bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enhanced protection for influenza vaccines using a novel rOv-ASP-1 adjuvant aqueous formulation (VAC2P.933)
Authors: Jiang Jiu , Fisher Erin , Hensley Scott , Lustigman Sara , Murasko Donna , Shen Hao 
Published: January 2023

Abstract
Adjuvants have been used to enhance the efficacy of a variety of vaccines; however, no adjuvant is included in current influenza vaccines used in the United States. In this study, we found that a novel adjuvant, rOv-ASP-1, co-administrated with inactivated influenza vaccine using an aqueous formulation, substantially improved the influenza-specific antibody response and protection against lethal infection in a mouse model. rOv-ASP-1 enhanced the magnitude of the specific antibody response after immunization with low doses of influenza vaccine, allowing antigen-sparring by 10-fold. The rOv-ASP-1 formulated vaccine induced a more rapid response and a stronger Th1-associated antibody response compared to vaccine alone and to the vaccine formulated with the adjuvant alum. Importantly, rOv-ASP-1 significantly enhanced cross-reactive antibody responses and protection when challenged with a heterologous influenza virus strain. These results demonstrate that rOv-ASP-1 is an effective adjuvant that: 1) accelerates and enhances the specific antibody response induced by influenza vaccine; 2) allows for antigen sparing; and 3) augments a Th1-biased and cross-reactive antibody response that confers heterologous protection.
Circulating CXCR5+PD-1+ response predicts influenza vaccine antibody responses in young adults but not older adults (VAC2P.936)
Authors: Canaday David , Herati Ramin , Reuter Morgan , Dolfi Douglas , Mansfield Kathleen , Kurupati Raj , Ertl Hildegund , Schmader Kenneth , Kannan Senthil , Betts Michael , Wherry John 
Published: January 2023

Abstract
Although influenza vaccination is recommend for all adults annually, the incidence of vaccine failure, defined as weak or absent increase in neutralizing antibody titers, is increased in the elderly compared to young adults. The T follicular helper subset of CD4+ T cells provide B cell help in germinal centers and are necessary for class-switched antibody responses. Previous studies suggested a role for circulating T follicular helper cells (cTfh) following influenza vaccination in adults, but cTfh have not been studied in elderly adults where weak vaccine responses are often observed. Here, we studied cTfh expressing CXCR5 and Programmed Death 1 (PD-1). cTfh from elderly adults were present at reduced frequency, had decreased in vitro B cell help ability, and greater expression of activation marker inducible costimulator (ICOS) compared to young adults. At seven days after inactivated influenza vaccination, cTfh correlated with influenza vaccine-specific IgM and IgG responses in young adults but not in elderly adults. In sum, we have identified aging-related changes in cTfh which manifested as reduced influenza vaccine responses. Future rational vaccine design efforts should incorporate Tfh measurement as an immune correlate of protection, particularly in the setting of aging.
Vaccination with DHA-derived specialized proresolving mediators increases the antibody-mediated immune response against influenza virus (VAC2P.925)
Authors: Ramon Sesquile , Baker Steven , Sahler Julie , Feldsott Eric , Martínez-Sobrido Luis , Topham David , Serhan Charles , Phipps Richard 
Published: January 2023

Abstract
Influenza viruses remain a critical global health concern. More efficacious vaccines are needed to protect against influenza virus, yet few adjuvants are approved for routine use. Specialized proresolving mediators (SPMs) are powerful endogenous bioactive lipid regulators of inflammation, with great clinical translational properties. We previously identified endogenous levels of the SPM resolvin D1, protectin D1 and 17-hydroxydocosahexaenoic acid (17-HDHA) in the mouse spleen. Here, we investigated the ability of 17-HDHA to enhance the adaptive immune response using a pre-clinical influenza vaccination mouse model. Our findings revealed that mice immunized with influenza H1N1-derived HA protein plus 17-HDHA increased antigen specific antibody titers. 17-HDHA increased the number of plasma cells and HA-specific antibody-secreting cells in the bone marrow, but it did not affect the frequency of GC B cells or Tfh cells in the draining lymph node. Importantly, the 17-HDHA-mediated increased antibody production was more protective against live pH1N1 influenza virus infection in mice. This is the first report on the immuno-stimulatory effects of omega-3-derived SPMs on the humoral immune response. These findings illustrate a previously unknown biological relation between proresolution signals and the adaptive immune system. In addition, 17-HDHA as well as other immuno-stimulatory SPM have the potential to be used as novel endogenous adjuvants.
VacSIM™ - a new delivery method increases influenza vaccine efficacy (VAC2P.928)
Authors: Samli E. Farah , Shollenberger Lisa , Harn Donald 
Published: January 2023

