bluuebunny/crossref_metadata_2025_split huggingface dataset:
Resveratrol protects experimental Staphylococcus aureus enterotoxin B (SEB)-induced acute liver injury (ALI) by regulating microRNAs associated with myeloid-derived suppressor cells
Authors: Singh Narendra Prasad , Kadhim Sabah , Singh Udai P. , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Resveratrol (RES) possesses anti-inflammatory, anticancer, and antioxidant properties. Because of anti-inflammatory property of RES, we investigated whether RES can attenuate Staphylococcal enterotoxin B (SEB)-induced acute liver injury (ALI) in mice. SEB exposure causes food poisoning as well as toxic shock syndrome leading to mortality. We observed that SEB caused ALI in female mice and there was an increase in enzyme aspartate transaminase (AST) levels, induction of inflammatory cytokines, and massive infiltration of immune cells into the liver. RES (50 mg/kg body weight) treatment attenuated SEB-induced ALI and significantly decreased AST levels, inflammatory cytokines and chemokines, and cellular infiltration in the liver. Interestingly, RES treatment led to an increase in myeloid derived suppressor cells (MDSC) in SEB-immunized mice. Upon examination of microRNAs (miRs) profile of liver infiltrating mononuclear cells (MNCs), there was marked change in microRNA expression profile in mice exposed to SEB and treated with RES. Specifically, there was a decrease in the expression of miR-130a and miR-185 in MNCs from SEB+RES treated mice when compared to SEB+VEH treated group. Furthermore, there was an inverse relationship between miR-130a and miR-185 with CSF-1 gene expression in liver MNCs. The data presented demonstrate for the first time that RES can effectively attenuate SEB-induced ALI by inducing the proliferation of MDSC. In summary, the beneficial effects of RES may be mediated by changes in miRs regulating pathways leading to the generation of immunosuppressive cells and/or production of anti-inflammatory cytokines suppressing SEB-induced ALI.
p38 MAPK is critical for nuclear translocation of IRF-7 during CpG-induced type I IFN expression in human plasmacytoid dendritic cells
Authors: Wang Qi , Reszka-Blanco Natalia , Cheng Liang , Li Guangming , Zhang Liguo , Su Lishan 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDCs) are the major source of type I IFN (IFN-I) in response to viral and bacterial infection that are recognized through the Toll-like receptor 7 (TLR7) or TLR9 signaling pathway. TLR9 agonist, oligodeoxyribonucleotides containing unmethylated CpG motifs (CpG), stimulates pDC to produce IFN-I but the mechanism keeps incompletely defined. Here, we established a human pDC cell line with the gLuc under control of the IFNa4 promtoer/enhancer, and utilized the high throughput assay to screen for protein kinase inhibitors to identify small molecules targeting IFN-I induction pathway in pDCs. We identified p38 MAPK inhibitors that show ability to inhibit IFN-I expression in pDCs without cytotoxicity upon CpG stimulation. p38 MAPK inhibitor inhibits IFN-I expression in a dose-dependent fashion in vitro and significantly suppressed IFN-I expression in humanized mice upon CpG treatment. There was a dramatic defect in the nuclear translocation of IFN regulatory factor 7 (IRF-7) in the prescence of p38 MAPK inhibitor. Therefore, p38 MAPK enhances IRF-7 nuclear translocation and IFN-I expression in human pDCs. These findings identify p38 MAPK as a target for modulating pDC activity.
Tuning of TCR signaling pathways in chronically stimulated CD4+ T cells in vivo
Authors: Matson Courtney Alexandra , Singh Nevil J. 
Published: January 2023

Abstract
Prolonged antigenic stimulation is known to tune down a T cell’s ability to respond to antigen. It is not yet fully understood at what level this tuning occurs. Using an adoptive transfer model where transgenic T cells are continuously stimulated by a persistent self-antigen in vivo, we have previously found that tuned T cells show poor responses to antigen-restimulation; however, stimulation with PMA and ionomycin is mostly intact. This implies the negative regulation during tuning operates mostly on the TCR proximal signaling machinery; upstream of PKC activation and Ca2+ flux. Ex vivo, tuned T cells show decreased Zap70 kinase activity. Additionally, downstream of Zap70, chronically stimulated cells have reduced expression of both c-Fos and c-Jun leading to decreased assembly of the AP-1 transcriptional complex. Finally, chronically stimulated cells have increased expression of IκBβ, which contributes to a decrease in NFκB signaling. An important characteristic of T cells in this model is that the loss of responsiveness is mostly reversible. Tuned T cells, when adoptively transferred into an antigen-free host recover most of their functionality and can re-respond to antigen. However, acute antigen removal for at least 48 hours in vitro retains an unresponsive state suggesting the negative regulation is stable for a significant period after removal from antigen. Importantly, the stability of tuning ex vivo allows us to screen for pharmacological agents capable of improving antigen responsiveness in these T cells. Such agents may prove therapeutic for increasing T cell responses in cancer and chronic infections.
Platelet activating factor as a novel danger signal for activation of NLRP3 inflammasome
Authors: Deng Meng , Brickey W. June , Guo Haitao , Tam Jason W. , Johnson Brandon M. , New J. Stewart , Koller Beverly H. , Kearney John F. , Ting Jenny P.-Y. 
Published: January 2023

