bluuebunny/crossref_metadata_2025_split huggingface dataset:
FcRn overexpression in transgenic animals results in robust immune response (P1069)
Authors: Kacskovics Imre , Schneider Zita , Vegh Attila , Ilias Attila , Baranyi Maria , Balogh Peter , Cervenak Judit , Kurucz Istvan 
Published: January 2023

Abstract
Our previous studies have shown that overexpression of the neonatal Fc receptor (FcRn) in transgenic animals (mice and rabbits) leads to (1) improved IgG protection; (2) larger numbers of antigen specific B cells; (3) generation of antibodies against weakly immunogenic antigens; (4) generation of antibodies against highly conserved antigens (breaking immune tolerance); (5) increased humoral diversity and (6) higher levels of circulating antibodies. FcRn transgenesis thus confers a number of practical benefits, including faster antibody production, higher antibody yields and improved generation of hybridomas for monoclonal antibody production. To gain additional insights into the mechanisms underlying this increase in humoral immune response, we further characterized our FcRn transgenic animals. Our analyses showed strong expression of the FcRn transgene in professional antigen presenting cells and we also found that overexpression of the FcRn significantly improves antigen presentation in case of antigen-IgG immune complexes. We have recently analyzed several key factors in the maturation of the immune response, including the structure of the spleen, number and structure of germinal centers, the level of T cell activation and the affinity of the generated antibodies. These results provide further insights of those mechanisms that are involved in the augmented humoral immune response in transgenic animals that overexpress FcRn.
Detection of phosphorylated signaling proteins using phospho-specific monoclonal antibodies (P1115)
Authors: Miletic Ana , Funatake Castle , Scott Bradley , Alvarez Miguel , Chen Helen , Li Shourong , Schrantz Nicolas , Just Peggy 
Published: January 2023

Abstract
Triggering of extracellular surface receptors results in the initiation of numerous intracellular signaling cascades that are propagated via phosphorylation of specific tyrosine, serine, and/or threonine residues on signaling proteins. Until recently, analysis of intracellular phospho-signaling relied solely upon western blotting. The development of phospho-specific antibodies for use in flow cytometry has become a powerful tool for the detection of signaling events and has numerous advantages over western blotting; it is faster, requires fewer cells, and can be performed in a heterogeneous population of cells in conjunction with surface marker staining to allow further phenotyping of each population present in the sample. To this end, we have developed monoclonal antibodies specific for phosphorylation of various signaling molecules such as ERK1/2 (T202/Y204), STAT1 (Y701), STAT4 (Y693), and STAT5 (Y695). Here, we show the detection of phosphorylation of different intracellular signaling by flow cytometry across species in a treatment- and cell type-specific manner. One issue complicating phospho-specific flow cytometry is the inability to detect certain surface markers because of either epitope or fluorochrome destruction using the current, standard staining protocol. We show options that allow multi-parametric staining with different phospho-specific antibodies.
A West Nile Virus NS4B P38G mutant strain induces adaptive immunity via TLR7-MyD88-dependent and independent signaling pathways. (P4317)
Authors: Xie Guorui , Welte Thomas , Wang Jia , Wang Ye , Whiteman Melissa , Wicker Jason , Saxena Vandana , Cong Yingzi , Barrett Alan , Wang Tian 
Published: January 2023

Abstract
West Nile virus (WNV)- induced neurological disease has become a public health concern. No vaccines have been approved for human use. We have previously shown that an attenuated WNV, the nonstructural (NS) 4B-P38G mutant, induced stronger innate cytokine and T cell responses than wild-type WNV. Mice immunized with this mutant were all protected from subsequent lethal wild-type WNV infection. Here, we found both Toll-like receptor (TLR7) and MyD88 expression are required for host protection from NS4B P38G mutant infection. Infected MyD88-/- and TLR7-/- mice had lower IL-12 expression, IgM production, and reduced effector T cell functions. MyD88-/- and TLR7-/- dendritic cells (DC)s exhibited impaired antigen-presenting functions compared to wild-type DCs. At the late stage of infection or during secondary challenge with wild-type WNV, MyD88-/- and IL-1R-/- mice displayed reduced memory T cell responses compared to wild-type mice; whereas TLR7-/- mice had normal T cell functions. Moreover, immunization of MyD88-/- and TLR7-/- mice with NS4B P38G mutant strain offer partial or full protection respectively from subsequent lethal wild-type WNV infection. Thus, TLR7-MyD88 signaling is required for initial T cell priming; whereas the TLR7-independent MyD88 signaling is involved in memory T cell development during NS4B P38G mutant infection. Results can be utilized as a paradigm to aid in the rational development of efficacious live attenuated flavivirus vaccines.
Prime-boost vaccination with a live attenuated influenza vaccine elicits T cell-mediated cross-strain protective immunity (P4287)
Authors: Zeng Mingtao , Li Junwei , Arévalo Maria , Chen Yanping 
Published: January 2023