Abstract
Influenza is a global health issue causing considerable economic burden. Seasonal epidemics of this acute viral infection spread rapidly with unpredictable morbidity. Regular vaccination remains the most effective prevention and as such, there is tremendous need for new technologies focused on improving immunogenicity and efficacy of seasonal flu vaccines. VacSIM™, which stands for Vaccine Self-assembling Immune Matrix, represents a new method of vaccine delivery incorporating an inert biopolymer with unique biophysical properties. It can be stored at 4-22°C, as a viscous-liquid and is flexible enough to incorporate various antigens, adjuvants or organisms for vaccine delivery. Post-injection VacSIM™ undergoes a phase shift to a semi-solid gel, forming a temporary depot of concentrated vaccine components. This gel-matrix depot may slow clearance of the vaccine, simultaneously enabling time-released delivery of the vaccine. We believe the latter increases activation of antigen presenting cells. Mice vaccinated with whole inactivated influenza (A/Puerto Rico/08/34) or with recombinant nucleoprotein via VacSIM™, had an increased humoral response, greater protection from lethal influenza challenge and improved viral clearance from lungs. Studies with other candidate vaccine antigens suggest VacSIM™ is capable of enhancing the immunogenicity and efficacy of a wide variety of vaccines.
B7-H1 blockade improves efficacy of regulatory T cell depletion as cancer immunotherapy by reducing Treg regeneration, possibly through tumor B7-H1 effects (VAC3P.950)
Authors: Curiel Tyler , Sareddy Gangadhara , Hurez Vincent , Qin Kunhua , Pandeswara Sri Lakshmi , Vadlamudi Ratna , Murthy Kruthi 
Published: January 2023

Abstract
Immune co-signaling blockade is effective cancer immunotherapy but is not curative alone. Treg depletion has shown potential human anti-cancer efficacy, but is also not curative alone. As B7-H1 immune co-signaling facilitates Treg generation, we hypothesized that adding αB7-H1 to Treg depletion would improve treatment by hindering Treg regeneration. We show here that αB7-H1 plus denileukin diftitox (DD) to deplete Tregs is a synergistic combo that effects complete and durable (&gt;7 mos) tumor regressions in bulky B16 melanoma, whereas neither agent alone does. αB7-H1 efficacy alone or with DD is lost in IFN-γ KO or T cell KO mice, and is mediated in part by tumor-specific CD8+ (but not CD4+) T cells. αB7-H1 is highly effective in B7-H1 KO mice, demonstrating a major role for tumor B7-H1 in treatment efficacy. Reducing tumor B7-H1-mediated Treg generation appears to be a mechanism as predicted. The αB7-H1/DD combo was also effective in ID8 ovarian cancer showing broad application. We engineered B7-H1hi/lo B16 tumors for further immune studies. Surprisingly, B7-H1hi B16 grew faster than B7-H1lo in RAG KO, suggesting B7-H1 as an adaptive immunity-independent tumor growth/survival factor. IFNγ upregulated tumor B7-H1, but αB7-H1 did not prevent IFN-γ-mediated tumor inhibition in vitro. B7-H1hi/lo tumors in B7-H1 KO are being used to define tumor vs host effects of αB7-H1 ± Treg depletion. As αB7-H1 and DD have been tested clinically, this combo is rapidly clinically translated.
Rescue of vaccine T cell memory following sublethal gamma irradiation (VAC3P.952)
Authors: Berkson Julia , McFarland Hugh , Rosenberg Amy 
Published: January 2023

Abstract
Subsequent to sublethal γ radiation exposure, survivors reacquire immune competence, but lose vaccine immunity. Our goal is to rescue CD8+ T cell-mediated memory responses following γ radiation exposure. In a mouse model of Listeria monocytogenes (LM) infection, we found that rapid revaccination (rescue) of immunized mice following irradiation preserves CD8+ T cell mediated immunity. Rescued mice are able to survive a wild type LM challenge infection when the rescue vaccine strain, LM-deltaActA, is given 0-3 days following radiation exposure. A signature of rescue was revealed in gene profiling studies in which a group of cytokines and chemokines were uniquely up regulated in the spleens of LM-rescued animals as compared to mice irradiated without rescue. These included factors important in resistance to LM, such as proinflammatory factors CCL2, TNFα, and IL-6 and bacterocidal factors CXCL9, 10, and 11 as well as anti-apoptotic factors including IL-15 and IER3, which may have a role in rescue of memory CTL. RAW264.7, a macrophage cell line, produced a similar cytokine profile when irradiated and infected with LM, suggesting a probable myeloid source for the majority of these factors. Monocyte activation by the combination of irradiation and LM infection may thus create conditions of rescue by producing cytokines and chemokines that enhance clearance of LM and rescue CD8+ CTL mediating vaccine immunity.
Potency and stability of LAP+FOXP3+ Tregs in control GVHD development (VAC3P.954)
Authors: Tran Dat , Wang Huihui , Pham Anha , Cooper Laurence 
Published: January 2023

