcRγ KO mice were immunized with FI-RSV or primed with F VLP followed by FI-RSV boost (F VLP/FI-RSV). RSV specific serum IgG and isotype antibody levels were similar in both WT and FcRγ KO mice after RSV vaccination. However, F VLP/FI-RSV FcRγ KO mice showed lower levels of RSV neutralizing antibody titers and IFN-γ secreting CD8+ T cells, higher lung viral titers and more body weight loss compared to F VLP/FI-RSV WT mice. Heterologous F VLP/FI-RSV immunization induced Th1 type humoral and cellular immune responses, no overt sign of ERD in the bronchoalveolar lavage airways in both WT and FcRγ KO mice compared to homologous FI-RSV immunization. The results in this study support a critical role of FcRγ in lung viral clearance and diminishing ERD. More importantly, a strategy of priming with Th1 inducing vaccine might be a promising approach to induce protection against RSV and to prevent ERD.
Preclinical evaluation of a trivalent vaccine for treating/preventing candidal disease (VAC9P.1066)
Authors: Schmidt Clint , DeMontigny Elizabeth , Lizakowski Mary , Zacher Erica , Hoeg Jesse , Cochrane Terrence , Hennessey John 
Published: January 2023

Abstract
Multiple-antigen vaccines are an increasing focus of biotherapeutic development targeting infectious disease. Three recombinant antigens, based on Candida albicans proteins, have independently induced protection in animal models of candidal infections; agglutinin-like sequence 3 (rAls3), hyphally regulated protein 1 (rHyr1), and secreted aspartyl proteinase 2 (rSap2). However, a trivalent vaccine with all 3 antigens has not been previously evaluated. Here we present murine immune data for monovalent and multivalent vaccine formulations, with and without aluminum hydroxide (AlOH). IgG titers are induced against each antigen in the trivalent vaccine, and all are higher with AlOH. Interference is seen with anti-rAls3 IgG titers when equal amounts of all 3 antigens are used. This interference is abrogated by decreasing rHyr1 and rSap2 amounts relative to rAls3. Dose-dependent IL-17A splenocyte responses were detected against rAls3 and rHyr1, but not rSap2, in monovalent formulations. With trivalent formulations, Als3- and Hyr1-specific IL-17A production increased additively in magnitude and numbers of mice responding and at lower doses compared to monovalent formulations. These data support that robust immune responses can be induced by all 3 antigens in a trivalent vaccine, which may improve efficacy for some or all disease indications by targeting multiple facets of the candidal pathogenesis pathway, including adhesion, invasion and dissemination.
TCR gene expression in lymphoid and mucosal tissues of Xenopus laevis (VET1P.1122)
Authors: Geibel Gretchen , Kongprachaya Yuthana , Appasamy Pierette 
Published: January 2023

Abstract
γδ T cells are a small subset of T cells that are considered to function as a bridge between innate and adaptive immunity. Because these cells are relatively few in number in most laboratory animals, except during the fetal period and in mucosal tissues, it is difficult to clearly delineate the functions of γδ T cells. In order to begin to characterize the γδ T cell populations in Xenopus laevis, previous work from this lab established a semi-quantitative RT-PCR method of analysis of TCR γ and TCR β gene expression in X. laevis thymus and spleen. We have extended these studies to evaluation of expression of TCR γ, β, α and δ transcripts in the thymus and spleen as well as the skin, intestine and lungs of young adult X. laevis. Based on evaluation of TCR transcript levels by RT-PCR, γδ T cells are likely to be abundant in the mucosal tissues evaluated, and the levels of TCR γ are higher in the X. laevis thymus and spleen than expected for adult humans or mice. We suggest that X. laevis is a γδ T cell “high” organism, perhaps related to the position of frogs as relatively early in the evolution of the adaptive immune response. Evaluation of expression of other T cell-related genes including the RAG genes and other genes involved in T cell development and function is ongoing. These results point to the use of X. laevis as a uniquely powerful laboratory animal model for studies of the evolution, development and function of γδ T cells.
Investigating distinct immunization routes in zebrafish on mucosal immunity (VET2P.1133)
Authors: Criscitiello Michael , Young Matthew , Troy Savannah , Cory Hope , McCauley Naomi , Al-Kahtani Sibba , Morgan Thomas , Scholtz Sheridan , Li Ellen , Ali Hasanain , Bains Sarab , Briles Brenna , Chang Jonathan , Cruz-Vespa Diego , Diaz Navarro Jean , Eberle Brady , Gonzalez Allison , Marvin Sophia , Nelson Mary , Roubion Courtney , Watts Jiang , Chen Pat , Basler Julie , Minckler Zachary , Wu James 
Published: January 2023

