bluuebunny/crossref_metadata_2025_split huggingface dataset:
Essential role of E3 ubiquitin ligase activity of GRAIL in T cell functions (P1111)
Authors: Nurieva Roza , Wang Junmei , Alekseev Andrei 
Published: January 2023

Abstract
T cells are the master regulators of adaptive immune responses, and many autoimmune diseases arise due to a breakdown of self-tolerance in T cells. Understanding of the molecular mechanisms underlying T cell tolerance will lead to development of pharmacological approaches either to promote the tolerance state in terms of autoimmunity or to break tolerance in cancer. E3 ubiquitin ligases have been placed among the essential molecules involved in the regulation of T cell functions and T cell tolerance. We as well as other groups have reported that T cells activated in the absence of both CD28 and ICOS costimulation developed into tolerant T cells, associated with markedly upregulated expression of the E3 ubiquitin ligase GRAIL. In order to understand the physiological function of GRAIL, we generated mice deficient in Grail by replacing region that encompassing most of the RING domain and responsible for E3 ubiquitin activity. Remarkably, genetic inactivation of E3 ubiquitin ligase function of GRAIL led to T cell hyper-responsiveness to TCR/CD3z signaling and their independency to costimulation for activation. As a result, GRAIL-deficient mice were more predisposing to autoimmune diseases. On the other hand, modulation of GRAIL function helped to boost T cell immune responses to cancer, and, therefore, mediate tumor rejection. Thus, modulation of the E3 ligase activity of GRAIL might be an important approach to control T cell functions in autoimmunity or cancer.
CD43 activates moonlight functions of pyruvate kinase (P1113)
Authors: Bravo-Adame Maria Elena , Vera-Estrella Rosario , Barkla Bronwyn , Cruz-Muñoz Mario , Rosenstein Yvonne 
Published: January 2023

Abstract
CD43 is the most abundant costimulatory molecule on the T cell surface; it transduces activation signals through its cytoplasmic domain, modulating the outcome of T cell responses. CD43 KO mice revealed a positive role for CD43 in T cell migration and in the regulation of cell number and homeostasis. The aim of this study was to identify new signaling pathways modulated by CD43. Protein expression profiles of normal human CD4+ T cells stimulated through TCR and CD43 identified changes in 189 proteins. Bioinformatic analysis revealed that glucose metabolism was significantly represented. We evaluated pyruvate kinase (PK) and lactate dehydrogenase (LDH) activities, as well as glucose uptake. TCR+CD43 stimulated cells showed a 1.5 fold increase in PK activity, but only a moderate increase in LDH activity and glucose uptake. Interestingly, tyrosine phosphorylation of PK triggers its ability to phosphorylate Stat3. Phosphorylation of Y105 of PK and of Y705 of Stat3 was induced in response to TCR+CD43 and TCR+CD28. Since phosphorylation of Y705 of Stat3 by PK induces MEK5 transcription, we evaluated the MEK5/ERK5 pathway activation by measuring the levels of pCREB at S133, an ERK5 target. CREB pS133 was increased in TCR+CD43 and TCR+CD28 stimulated T cells. Altogether, our results show that CD43 regulates monosaccharide metabolism, as well as moonlight functions of metabolic enzymes.
The direct impact of key cytokines on memory effectors from older adult human CD8+ T cells to live influenza virus (P4297)
Authors: Zhou Xin , Swain Susan , McElhaney Janet 
Published: January 2023

Abstract
An age-related decline in cytolytic activity has been described in CD8+ T cells and we have shown that the poor CD8+ effector T cell response to influenza A/H3N2 challenge is due to a decline in the proportion and function of these cytolytic T lymphocytes (CTL). Here we report that there is a significant decline in the levels of many cytokines and chemokines including IFN-γ, IL-2, IL-10 and IL-17A, but increased IL-4 and IL-6, following influenza-stimulation of PBMCs from vaccinated older compared to young adults. We find that addition of exogenous IL-2 acts to maintain naïve CD8+ T cells survival and strongly enhances the proliferation of existing memory effector CD8+ T cells in response to live influenza virus challenge. In addition exogenous IL-6 plays a similar but lesser role in the proliferative response when compared to IL-2. Our data show that only exogenous IL-4 enhances the proliferative capacity of the CD8+ T cells and particularly the promotion of naïve CD8+ T cell differentiation. However, IL-4 treatment has no effect on the accumulation of CD8+ effector T cells in response to influenza challenge. These results suggest mechanisms that might be exploited to develop influenza vaccines with improved efficacy in older adults.
Attenuated dendritic cell function in BALB/c mice following HPAI vaccination results from failed C-type lectin receptor signaling (P4309)
Authors: Vaughan Sarah , Stout-Delgado Heather , Pollock Zemmie , Plourde Jennifer , Pyles John , Harrod Kevin 
Published: January 2023