Abstract
Antigen drift and shift of influenza viruses requires frequent reformulation of influenza vaccines. This stresses the need for a universal influenza vaccine. In this study, mice were vaccinated with the trivalent live attenuated (LAIV, nasal spray FluMist) and inactivated (TIV, flu shot FluZone) influenza vaccines by 1-3 dose prime-boost vaccination regimens. While vaccination with both LAIV and TIV elicited strong humoral responses, only LAIV induced significant protection against challenge with the non-vaccine strain H1N1 PR8 influenza virus. Furthermore, protective immunity against PR8 was dose-dependent. Of note, IL-2 and IFN-γ cytokine secretion in the lung alveolar fluid were significantly elevated in mice immunized with LAIV. Moreover, T cell depletion of LAIV immunized mice compromised protection, indicating that T cell-mediated immunities were required. In contrast, passive transfer of sera from mice immunized with LAIV into naïve mice failed to protect against PR8 challenge. To verify these results, we performed virus neutralization assays in vitro and confirmed that LAIV did not induce cross-strain neutralizing antibodies against PR8 virus. Finally, we showed that 3 doses of LAIV also provided protection against challenge with additional two heterologous viruses FM/47 (H1N1) and HK/68 (H3N2). These results support the potential use of the LAIV as a universal influenza vaccine under prime-boost vaccination regimen.
Cholera toxin breakdowns oral tolerance via activation of canonical NF-κB (P3208)
Authors: Im Suhn-Young , Kim Kyoung-Jin , Kim Han-A , Lee Hern-Ku , Seo Kook Heon 
Published: January 2023

Abstract
The mechanisms of mucosal immunogenicity and adjuvanticity of bacterial exotoxins remain to be established. In this study, we investigated the ability of cholera toxin (CT) to induce the transcription factor NF-κB. Feeding of CT abrogated OVA-induced oral tolerance, as evaluated by OVA-specific serum Ab responses, footpad swelling reaction (FSR), and CD4+ T cell proliferation. CT feeding resulted in clear NF-κB activation in both mesenteric lymph node (MLN) and Peyer’s patch (PP) cells. Supershift analysis demonstrated that mainly one heterodimer type, p50-p65, i.e., canonical NF-κB, occurred in response to CT. In addition to NF-κB activation, CT induced the expression of mRNA in NF-κB-dependent proinflammatory cytokines, such as IL-1β, IL-6, and TNF-α in vivo as well as in vitro. CT no longer showed abrogation of oral tolerance in mice pretreated with p50 small interfering RNAs (siRNAs), but not in control siRNA. ADP-ribosylation inhibitors inhibited CT-induced NF-κB activation in MLN and PP intestinal lymphoid cells. These data suggest that CT induces canonical NF-κB activation in intestinal lymphoid cells, which plays a key role in mucosal immunogenicity and adjuvanticity.
TAM receptors and their role in the regulation of lung innate immunity (P3236)
Authors: Fujimori Toshifumi , Snelgrove Robert , Hussell Tracy 
Published: January 2023