Abstract
Platelet activating factor (PAF) is a biologically active phospholipid that promotes pathophysiological inflammation through binding to a unique G-protein-coupled seven transmembrane receptor (PAFR). Increased PAF level has been correlated with a number of inflammatory disorders, yet the molecular mechanism underlying the pro-inflammatory function of PAF remains incompletely elucidated. Here we showed PAF activates canonical NLRP3 inflammasome, resulting in ASC oligomerization, caspase-1 processing, and IL-1β and IL-18 maturation. PAF-induced IL-1β maturation is abrogated in macrophage from Nlrp3−/−, Asc−/−, Caspase-1−/− mice, but not Nlrc4−/−, Nlrp1−/−, Aim2−/−, Caspase-11−/− mice. In addition, NEK7, a new component of the NLRP3 inflammasome, is also essential for PAF induced IL-1β maturation. PAF activation of the NLRP3 inflammasome is dependent on calcium influx, potassium efflux, but independent of lysosome cathepsin, mitochondrial reactive oxygen species (ROS), and necroptosis. Surprisingly, PAFR is not essential for PAF induced NLRP3 inflammasome activation. Physiologically, PAF induced shock is protected in NLRP3 inflammasome component deficient mice. Together, our findings reveal a new PAFR independent pathway for detecting PAF which represents an unprecedented danger signaling function of PAF, and provide new insight for understanding the pathogenesis of PAF-mediated inflammatory disorders.
The Heat Shock Protein, gp96, initiates inflammatory responses by activation of inflammasome signaling platforms in macrophages
Authors: Wang Yifei , Binder Robert J. 
Published: January 2023

Abstract
Several members of the heat shock protein family elicit immune responses which are, in part, a result of activation of antigen presenting cells. Activated antigen presenting cells respond to these immunogenic HSPs by up-regulating expression of co-stimulatory molecules and secretion of a panel of pro-inflammatory cytokines including IL-1b. We have tested here the requirement for activation of inflammasomes in the release of IL-1b by one immunogenic HSP, gp96. Our results show that macrophages activate NLRP3 inflammasomes in response to gp96, by K+ efflux. Using a gp96 secretion system we show that these events initiate inflammatory conditions in mice, in the absence of additional known inflammasome activators or infection. These results are important because the document a novel mechanism by which proteins of endogenous origin, the HSPs, can modulate a sterile inflammatory response following their release from cells are involved in host protection against infections by regulating IL-1b secretion via inflammsome activation.
A natural polymorphism of EsxH in Mycobacterium tuberculosis disrupts the immunodominant CD8 T cell response.
Authors: Sutiwisesak Rujapak , Carpenter Stephen , Behar Samuel M. 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb), the cause of tuberculosis (TB), is an intracellular bacterium that evades antibacterial immunity. As control of Mtb requires T cell immunity, most vaccine strategies focus on eliciting protective T cells. How Mtb benefits from promoting T cell activation or evading T cell recognition is not certain. Here, we took advantage of a naturally occurring Mtb polymorphism to address this question in mouse model.
EsxH elicits an immunodominant CD8 T cell response. At the peak of immunity against Mtb in C57BL/6 mice, 30–50% of CD8 T cells are specific for EsxH4–11. Clinical Isolate 667 (CI667) is an Mtb strain containing a polymorphism in EsxH that has a single amino acid change of WT EsxH4–11 epitope from IMYNYPAM into IMYNYPTM (A10T). We hypothesized that this naturally occurring polymorphism would alter the hierarchy of CD8 T cell responses elicited by Mtb.
Following Erdman (WT) Mtb infection of C57BL/6 mice, CD8 T cells respond to three distinct epitopes, including the reported immunodominant EsxH4–11 epitope. In contrast, there was no response to any of EsxH epitopes after CI667 infection. Instead, the subdominant response to the 32C antigen became dominant. In BALB/c mice, CI667 stimulated a reduced but detectable CD8 response to EsxH20–28; the CD4 response to EsxH74–88 was similar to WT Mtb infection.
These results show that naturally occurring polymorphisms can have drastic effects on T cell immunity. We hypothesize that EsxH acts as a decoy antigen, and distracts the CD8 response from focusing on antigens that are presented by infected macrophages and protective. We are in the process of testing this hypothesis by developing isogenic Mtb strains that express EsxHErdman or EsxHCI667.
IFN-γ synergizes with polyI:C to reduce growth of murine and human cancer cell growth, with concomitant reductions in cyclinD1 and PD-L1.
Authors: Petro Thomas M. , Guinn Zachary P. 
Published: January 2023