Abstract
Highly pathogenic avian influenza (HPAI) has an approximate 60% fatality rate in humans. In the United States, a vaccine for HPAI has been manufactured and stockpiled using FDA approved methods for seasonal vaccines. Compared to seasonal vaccines, the HPAI vaccine is less immunogenic in humans despite similar production methods. We developed a mouse model of influenza vaccination to examine differences in the host immune response to these vaccines. Mice that received the seasonal vaccine produced a robust neutralizing antibody response whereas no neutralizing antibodies were detected following HPAI vaccination. Dendritic cells (DCs) cultured with HPAI vaccine had decreased expression of activation markers CD40 and CD86, and produced significantly lower levels of cytokines IL-6, IL-12, and TNF-α compared to those treated with seasonal vaccine. C-type lectin receptors have been implicated as a class of pattern recognition receptors in innate immunity and immunization. Pretreatment with mannan diminished cytokine induction by DCs in a dose dependent manner following seasonal but not HPAI vaccine treatment, suggesting a role for C-type lectin receptors in DC activation by influenza vaccines. These findings implicate a potential mechanism for attenuated DC function following HPAI vaccination and may explain the lack of immunogenicity of the currently approved HPAI vaccine.
Recombinant Ebolavirus antigens from insect cells are potent immunogens inducing cellular and humoral immunity in rodents and non-human primates and provide protection against virus challenge (P4325)
Authors: Lieberman Michael , Lehrer Axel , Humphreys Tom , Johns Lisa , Wong Teri-Ann , Olinger Gene , Dye John , Carrion Ricardo , Patterson Jean , Marzi Andrea , Feldmann Heinz 
Published: January 2023

Abstract
Soluble recombinant Filovirus surface glycoproteins (GP) and matrix proteins (VP24 and VP40) were generated in the Drosophila S2 cell expression system and purified by immunoaffinity chromatography. The immunogenicity of individual recombinant Zaire ebolavirus (ZEBOV) subunits and admixtures with or without adjuvants was evaluated in mice, guinea pigs and macaques. Strong antigen-specific IgG responses were observed, includng Ebola virus neutralization responses. In mice and macaques subunit proteins were shown to elicit cell mediated immunity, as significant B- and T-cell stimulation was observed in immune lymphocytes after antigen re-stimulation. Analysis of secreted cytokines in batch-cultured, antigen-stimulated splenocytes or PBMC’s demonstrated Th1 and Th2 type responses. Vaccine candidates were tested in mice and guinea pigs for direct protection against challenge with species-adapted ZEBOV. All vaccine formulations containing ZEBOV GP were protective in mice and serum transfer from such animals into naïve mice demonstrated that humoral immunity alone can be fully protective. Furthermore, the transfer of immune splenocytes into naïve mice showed that recombinant GP and VP24 subunits elicit functional T cell responses that lead to protection against live virus challenge. In guinea pigs, lead vaccine formulations consistently produced high antibody responses and demonstrated 100% protective efficacy in the ZEBOV challenge model.
Heterologous prime-boost vaccination with DNA vaccine and recombinant subunit containing four serotypes of dengue virus envelope protein domain III elicits neutralizing antibodies and specific T-cell responses (P4327)
Authors: Pan Chien-Hsiung , Hu Hui-mei , Hsiao Yu-Ju , Wu Sze-Hsien 
Published: January 2023