Abstract
Intro: We have developed previously a method to isolated bona fide Tregs from expansion cultures based on their selective surface expression of latency-associated peptide (LAP). In this study, we investigated their in vivo potency and stability using a humanized GVHD murine model. Methods: CD25+ cells isolated from PBMC via AutoMACS were stimulated with OKT3 loaded K562 aAPC expressing CD64 and CD86 in media +IL2 and rapamycin. 36h post-restimulation at wk2, the Tregs were re-isolated with antiLAP via AutoMACS. The LAP+ Tregs and LAP- nonTregs were expanded for 2wks. In vitro suppression, phenotype and TSDR demethylation were performed after 4wk expansion. CD25-depleted human PBMC were injected into NSG mice to induce xGVHD and compared with coinjection of LAP+ Tregs or LAP- nonTregs. Results: LAP+ repurification results in &gt;85% LAP+FOXP3+ Tregs, leaving behind FOXP3- and FOXP3+ nonTregs within the LAP- population. After a total of 4wk expansion, LAP+ Tregs were &gt;1 billion, &gt;85% FOXP3, highly suppressive and anergic in vitro, and &gt;95% demethylated TSDR (p&lt;0.05). In GVHD model, PBMC alone resulted in a median survival of 29 days. The LAP+ Tregs increased median survival to 43 days (p&lt;0.01), while the LAP- non-Tregs had a median survival of 28.5 days. Conclusions: LAP+ Tregs are highly purified, stable, and potent based on these preclinical studies. LAP+ Tregs are an ideal immunotherapeutic population for clinical trials to prevent or treat GVHD and other autoimmune diseases.
Endogenous NK cell HMGB1 is central to HDIL-2 induced systemic autophagy (VAC3P.946)
Authors: Liang Xiaoyan , LI Guanqiao , Vernon Philip , Buchser William , Loughran Patricia , Basse Per , Zeh Herbert , Lotze Michael 
Published: January 2023

Abstract
Administration of high dose IL-2 (HDIL-2) has durable antitumor effects in 8-10% of patients with metastatic melanoma and RCC but is associated with substantial toxicity. We explored the underlying mechanisms of this toxicity. Administration of HDIL-2 (600KIU/mouse) significantly enhances serum level of the inflammatory cytokines IFN-γ, IL-6, IL-18 and HMGB1. These increases are associated with increased LC3-II in multiple organs in B6 mice. This is consistent with the notion that HDIL-2 induces systemic autophagy. These responses are lethal in RAG1-/- mice. IL-2 treatment with chloroquine (CQ, 50mg/kg/d) enhances survival in RAG1-/- mice from 20% in the IL-2 alone cohort to 82% in mice receiving CQ+IL-2 (p=0.01). Surprisingly, NK cell depletion provides 100% survival in HDIL-2 treated RAG1-/- mouse (p&lt;0.01). Although no HDIL-2 toxicity was observed in NSG mice, this could be reversed with the adoptive transfer of NK cells. Serum levels of cytokines were significant enhanced (all p&lt;0.05): HMGB1 (41.9±13.5 to 256.6±26.8 ng/ml), RAGE (2.4±0.5 to 15.8±0.5 ng/ml), IFN- γ (0±18 to 420±0 pg/ml), IL-18(205±19.7 to 2318±430 pg/ml). Interestingly, treatment with α-HMGB1 Ab conferred protection. Combinations of IL-2 (200K IU/mouse) and IL-18 (25ug/kg/d X5) induced lethal systemic autophagy in WT mice. This was significantly abrogated in mice with HMGB1 genominically ablated in NKp46+ cells. Thus, NK cell plays a critical role in HDIL-2 toxicity and endogenous HMGB1 may be a key factor.
Induction and protective effect of innate interferons during human astrovirus infection (VIR1P.1003)
Authors: Cline Troy , Huerta Teddy , Freiden Pamela , Fugate Nicole , Schultz-Cherry Stacey 
Published: January 2023