Abstract
We saw that IFN-γ synergizes with poly I:C, a TLR3 agonist, in ISG54 expression. Interestingly, ISG54 induction correlated with reduced B16 melanoma growth. To study IFN-γ synergism with other PRR agonists, we treated B16 cells, as well as UM-SCC1 and UM-SCC38 human squamous cell carcinoma cells, with IFN-γ (20 ng/ml) with or without Loxoribine (Lox; TLR7), ODN1585 (TLR9), Zymosan (TLR2), cGAMP (STING), LPS (TLR4), or poly I:C (TLR3). IFN-γ or Lox alone reduced growth of each tumor cell type, poly I:C reduced growth of SCC1 cells, while Lox and poly I:C synergized with IFN-γ to reduce tumor cell growth. Moreover, SCCs required treatment with the combination of IFN- γ/poly I:C for 24 h to achieve growth cessation, while B16 cells required treatment for 48 h. IFN-γ alone induced ISG54, while no PRR agonist alone induced ISG54 in B16 cells, as determined by western blot. ISG54 was induced in SCC1 by poly I:C. All PRR agonists synergized with IFN-γ for greater levels of ISG54 in B16 cells. We also evaluated cyclin D1, required for cell cycle G1 progression, and p15, which inhibits G1 progression. IFN-γ or poly I:C alone had no effect on cyclin D1 or p15. However, IFN-γ plus poly I:C decreased cyclin D1 levels. We also evaluated PD-L1, an immune checkpoint protein, in B16 cells treated with IFN-γ ± polyI:C. IFN-γ or polyI:C alone had no effect PD-L1 levels in B16 cells, but the combination of IFN- γ plus poly I:C decreased PD-L1. Thus, IFN-γ plus polyI:C sharply reduce growth of mouse and human tumor cells, which is accompanied by reductions in Cyclin D1, which is involved in cell cycle progression, and PD-L1, which decreases antitumor immune responses. Thus IFN-γ plus polyI:C could be an effective adjuvant to cancer immunotherapy.
Molecular characterization of combination pattern recognition receptor agonist exposure on dendritic cell responses
Authors: Brown Deborah M. , Lampe Anna T. , Hain Jacey L. , Pannier Angela K. 
Published: January 2023

Abstract
Current Influenza A virus (IAV) vaccines induce antibody formation and therefore fail to induce heterosubtypic protection needed to overcome viral antigenic shift and drift. New vaccine strategies have used pattern recognition receptor (PRR) agonists as adjuvants to improve both B and T cell memory; however, the effect of using combined PRR agonists on vaccine induced memory is unknown. We have demonstrated that vaccination with hemagglutinin (HA) antigen with combined TLR4 and TLR9 agonists promotes protection from lethal infection more effectively than either agonist alone or HA with the oil-in-water adjuvant Addavax. To begin to dissect the molecular changes that occur in dendritic cells exposed to combination agonists, we used qPCR, multiplex assays and RNA-seq analysis to characterize the changes in gene expression in response to combined TLR4 and TLR9 stimulation. We found low dose TLR4 and TLR9 agonists in combination could enhance IL-6, IFN-β and CCL4 mRNA over either agonist alone. Analysis of in vivo early cytokine responses 6 hours post intramuscular vaccination, corroborated in vitro results, demonstrating increased levels of serum IL-6, TNF-α, CCL4 and CCL2 in response to combined agonists. To further probe the mechanistic basis for increased protection with combined agonists, transcriptome analysis was performed on dendritic cells after treatment with single or combination TLR4 and TLR9 agonists. Genes such as CXCL10, IL-12 and a number of interferon stimulated genes (ISGs) were shown to be increased following combination agonist stimulation. Further, gene ontology analysis and ingenuity pathway analysis will be performed to gain insight into the complex regulation and mechanism of dual TLR4 and TLR9 agonists.
Mucosal immunity and protective efficacy of inactivated influenza virus vaccine is improved by intranasal chitosan nanoparticle delivery in pigs
Authors: Dhakal Santosh , Renu Sankar , Ghimire Shristi , Lakshmanappa Yashavanth Shaan , Hogshead Bradley T. , Ruiz Ninoshkaly F. , Krakowka Steven , Lee Chang-Won , Gourapura Renukaradhya J. 
Published: January 2023