Abstract
Dengue is still an important public health problem and no dengue vaccine is available now. In this study, we characterized prime-boost vaccine regimens using heterologous and homologous dengue DNA vaccine and recombinant subunit vaccine consisted of envelope (E) protein domain III (ED III) from four serotypes of dengue virus (DENV). A broader and balanced IFN-gamma and IL-4 responses was observed in DNA-prime and subunit-boost immunized mice, compared to a narrower but strong IFN-gamma production after homologous DNA vaccination and an IL-4 only response in homologous subunit vaccine immunized mice. In addition, a significant ED III specific antibody response was detected in all immunized mice except vector control. The highest IgG titer was appeared in homologous subunit vaccine immunized mice, followed by heterologous prime-boost regimen and homologous DNA vaccine. However, that heterologous prime-boost immunized mice generated a highest neutralizing antibody than the others, suggested a better protection provided by this DNA-prime and subunit vaccine-boost regimen.
SeVRSV, a PIV and RSV vaccine, proves efficacious in a cotton rat maternal antibody model (P4279)
Authors: Hurwitz Julia , Portner Allen , Russell Charles , Surman Sherri , Jones Bart , Sealy Robert , Rudraraju Rajeev , DeVincenzo John 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) and the human parainfluenza viruses (PIV) are causes of significant morbidity and mortality among infants each year. We are now advancing SeVRSV, a Sendai virus based Jennerian vaccine for human PIV-1 and a recombinant vaccine for RSV. SeVRSV expresses RSV F and induces immunity that protects against RSV in small animals and non-human primates. Immune responses persist systemically and in the respiratory tract, which ensures that pathogens can be blocked at their point-of-entry. One concern surrounding pediatric vaccines is that maternal antibodies may obstruct vaccine efficacy. We therefore designed a cotton rat maternal antibody model to test SeVRSV. First, to define the levels of maternal antibodies typical of the vaccine's intended target population, we titered antibodies from approximately 100 human infants. Then, by passive transfer techniques, we matched these titers in animals. Intranasal vaccination in test animals was followed by the measurement of de novo PIV-specific and RSV-specific antibodies, and cotton rat challenge with RSV to measure protection. In models using polyclonal or monoclonal antibody products with specificities for PIV and/or RSV, the SeVRSV vaccine protected against RSV challenge. Data suggest that SeVRSV will not be inhibited by maternal antibodies in the clinical setting, and encourage rapid development of the vaccine for protection of human infants from PIV and RSV.
CpG vaccination induces cytolytic T cells to the lung and heterosubtypic protection against lethal IAV infection (P4289)
Authors: Vogel Alexander , Condon Shirley , Brown Deborah 
Published: January 2023

Abstract
With the emergence of pandemic influenza A virus (IAV) strains such as H1N1 and H5N1, the development of a universal vaccine has become more necessary than ever. In contrast to the highly mutable outer coat proteins, internal proteins of IAV are more conserved among different strains and are therefore a target for developing vaccines that induce strong T cell responses. Using a Toll-like receptor 9 agonist with unmethylated CpG motifs, we have shown that intranasal immunization with CpG and inactivated x31 (H3N2) IAV protects mice following a heterosubtypic challenge with a lethal dose of PR8 virus (H1N1). Vaccination with CpG and inactivated IAV cause a stronger inflammatory response than IAV infection alone, with significant levels of antiviral cytokines such as MIP-1β, CXCL 10, and IL-6 produced in the lungs. Furthermore, CpG immunization induces a substantial influx of CD11b+ cells into the draining lymph nodes as well as a population of CD11c+ cells that express high levels of the costimulatory molecule CD86, providing favorable conditions for T cell activation. At 6 days post immunization, effector CD4 and CD8 T cells expressing Granzyme B and IFN-γ are observed in the lungs of vaccinated mice. These experiments suggest that TLR agonists can act as potent immune adjuvants to generate robust T cell immunity capable of combating IAV and other viral infections where successful vaccines have not yet been developed.
Network analysis of lung tissue repair processes in response to influenza-induced damage identifies tissue remodeling factor TIMP-1 in recovery (P3180)
Authors: Sanders Catherine , Sharma Shalini , Vogel Peter , Doherty Peter , Thomas Paul 
Published: January 2023