Abstract
Background: The role of TAM receptors are well characterised in autoimmunity and cancer biology. Their role in infectious diseases is now being elucidated. The goal of this study is to characterise TAM receptor expression profiles of various innate immune components at homeostasis as well as in various infection models. The hypothesis of this study is that up-regulation of TAM receptors, AXL and MerTK are responsible for the de-sensitisation of airway macrophages to bacteria following influenza infection. Furthermore, I hypothesise that TAM receptors may be involved in limiting innate immune activation at homoeostasis. Methods: Using a murine model, we characterised TAM receptor expression on various innate populations in the lung following viral (influenza A) and bacterial (Streptcoccus pneumoniae) infections. We also investigated the role of AXL in influenza/bacteria single and co-infection using AXL-/- mice. Results: AXL and MerTK are differentially expressed by innate populations in the lung depending on their location. Monocytes and neutrophils which infiltrate into the alveolar space during influenza infection significantly up-regulate TAM receptor expressions. Lack of AXL significantly potentiates innate immunity by upregulating the release of inflammatory cytokines in vitro. Conclusion: Up-regulation of AXL following a viral infection may explain the exacerbation of secondary bacterial complications as its upregulation limits the innate inflammatory responses.
CXCR5-CXCL-13-pathway in protective immunity to tuberculosis (P3190)
Authors: Khader Shabaana , Slight Samantha , Gopal Radha 
Published: January 2023

Abstract
One third of the world’s population is infected with Mycobacterium tuberculosis (Mtb). Although most infected people remain asymptomatic, they have a 10% lifetime risk of developing active tuberculosis (TB). Thus, the current challenge is to identify immune parameters that distinguish individuals with latent TB from those with active TB. Using human and experimental models of Mtb infection, we show that organized ectopic lymphoid structures containing CXCR5+ T cells are found in Mtb-infected lungs. In addition, we show that in experimental Mtb infection models, the presence of CXCR5+ T cells inside ectopic lymphoid structures are associated with immune control. Furthermore, in a mouse model of Mtb infection, we show that activated CD4+ CXCR5+ T cells accumulate in Mtb-infected lungs, and produce proinflammatory cytokines. Absence of CXCR5 results in increased susceptibility to TB due to defective T cell localization within the lung parenchyma. We show that CXCR5 expression on T cells mediates correct T cell localization within TB granulomas, efficient macrophage activation, promoting protection against Mtb infection and facilitating lymphoid follicle formation. We also show that inducing T helper 17 (Th17) memory responses using mucosal immunization strategies improves vaccine-induced immunity against TB highlighting the potential use of IL-17-CXCR5-CXCL-13 axis in future TB vaccine design and therapy.
Non-covalent association of protein and capsular polysaccharide on bacteria-sized latex beads as a model for polysaccharide-specific humoral immunity to intact Gram-positive extracellular bacteria (P3083)
Authors: Colino Jesus , Duke Leah , Snapper Clifford 
Published: January 2023

Abstract
Intact Streptococcus pneumoniae, expressing type 14 capsular polysaccharide (PPS14) and type 3 Streptococcus agalactiae containing a PPS14 core capsule identical to PPS14, exhibit non-covalent associations of PPS14 and bacterial proteins, in contrast to covalent conjugates. Secondary immunization with conjugate and S. agalactiae, although not S. pneumoniae, results in a boosted response. However, in contrast to conjugate, PPS14-specific IgG responses to bacteria lack affinity maturation and use the 44.1-idiotype. To better understand the mechanism underlying this dichotomy we developed a minimal model of intact bacteria utilizing 1μm latex particles to which PPS14 and pneumococcal surface protein A (PspA) were stably attached, but not directly linked, in contrast to PPS14-PspA conjugate. PPS14+[PspA] beads, similar to conjugate, induced in mice boosted PPS14-specific IgG secondary responses, dependent on T cells and ICOS-dependent costimulation, and in which priming could be achieved with PspA alone. In contrast to conjugate, but similar to bacteria, the primary PPS14-specific IgG response to PPS14+[PspA] beads peaked rapidly, with the secondary response highly enriched in 44.1-idiotype and lacking affinity maturation. These results demonstrate that non-covalent association in a particle, of polysaccharide and protein, recapitulates essential immunologic characteristics of intact bacteria that are distinct from soluble covalent conjugates of these respective antigens.
IL-1α and IL-1β play non-redundant roles in limiting in vivo Aspergillus fumigatus pulmonary growth (P3092)
Authors: Obar Joshua , Lehmann Margret , Caffrey Alayna , Watschke Christopher , Shepardson Kelly , Barker Bridget , Cramer Robert 
Published: January 2023