Abstract
Currently used inactivated influenza virus vaccines administered by intramuscular injection provide homologous protection, but the much required heterologous protection against constantly evolving field viruses is limited, attributed to lack of induction of mucosal IgA and cellular immune responses in the respiratory tract. We developed a novel vaccine delivery platform by encapsulating swine influenza A virus (SwIAV) killed/inactivated antigen (KAg) in mucoadhesive, biodegradable, polycationic chitosan nanoparticles (CNPs-KAg). The CNPs-KAg vaccine (&lt;500nm in size) was administered through intranasal route as mist to nursery pigs twice and challenged with a zoonotic and virulent heterologous SwIAV. Pigs vaccinated with CNPs-KAg had enhanced mucosal IgA response throughout the respiratory tract against homologous (H1N2), heterologous (H1N1) and heterosubtypic (H3N2) SwIAV. In addition, the frequency of cytotoxic T cells and T helper/memory cells were increased in CNPs-KAg vaccinated pigs, together with increased level of systemic and mucosal recall IFNγ response. In heterologous virus challenged pigs, CNPs-KAg vaccine reduced the pneumonic lesions and virus titers in lungs and significantly reduced the nasal viral shedding. In conclusion, intranasal delivery of chitosan nanovaccine elicited better cross-protective mucosal IgA and cellular immune responses in the respiratory tract and reduced viral load and shedding, indicating that chitosan based influenza nanovaccine might be an ideal candidate vaccine against SwIAV infections under field conditions.
Metabolic activity as a surrogate marker of immune cell activation following vaccination
Authors: Waickman Adam T. , Friberg Heather , Jarman Richard , Currier Jeffrey R. 
Published: January 2023

Abstract
The identification and quantification of lymphocytes activated in response to vaccination is a critical first step in characterizing the efficacy of any candidate vaccine platform. Traditionally, this has been accomplished by monitoring the upregulation of defined activation markers on circulating lymphocytes by flow cytometry, or the measuring the production of effector cytokines following ex vivo stimulation. However, there is significant variability in both the kinetics and magnitude of activation marker up-regulation or cytokine production following stimulation even within the same type of lymphocyte. Therefore, traditional methodologies for identifying activated lymphocytes may offer a significantly skewed perspective on exactly which immune cells are responding to vaccination or natural infection. Herein, we examine the utility of measuring metabolic activity of circulating lymphocytes as a marker of activation following immunization with a live-attenuated dengue vaccine. Unlike any other marker, upregulation of metabolic activity is a universal indicator of lymphocyte activation which can be detected quickly after stimulation and persists throughout the functional stage of an immune response. We examine several methods of monitoring the metabolic state of both T cells and NK cells following immunization, and compare the size and diversity of the immune response generated relative to standard phenotyping protocols.
Immunomodulatory activity of 2-deoxyguanosine nanoparticles in DNA vaccines
Authors: Santos-Colón Saritza , Ramírez Maite , Miranda Eric , Reyes Nelson , Martinez Osmarie , Acosta Maxier , Rivera José , Otero Miguel 
Published: January 2023