Abstract
Model organisms such as Danio rerio are used to explore physiological reactions to stress and disease. Immune response is measured by quantifying lymphocyte presence and the production of mRNA encoding different immunoglobulin proteins. Since the method through which D. rerio is exposed to an antigen affects its immune response and the degree to which immunity is developed, we will compare the immune response after visceral or mucosal introduction of dinitrophenol hapten conjugated to Keyhole Limpet hemocyanin (DNP-KLH) in D. rerio, for which there is no published data. We will either combine DNP-KLH with Freund’s incomplete adjuvant and inject the mixture into the fish (100 uL at 1 mg/mL) or combine DNP-KLH with cholera toxin and introduce the mixture to their water (35 mg/L). Tissue samples will be collected from the immunized fish and two control groups of fish at weekly points throughout the experiment and will be analyzed for lymphocyte versus granulocyte ratios and expression of IgM, IgZ-1, and IgZ-2 levels via qPCR. We expect an increase in lymphocyte percentages for both immunized groups compared to the control groups, with a higher lymphocyte ratio in the mucosal group than the visceral group. Furthermore, we expect more mucosal IgZ isotypes in the mucosal group and higher levels of the IgM, IgZ-1, and IgZ-2 antibodies in the visceral group than in the mucosal group. For future experiments, we intend to test broader data sets, including exposure to multiple antigens.
Ion efflux and influenza infection trigger NLRP3 inflammasome signaling in human dendritic cell (VIR5P.1148)
Authors: Fernandez Melissa , Miller Elizabeth , Krammer Florian , Greenbaum Benjamin , Bhardwaj Nina 
Published: January 2023

Abstract
The NLRP3 inflammasome is an essential intracellular mediator of anti-viral immunity. In murine dendritic cells (DCs) this complex responds to a wide array of signals, including ion efflux and influenza A virus (IAV) infection, to activate caspase-1 mediated proteolysis of interleukin (IL)-1β and IL-18 into biologically active cytokines. However, the presence and function of the NLRP3 inflammasome in human DCs in response to various triggers, including viral infection, has not been clearly defined. Here, we delineate the contribution of the NLRP3 inflammasome to the secretion of IL-1β, IL-18, and IL-1α by human DCs. Activation of the NLRP3 inflammasome in human DCs by various synthetic activators resulted in the secretion of bioactive IL-1β, IL-18, and IL-1α, and induction of pyroptotic cell death. Cellular IL-1β release depended on potassium efflux, and the activity of proteins NLRP3 and caspase-1. Similarly, IAV infection of DCs resulted in priming and activation of the NLRP3 inflammasome and secretion of IL-1β and IL-18 in an M2 and NRLP3 dependent manner. The magnitude of priming by IAV varied amongst different strains and inversely corresponded to type I IFN production. To our knowledge, this is the first report describing the existence and function of the NLRP3 inflammasome in human DCs, and the ability of IAV to prime and activate this pathway in human DCs with important implications for anti-viral immunity and pathogenesis.
LIGHT programs activated CD8 T cells to survive and become memory cells in response to a viral pathogen (VIR5P.1150)
Authors: Desai Pritesh , Tahiliani Vikas , Abboud Georges , Hutchinson Tarun , Ware Carl , Croft Michael , Salek-Ardakani Shahram 
Published: January 2023

Abstract
CD8 memory T cells play a critical role in protection against repeated exposure to respiratory viruses. Memory T cell development in the lung is still not fully understood in terms of the nature and source of the molecular signals that establish and maintain the memory state. LIGHT is a tumor necrosis factor family member expressed by immune cells such as activated T cells, dendritic cells, monocytes and granulocytes. It is known to interact with two receptors HVEM & LTβR, leading to differential outcomes. However, the role of LIGHT expressed specifically by CD8 T cells during respiratory viral infection is not known. Using adoptive transfer of LIGHT-deficient CD8 TCR-transgenic T cells responding to Ag in the context of respiratory vaccinia virus (VACV) infection, we found that LIGHT signaling in CD8 T cells had a small impact on the level of primary expansion, with at most 30-40% fewer CD8 T cells accumulating in the lungs at the peak of response to VACV infection. In striking contrast, very few effector CD8 T cells that lacked LIGHT survived the contraction phase to differentiate into long-lived memory cells in the lungs. Furthermore, we found that HVEM, but not LTβR, functions as a trans-activating binding partner for LIGHT expressed on CD8 T cells. These results underscore the importance of LIGHT in the memory CD8 T cell development and suggest that targeting LIGHT-HVEM interaction will help in designing CD8 T cell based vaccines against respiratory viruses.
Lymphotoxin β receptor signaling promotes influenza-induced lung damage (VIR5P.1151)
Authors: Spencer Cody , Koroleva Ekaterina , Muraoka Wayne , Blackman Marcia , Tumanov Alexei 
Published: January 2023

Abstract
Lymphotoxin β receptor (LTβR) has been shown to play a critical role in protection against mucosal bacterial pathogens in the gut. Surprisingly, we found that genetic or biochemical inhibition of LTβR signaling confers protection against highly pathogenic H1N1 influenza virus infection. Consistent with amelioration of disease severity, LTβR-deficient mice displayed reduced expression of pro-inflammatory cytokines, reduced inflammatory dendritic cells, and drastically reduced numbers of antigen specific CD8+ T cells in the lung. Further, LTβR-deficient mice showed attenuated viral loads during the peak of viral replication. Previous studies by our group have highlighted a potential role of LT signaling in the regulation of mucin expression in the gut. Further, increased mucin secretion in the lung is known to correlate with attenuation of influenza virus replication. Immunohistochemical and gene expression analysis revealed a marked increase in goblet cell hyperplasia and MUC5AC secretion in the lungs of influenza infected LTβR-deficient mice relative to wild type mice. Additionally, treatment of infected wild type mice with an agonistic anti-LTβR antibody resulted in almost complete inhibition of MUC5AC mRNA expression in the lung. Taken together, our data suggest that LT signaling negatively regulates mucin production in response to respiratory viral infection leading to increased viral burden, lung pathology, and death.