Abstract
Aspergillus fumigatus is a mold that causes severe pulmonary infections in humans, such as invasive pulmonary Aspergillosis (IPA). Currently, our knowledge of how A. fumigatus growth is controlled in the respiratory tract is limited. Phagocytic alveolar macrophages constitute the first line of defense against inhaled A. fumigatus conidia; subsequently, neutrophils and macrophages are sequentially recruited to the respiratory tract to control fungal growth and germination . But how neutrophils and macrophages are recruited to the respiratory tract after A. fumiguatus infection remains ill defined. During sterile inflammation early neutrophil recruitment is dependent on IL-1α, while late macrophage recruitment is dependent on IL-1β. A. fumigatus infection induced the expression of both IL-1α and IL-1β. However, IL-1R1- and ASC-deficient mice displayed differential susceptibility to IPA. IL-1R1-deficient mice were highly susceptible to A. fumigatus infection as measured by increases in fungal growth, lung tissue damage, and fungal dissemination. In contrast, ASC-deficient mice were only mildly susceptible to A. fumigatus infection. IL-1R1- and ASC-deficient mice displayed a severe defect in neutrophil recruitment during the first 48h and a defect in macrophage recruitment after 48h, respectively. Taken together, our data reveal important non-redundant roles for the IL-1α and IL-1β in controlling A. fumigatus infection in the lung.
Highly immunogenic vaccine for prevention and therapy of malignant mesothelioma (P4437)
Authors: De Groot Anne , Bertino Pietro , Hoffman Peter , Carbone Michele , Siccardi Antonio , Terry Frances , Cousens Leslie , Moise Leonard , Martin William 
Published: January 2023

Abstract
Malignant mesothelioma (MM) is a deadly cancer with increasing incidence and no effective treatment options. We have previously constructed recombinant Fowlpox virus (FP) vectors encoding full-length survivin protein (FP-surv), which induced complete tumor regression in 40% of mice bearing subcutaneous MM. Macro- and microscopic evaluation of the tumor tissue showed that the tumors in vaccinated mice had large necrotic cores resulting from enhanced survivin-specific cytotoxic T lymphocyte (CTL). We aim to refine our novel FP vaccine by focusing on the most immunodominant survivin epitopes, thus enhancing its efficacy. We have identified additional high quality antigenic targets specifically upregulated in MM that will increase the breadth of T cell responses. We have used the EpiMatrix system to identify CD4+ and CD8+ T cell epitopes for synthesis and testing. Following T cell assays, highly immunogenic peptides will be selected that specifically stimulate CTLs, without activating T regulatory cells (Treg) that interfere with anti-cancer immune responses. We will include the selected peptides in the next-generation vaccine, FP-surv.2, which we expect to be more effective for inducing an immune response against MM cells than FP carrying the entire survivin gene (FP-surv). In addition, we anticipate that this response may include a higher number of long-lived memory T cells that can be exploited to prevent MM development in high-risk predisposed subjects.
The clinical implication of HAV viral load and HAV proteins-specific IFN-gamma production in patients with acute hepatitis A (P6120)
Authors: Lee Hyun Woong , Chang Dong-Yeop , Shin Eui-Cheol 
Published: January 2023

Abstract
The relationship between immune responses to hepatitis A virus (HAV) and the clinical manifestation of HAV infection have been obscured. We investigated the clinical manifestation of acute hepatitis A (AHA) and the relationship between HAV-specific, IFN-gamma-producing T cell response and clinical outcomes in patients with AHA. We analyzed 770 patients diagnosed with AHA for their clinical manifestation. Among them, 128 patients with a prothrombin time (PT) of less than 40% of normal were classified as “severe acute hepatitis (s-AH)”. Others were defined as having “mild acute hepatitis (m-AH)”. In 110 patients, direct ex vivo IFN-gamma ELISpot assays were performed with peripheral blood mononuclear cells using HAV proteins. Mean values of serum HAV RNA at admission were 4.79 log10 copies/ml in s-AH, and 3.30 log10 copies/ml in m-AH (P = 0.003). Viral load showed a significant correlation with ALT (r = 0.417, P = 0.0032) and PT levels (r = -0.453, P = 0.0012). In addition, the mean frequencies of IFN-gamma-producing T cells against VP1-2A (aa 669-782), and 2C(aa 1121-1234) proteins were significantly lower in patients with s-AH, compared to patients with m-AH (3.3 vs. 17.7, P &lt; 0.001; 2.1 vs. 13.3, P &lt; 0.001). In view of viral factors, the load of HAV is closely correlated with liver damage and disease severity in patients with AHA. As a host factor, HAV-specific, IFN-gamma-producing T cells might induce effective viral eradication and result in mild hepatitis
Gut bacteria modulates angiogenesis and corneal immunopathology after herpes simplex virus infection (P6136)
Authors: Richardson Raphael , Bhela Siddheshvar 
Published: January 2023