Abstract
Members of the relatively new Astroviridae family are non-enveloped, single-stranded RNA viruses that cause diarrheal disease in a wide range species. Human astroviruses, of which there are eight canonical strains, are a leading cause of gastroenteritis in children &lt;2 years old, the elderly, and immunocompromised patients. Because of the lack of a small animal model of astroviral gastroenteritis, little is known about the host response to infection. The role of type I and type III interferons (IFN), IFN-α/β and IFN-λ respectively, in mediating innate antiviral immune responses through a STAT1-dependent mechanism have been well established. Using human astrovirus type 1 (HAstV-1), the most common serotype isolated from human cases of gastroenteritis, we investigated the innate IFN response to astrovirus in cultured colonic epithelial cells (Caco2). HAstV-1 induced modest levels of IFN-β in Caco2 cells by 24 hours post-infection. Pre-treatment of cells with IFN-β protected Caco2 cells from HAstV-1 infection in a dose-dependent manner. Further, viral titers were significantly elevated in the presence of a neutralizing antibody to IFN-β. These data are in contrast to IFN-λ, which is induced by HAstV-1 to levels approximately 10-fold greater than those of IFN-β. However, IFN-λ pretreatment was not protective. Consistent with this, IFN-λ treatment did not induce STAT1 phosphorylation, suggesting that Caco2 cells are not responsive to IFN-λ.
Identifying the DHHC palmitoyl acyltransferase responsible for activating the antiviral protein IFITM3 (VIR1P.1008)
Authors: McMichael Temet , Hach Jocelyn , Chesarino Nicholas , Yount Jacob 
Published: January 2023

Abstract
Influenza virus poses a significant health threat to the global population. The intrinsic immune factor, interferon inducible transmembrane protein 3 (IFITM3) has been shown to inhibit the infection efficiency of all tested influenza virus subtypes, and understanding its mechanism of action and post-translational regulation may provide new strategies for combating infections. Palmitoylation represents a post-translational modification that positively affects IFITM3 membrane affinity and is necessary for its complete anti-influenza virus activity. Palmitoylation is generally carried out by palmitoyl acyltransferases (PATs) that have conserved DHHC amino acid motifs. Currently, it is not known which PATs are responsible for the palmitoylation of IFITM3. Our preliminary screening, using a PAT overexpression assay and chemical reporters to measure IFITM3 palmitoylation, has implicated four candidate PATs. Quantification of IFITM3 antiviral activity against influenza virus upon PAT overexpression has implicated a single PAT able to increase IFITM3 activity. Confocal imaging of this PAT demonstrated a unique pattern of localization among the other candidates suggesting that the site of IFITM3 palmitoylation is a critical factor in controlling its activity. Identifying the PAT responsible for palmitoylating IFITM3 should promote our understanding of the post-translational regulation, cellular trafficking, and activity of this important antiviral effector.
Inhibition of Langerhans cell maturation by human papillomavirus type 16: a novel role for the annexin A2 heterotetramer in immune suppression (VIR1P.961)
Authors: Kast Wijbe , Woodham Andrew , Raff Adam , Raff Laura , Yan Lisa , Skeate Joseph , Wong Michael , Lin Yvonne , Da Silva Diane 
Published: January 2023

Abstract
Human papillomaviruses (HPV) are associated with several cancers. During its life cycle HPV16 infects the epithelial basal cells through a receptor we recently identified, the annexin A2 heterotetramer (A2t). During infection, HPV16 also interacts with Langerhans cells (LC), the antigen presenting cells of the epithelium, and induces immune suppression mediated by the HPV16 L2 minor capsid protein. Despite the importance of these virus-immune cell interactions, the specific mechanisms of HPV16 entry into LC and HPV16-induced immune suppression remain undefined. An N-terminal peptide of HPV16 L2 has been shown to specifically interact with A2t. Here, we show that incubation of human LC with this peptide blocks binding of HPV16. Inhibiting this interaction with an A2t ligand or by siRNA down-regulation of A2t, significantly decreases HPV16 internalization into LC in an L2-dependent manner. A2t is associated with inhibition of LC maturation as demonstrated through attenuated secretion of Th1-associated cytokines and decreased surface expression of MHC II on LC exposed to A2t. Conversely, small molecule inhibition of A2t prevents HPV16-induced inhibition of LC immune function as indicated by increased secretion of inflammatory cytokines and surface expression of MHC II and CD86 in HPV16 treated LC pre-exposed to A2t inhibitors. These results demonstrate that HPV16 suppresses LC maturation through an interaction with A2t, revealing a novel immune suppressive role for this protein.
Licensed NK-mediated response to murine cytomegalovirus specifically upregulates dendritic cell co-stimulatory ligands required in T cell priming (VIR1P.963)
Authors: Teoh Jeffrey , Stadnisky Michael , Bullock Timothy , Brown Michael 
Published: January 2023