Abstract
DNA vaccines induce protective immune responses against several pathogens and tumor cells, but strategies to improve their effectiveness are still under study. Only two adjuvants have been licensed by the FDA, which increases the needs for the discovery of novel adjuvants for HIV vaccines and other infectious diseases. In this regard, we study the immunomodulatory properties of 2-Deoxyguanosine-based nanoparticles (SHS) in DNA vaccines coding for HIV Gag (NL4-3 clone) and Vaccinia Virus Western Reserve (VVWR) A27 proteins. The sequence of A27L was codon optimized and linked to the Outer Domain-1 (OD1) of gp120 HIV-1 protein sequence serving as an additional molecular adjuvant. In this study, female Balb/c mice were immunized three times intramuscularly with plasmids (pGag and pOD1-A27L). Then, seven days after the last dose, cellular and humoral immunity were evaluated through ELISpot and ELISA assays. We found significantly higher frequencies of IFN-γ-producing cells in mouse immunized with pGag and pOD1-A27L co-formulated with SHS versus controls. We also found higher significant levels of IFN-γ over IL-4 in supernatant of splenocytes isolated from mice receiving pGag and pOD1-A27L combined with SHS. Moreover, administration of pOD1-A27L+SHS stimulated a significant higher production of total IgG in mice when compared to Naïve group. Here, we demonstrated that pGag and pOD1-A27L vaccines co-formulated with SHS nanoparticles successfully activate adaptive immunity through production of Th-1 cytokines and/or IgG antibodies. As this type of immunity is essential to clear many viral infections, these data support the evaluation of SHS nanoparticles as a novel adjuvant for DNA vaccines against multiple viral infectious agents.
Safety, tolerance, and effects on alveolar macrophages of a nebulized TLR agonist combination in foals
Authors: Bordin Angela Ilha , Bray Joselyne , Rocha Joana , Markesich Diane , Cohen Noah 
Published: January 2023

Abstract
The intracellular pathogen R. equi causes pneumonia in foals and people that closely resembles that caused by Mycobacterium tuberculosis (Mtb). Development of resistance to antimicrobials by R. equi and Mtb represents a major health problem for veterinary and human medicine. There is a need for control methods and treatment alternative to the use of antibiotics. Our alternative approach is to use a TLR2/6 and TLR9 agonist combination known as PUL-042 to stimulate innate immunity in the lungs. Our objectives were to evaluate safety and ex vivo efficacy of PUL-042 (Pulmotect Inc, Houston, TX) in newborn foals. Ten newborn foals were divided in 2 groups (PUL-042 and saline control) and submitted to a bronchoalveolar lavage (BAL) on day 2 of age, 6 nebulizations in a 2-week interval and a second BAL 24 h after the last nebulization. BAL cells were infected with a virulent strain of R. equi, and then lysed at either 0 h (T0) and after 48 h of culture (T48). Supernatants were saved for determination of INF-γ, IL-1β, and IL-10 concentration by ELISA. PUL-042 did not induce either adverse effects or airway inflammation in foals, as assessed by clinical examination and BAL cytology. At the tested doses, nebulization of PUL-042 did not significantly (P &gt; 0.05) affect ex vivo killing capacity of R. equi by AMs of foals at 2 and 21 days of age relative to controls. Results of cytokines are pending. Repeat nebulization of newborn foals with PUL-042 did not induce any adverse effects nor an inflammatory response in the lungs. At the dose tested, PUL-042 did not affect the killing capacity of R. equi by AMs, but its effects on other immune cells, and at other doses, need further evaluation.
ICOS-mediated phosphoinositide 3-kinase signaling controls induction and maintenance of collagen-induced arthritis
Authors: Panneton Vincent , Yazdchi Sahar Bagherzadeh , Witalis Mariko , Chang Jinsam , Suh Woong-Kyung 
Published: January 2023

Abstract
The inducible T-cell costimulator (ICOS) is a key costimulatory receptor facilitating differentiation and function of follicular helper T cells and inflammatory T cells. Rheumatoid arthritis patients were shown to have elevated levels of ICOS+ T cells in the synovial fluid suggesting a potential role of ICOS-mediated T cell costimulation in autoimmune joint inflammation. In this study, using ICOS knockout and knockin mouse models, we found that ICOS-mediated phosphoinositide 3-kinase (PI3K) signaling is required for the induction and maintenance of collagen-induced arthritis (CIA), a murine model of rheumatoid arthritis. For the initiation of CIA, ICOS-mediated PI3K signaling was critical for antibody production and expansion of inflammatory T cells. Further, we found that ICOS-PI3K signaling is important for maintenance of CIA in an antibody independent manner. Importantly, we found that a small molecule inhibitor of glycolysis, 3-bromopyruvate, ameliorates established CIA suggesting a potential link between ICOS-PI3K signaling and glucose metabolism. Thus, we identified ICOS-PI3K axis as a key signaling pathway that controls induction and maintenance of CIA and provide evidence that T cell glycolytic pathways can be potential therapeutic targets for rheumatoid arthritis.