Abstract
The ocular disease herpetic stromal keratitis results in blindness due to an immunopathological attack mainly orchestrated by T cells and neutrophils. Recently, it has become apparent that lesion severity at local sites may be influenced by the balance of microbes in distal locations such as the gut, respiratory tract and even the skin. Reasons are still being assembled but appear to involve an influence on both innate and adaptive immunity. Since HSK is an uncommon outcome of ocular HSV infections in the infected individual, we sought to determine if the nature of enteric flora would influence the expression of HSK. A murine model of HSK was used to evaluate the concept. Mice were ocularly infected with HSV and separated into two groups. The controls received only sucrolose while the experimental group received sucrolose and a combination of antibiotics (Ampicillin, Gentamicin, Metronidazole, Neomycin, and Gentamicin) 3 weeks before infection to deplete gut flora and treatment continued until termination of the experiment at day 15. Significant decreases in corneal angiogenesis and HSK was evident in the antibiotic treated recipients. Analysis of inflammation cell numbers and molecules involved in angiogenesis also showed significant differences between treated and control groups. Further mechanistic studies to explain for the differences observed will I be reported and discussed.
Role of peripheral immune response in regulating chemokine and pro-inflammatory cytokine profiles during acute viral encephalitis (P6147)
Authors: Steel Christina , Brevis Kimberly , Tavakoli Susan , Kim Woong-Ki , Sanford Larry , Ciavarra Richard 
Published: January 2023

Abstract
Vesicular stomatitis virus (VSV) is a neurotropic virus that rapidly invades the central nervous system (CNS) following a single intranasal application and induces acute encephalitis essential for survival. Invasion of the CNS by inflammatory cells is thought to reflect virus-induced production of chemokines. To assess this possibility and to broadly characterize this response during the initial and acute phases of infection, brain chemokine and pro-inflammatory cytokine RNA profiles were determined in naïve and VSV-infected mice using qPCR arrays and flow cytometry. RNA transcript levels for several CC, CXC chemokines and cytokines were already elevated one day post infection with several ligand/receptor pairs co-induced which precedes peripheral leukocyte recruitment to the CNS. During acute VSV encephalitis (days 6-8), RNA levels became more robust with expression of additional chemokines and cytokines and a mixed leukocyte (previously reported). Chemokine responses associated with VSV encephalitis were blunted in mice acutely depleted of either peripheral dendritic cells or T cells. Several CC receptors for induced chemokines were highly expressed on microglia isolated from encephalitic brains. Infiltrating neutrophils and T cells failed to express these same receptors. These results suggest that different cytokines and/or chemokines may be responsible for recruitment of peripheral leukocyte subsets into the CNS, while others may drive the observed microgliosis.
Inflammasome regulates host immune responses to enteric pathogen (P4237)
Authors: Liu Zhiping , Kanneganti Thirumala-devi 
Published: January 2023

Abstract
Inflammasome is emerging as a central regulator of inflammatory immune responses. Nlrp3 inflammasome has been shown to be critical for intestinal homeostasis and inflammation using chemical dextran sodium sulfate -induced colitis model. However, the roles of inflammasome in other more natural models of colitis such as colitis induced by enteric bacteria (e.g., Citrobacter rodentium) are not well known. Our study identified that mice lacking the inflammasome components Nlrp3, Nlrc4 and Caspase-1 were highly susceptible to C. rodentium-induced colitis, demonstrated by more weight loss, higher bacterial burden and more severe histological lesions, in comparison to wild-type (WT) mice. Inflammasome-deficient mice displayed much higher infiltration of neutrophil and macrophage into spleen and colon. In addition, in comparison to WT mice, Caspase 1-deficient mice had higher percentage of active T cell that secret more IFN-γ, which might significantly contribute to the elevated inflammation in intestinal mucosa. We also identified that inflammasome-mediated production of IL-1β and IL-18 are critical for host defense against C. rodentium infection. Dissect the mechanisms how these inflammatory cytokines regulate host immune responses to enteric pathogens will be of great interest to the understanding of inflammatory bowel disease pathogenesis.