Abstract
Ly49G2+ (G2+) NK cells mediate murine (M)CMV resistance in mice with the self-MHC Dk ligand expressed. A prior study showed that G2+ NK-mediated MCMV resistance resulted in increased numbers of conventional dendritic cells (cDC) and virus-specific CD8 T cells after infection, independent of CD4 T cells. However, the molecular basis underlying licensed NK-mediated enhancement of T-cell immunity is unknown. Here we examined the role of co-stimulatory ligands expressed in splenic cDC after MCMV infection. An increase in the intracellular expression of both CD86 and CD70 was observed by 72-hr, first in the CD8α+ DC and then in the CD11b+ DC. To test functionality, we used CD86 and CD70 mAbs to block co-stimulation before infection. CD86-block had no effect on CD8 T cells. In contrast, CD70-block impaired both the frequency and the numbers of MCMV-specific CD8 T cells without perturbing cDC numbers, cDC maturation, or G2+ NK-mediated viral control. We next examined the importance of CD70-CD27 signaling in CD27-/- mice with the self-ligand Dk expressed. We observed similar deficits in Ag-specific CD8 T cells, despite having intact G2+ NK-mediated MCMV control. Moreover, CD8 T cells in both CD70-blocked and CD27-/- infected mice responded to MCMV peptide re-stimulation with less degranulation and cytokine production. We infer that a licensed G2+ NK-mediated response to MCMV upregulates co-stimulatory molecule expression in cDC that is needed to efficiently prime CD8 T cells.
Targeting Robo4 pathway reduces corneal angiogenesis induced by herpes simplex virus. (VIR1P.967)
Authors: Gimenez Fernanda , Rouse Bary 
Published: January 2023

Abstract
The cornea is a complex tissue that must preserve its transparency to maintain optimal vision. Eye infections such as with herpes simplex virus (HSV) can result in a chronic immuno-inflammatory syndrome and pathological neovascularization that impairs vision. Recent reports demonstrate that angiogenic sprouting shows morphological similarities to axon growth. Moreover there are several key molecules that upon binding to receptors regulate the direction of both capillary and axon guidance. In this study we show that Roundabout 4 (ROBO4) receptor plays a significant role reducing angiogenesis. Using ROBO4 -/- mice we demonstrate that these mice are hyper-susceptible to HSV induced corneal angiogenesis compared to wild type mice. Moreover, providing additional soluble ROBO4 protein by subconjunctival administration significantly diminished the extent of corneal angiogenesis in wild type animals. Finally, by administration of soluble ROBO4 protein to ROBO-/- HSV infected mice it was possible to rescue the wild type phenotype. In conclusion, the administration of soluble ROBO4 protein promotes the activation of the ROBO4 pathway. This could represent a valuable therapeutic tool to control corneal angiogenesis related to HSV induced stromal keratitis.
Defective viral genomes arising in vivo promote the onset of lung antiviral immunity (VIR2P.1023)
Authors: Lopez Carolina , Tapia Karla , Kim Won-keun 
Published: January 2023

Abstract
In infections with viruses well adapted to the host, virus-encoded proteins that delay the cellular response allow the virus to replicate to high titers prior to host intervention. The mechanisms overcoming viral evasion and leading to the production of the primary antiviral cytokine IFNbeta are not well established. We show that the appearance of defective forms of the viral genome may provide the answer. Defective viral genomes, which were until recently considered an epiphenomenon of in vitro virus replication, accumulate in infected cells at a faster rate than standard viral genomes and potently trigger the sustained activation of the transcription factors IRF3 and NF-kB and the production type I IFNs through a mechanism independent of IFN signaling. Remarkably, our work demonstrates that defective viral genomes are generated naturally during respiratory infections in vivo even in mice lacking the type I IFN receptor, and their appearance coincides with the production of cytokines during infections with Sendai virus or influenza virus. Notably, the hallmark antiviral cytokine IFNβ is only expressed in lung epithelial cells containing defective viral genomes, while cells within the lung that contain standard viral genomes alone do not express this cytokine. Together, our data indicate that defective viral genomes generated naturally during viral replication are a primary source of danger signals for the initiation of the host immune response to infection in vivo.