Abstract
Mild and severe influenza infections clearly differ in disease resolution, likely due to variations in the host’s repair response to lung tissue damage. Over 60 protein factors involved in lung tissue repair at multiple stages of a mild and a severe influenza infection (early, mid-stage, viral clearance, and after viral clearance) were quantified and organized into an interaction network. Factors characteristic of all phases of tissue repair—coagulation, inflammation, migration and proliferation, and remodeling—were up-regulated in mouse whole lung tissue after both types of infections. Differences in intensity and timing of these factors, particularly in the later three stages of wound repair, were observed. Network analysis allowed for identification of a key node in the repair pathway important in tissue remodeling, TIMP-1. TIMP-1-/- mice were more susceptible to PR/8 infection independently of viral titer or inflammatory immune responses. However, respiratory analyses showed exacerbated responses to both infections, suggestive of dysregulated lung function during attempted resolution of disease. We demonstrate the utility of systems biology for understanding complex, multi-layered, integrated biological processes such as wound healing, and in identification of previously overlooked factors involved in such processes. We also demonstrate the contribution of TIMP-1, a protein not involved in viral control, coagulation, or inflammation, in recovery from influenza infection.
Establishment of non-circulating, tissue-resident anti-viral memory T cells in specific niches of the lung (P3198)
Authors: Turner Damian , Bickham Kara , Farber Donna 
Published: January 2023

Abstract
Virus-specific memory T cells residing in mucosal sites play an important role in conferring immune protection against recurrent infections. Here we describe the specific retention of memory T cell populations within distinct lung niches following influenza virus (IAV) infection. We used intravenous infusion of fluorescent antibody to differentiate between T cells in the lung interstitium (protected) and those accessible to circulation (labeled). While naïve mice had a negligible frequency of CD4 T cells in the protected niche, after IAV infection 50-70% of total lung CD4 T cells segregated into the protected niche with 30-40% stably maintained within this niche as memory T cells after viral clearance. IAV-specific memory CD4 and CD8 T cells were highly enriched in the protected population compared to circulating T cells. Protected T cells did not recirculate, unlike labeled memory T cells which continuously circulate through lymphoid tissues and are significantly reduced in the lung when lymphoid egress is inhibited. Confocal microscopy showed that effector and memory CD4 T cells in the protected niche were clustered around airways and bronchovascular bundles, while the labeled CD4 T cells were dispersed randomly within the capillaries of the alveoli walls. Our results demonstrate that, following IAV infection, virus-specific memory T cells are retained in spatially distinct lung niches which may be important for protective immunity to respiratory pathogens.
The role of CXCR6 in lung airway memory CD8 T cell recruitment and maintenance (P3202)
Authors: McMaster Sean , Wilson Jarad , Wang Hong , Kohlmeier Jacob 
Published: January 2023

Abstract
Tissue-resident memory T (TRM) cells are uniquely positioned to provide an initial line of defense against invading microorganisms; airway-resident memory CD8 T (TARM) cells have been shown to confer protective immunity against a number of respiratory pathogens, including influenza virus and M. tuberculosis. Unlike TRM at other anatomical sites, TARM are not maintained via cytokine-driven homeostatic proliferation in the airways but must be continuously recruited from the circulation. We show that there exists a defect in TARM cell recruitment in mice infected with influenza virus via either intramuscular or intraperitoneal route as compared to the intranasal route. However, the mechanism(s) that enable intranasally-primed memory T cells to preferentially migrate to the airways have not been identified. Based on our preliminary data, we hypothesize that the candidate chemokine receptor, CXCR6, is responsible for steady-state trafficking of influenza-specific TARM cells to the airways from the general circulation. Our results show that CD8 T cell recruitment to the lung airways is significantly restricted in CXCR6 KO mice as compared to WT, while CD8 T cell numbers in the spleen remain constant. This study could help advance the field of cell-mediated vaccines to promote pathogen-specific T cell recruitment to the lung airways, enhancing protective immunity to respiratory pathogens.
Resistance to invasive pneumococcal infection in mice with acute allergic lung inflammation (P3228)
Authors: Sanfilippo Alan , Metzger Dennis 
Published: January 2023

Abstract
Streptococcus pneumoniae is the most common cause of bacterial pneumonia which results in 175,000 hospitalizations as well as 40,000 deaths in the United States annually. Recent work has identified a link between asthma and invasive pneumococcal disease (IPD) prompting the Advisory Committee on Immunization Practices to recommend that all adults with asthma receive vaccination. To study the underlying mechanism responsible for an increased risk of IPD for individuals with asthma, we used an acute model of ovalbumin (OVA)-induced allergic lung inflammation (ALI) and hypothesized that mice with acute ALI would be more susceptible than naive mice to a pulmonary S. pneumoniae infection. Surprisingly, mice with acute ALI were more resistant to infection than naïve controls and this increased resistance was found to be greatest 10 days following the final OVA challenge. While inflammatory cell infiltration and cytokine levels in the bronchoalveolar lavage (BAL) fluid had returned to levels similar to naïve controls by day 10, this time point also correlated with the peak of IgA expression in the BAL fluid, suggesting a critical role for non-specific IgA in protection against IPD. Indeed, protection against IPD was lost in IgA KO mice with ALI as well as naïve IgA KO mice.
Thymic stromal lymphopoietin directly influences the respiratory CD8 T cell response to influenza virus infection. (P3230)
Authors: Shane Hillary , Klonowski Kimberly 
Published: January 2023

Abstract
Mucosal surfaces are tightly regulated immune microenvironments with unique anatomical structures, profiles of cells and cytokines. These aspects, coupled with infection derived inflammatory signals, can differentially modulate downstream immunological responses. Thymic stromal lymphopoietin (TSLP) is a cytokine expressed in mucosal surfaces where its role in CD4 T cell immunity has been well studied. However, whether TSLP directly affects anti-viral mucosal CD8 T cell responses is less understood. Here, we show that influenza induces TSLP production in a lung epithelial cell line and infected mice. To test the direct role of TSLP on mucosal CD8 T cell responses we used a competitive adoptive transfer system where equal numbers of OVA-specific OT-I and TSLP receptor deficient OT-I cells are co-transferred into recipient mice infected with influenza expressing OVA. We found that TSLPR-/- OT-I CD8 T cells proliferate less early post infection (pi) and have decreased accumulation in the respiratory tract both at days 8 and 50pi. Interestingly, the lack of TSLP signaling imparted a higher level of CD62L expression on TSLPR-/- OT-I memory cells. Together these data suggest that TSLP signaling may directly drive the expansion and development of CD8 effector phenotypes while concomitantly limiting long term responses. Thus, TSLP participates in the regulation of immune responses in the respiratory tract and modulation of TSLP levels may promote long term immunity at mucosal sites.
The induction of humoral immune responses to the orally administered Dukoral vaccine occurs independently of TLR signaling and DC maturation (P3188)
Authors: Kumar Ashok , Majithia Jay , Azizi Ali , Sirskyj Danylo 
Published: January 2023

Abstract
We investigated the involvement of TLR signaling in the induction of humoral and cell-mediated immune responses (CMIR) following oral and intramuscular (IM) immunization with Dukoral (killed whole-cell V. cholerae and cholera toxin B subunit, CTB) or its components by using TLR-2-, TLR-4-, MyD88- and Trif-deficient mice. We show that wild type and all groups of TLR-deficient mice generated similar levels of V. cholerae- and CTB-specific IgG1and IgG2c serum antibodies, fecal IgA antibodies and agglutinating antibodies, following oral immunization with Dukoral or CTB alone. However, IM immunization with Dukoral or CTB alone required MyD88 signaling for the induction of CTB-specific IgG1and IgG2c serum antibodies. In addition, our results show that CD4+ T-cell and CD19+ B-cell proliferation, maturation of DCs in response to vaccine components, as measured by CD80, CD86, CD40, and MHCII expression, and cytokine secretion including IFN-g occurred in a MyD88-dependent manner in response to V. cholerae or CTB stimulation. In contrast, in response to CTB stimulation, Trif negatively regulated both CD4+ T-cell and CD19+ B-cell proliferation. Overall, our results suggest that despite the impaired ability of MyD88-/- DCs to mature, humoral responses, following oral immunization with Dukoral, occurred independently of TLR signaling, while CMIR were TLR-dependent. However, TLR-independence is lost in the generation of humoral responses when the vaccine is administered parenterally.