bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autophagy sustains the survival and pro-tumorigenic effects of neutrophils in human cancer (P2053)
Authors: Li Xue-Feng , Kuang Dong-Ming , Zheng Limin 
Published: January 2023

Abstract
Neutrophils are a common inflammatory infiltrate that are enriched predominantly in the peritumoral stroma of many human cancers. The underlying mechanisms, however, that allow tumor microenvironments to educate neutrophils in peritumoral stroma are largely unknown. Herein we showed that most neutrophils in peritumoral stroma areas, but not those within the blood vessels, expressed significant higher amount of autophagy-related LC3 protein. In consistence, exposure of neutrophils to culture supernatants from several types of solid tumor cells (TSN) resulted in a rapid up-regulation of autophagy in cells, an effect that could be inhibited by blocking the activation of Erk1/2, p38 and NF-κB signaling pathways. The increased autophagy in TSN-exposed neutrophils was associated with preserved mitochondria integrity, reduced production of mitochondrial reactive species and cleavage of Caspase 3, and attenuated apoptosis in these cells. Pretreatment with 3-methyladenine, a specific autophagy inhibitor, could effectively induce mitochondrial injury and subsequent apoptosis in neutrophils. Moreover, we provided evidence that the induction of autophagy markedly enhance the pro-tumorigenic activities of neutrophils by releasing higher amount of matrix metalloproteinase-9, hepatocyte growth factor, and oncostatin M. Thus, increased autophagy in tumor-associated neutrophils may represent a novel mechanism that links the inflammatory response to neoplastic metastasis.
Increased B7-1 (CD80) expression reduces overall tumorigenicity and metastatic potential of the murine pancreatic cancer cell model Pan02 (P2085)
Authors: Imasuen Imade , Bozeman Erica , He Sara , Patel Jaina , Selvaraj Periasamy 
Published: January 2023

Abstract
Pancreatic cancer is both difficult to detect and harbors survival rates as low as 5%, thus heightening the need for better and new therapies. Several molecules have been investigated to increase the immunogenicity of tumor cells including the costimulatory molecule B7-1. The murine pancreatic cancer cell line, Pan02, expresses low, endogenous levels of B7-1; however, the role of B7-1 in pancreatic cancer progression has not yet been elucidated. We hypothesized that increased cell surface expression of B7-1 in the Pan02 model would decrease tumor growth and metastasis. To test this, Pan02 cells were transfected with mB7-1 (Pan02-mB7-1). Flow cytometry analysis confirmed a 40-fold increase in B7-1 cell surface expression in transfected cells compared to WT. In order to assess the in vivo effects of increased B7-1 expression, C57BL/6 mice, were challenged with Pan02-mB7-1 and Pan02-WT cells. We observed that B7-1 transfected Pan02 cells showed an overall decrease in tumor kinetics compared to mice challenged with Pan02-WT. In addition, increased cell surface B7-1 expression prevented metastasis to the lung in tumor positive C57BL-6 mice. More interestingly, Pan02 cells that metastasized to the lung (Pan02-RG-M-Lung) from WT challenged mice showed lower B7-1 expression by flow cytometry compared to Pan02-WT cells. Taken together, increased cell surface B7-1 expression is successful in inhibiting tumor growth and preventing metastasis in the murine pancreatic cancer model.
Impairment of cytotoxic anti-tumor response by hypoxic stress (P2011)
Authors: Chouaib Salem , Hasmim Meriem , Noman Zaeem M 
Published: January 2023

Abstract
Hypoxia is a central hallmark of tumor microenvironment and a pivotal driving force of malignant progression. Although hypoxia has been reported to play a major role in the acquisition of tumor resistance to cell death, the molecular mechanisms that enable the survival of hypoxic cancer cells have not been fully elucidated. Recently, attention has focused on the mechanisms by which hypoxic tumour cells alter their transcriptional profiles allowing them to persist under conditions of hypoxic stress. Here we demonstrate that NANOG, a transcription factor associated with stem cell self renewal, is induced under hypoxic conditions at both transcriptional and translational levels and contributes to the decreased susceptibility of target cells to specific cytotoxic T lymphocyte (CTL)-mediated lysis. The precise mechanisms by which NANOG regulates tumor susceptibility to CTL-mediated lysis will be discussed. In addition since we have demonstrated that in vivo inhibition of hypoxia restores cytotoxic T lymphocyte activity and promotes in vivo tumor regression, we will also discuss the relationship between hypoxic stress and survival pathways associated with hypoxic stress in particular the emergence of cancer stem cells and autophagy induction.
Evaluation of the protective immunity of a novel subunit fusion vaccine in a murine model of systemic MRSA infection (P4254)
Authors: Zeng Hao 
Published: January 2023

Abstract
Staphylococcus aureus is a common commensal organism in humans and a major cause of bacteremia and hospital acquired infection. Because of the spread of strains resistant to antibiotics, these infections are becoming more difficult to treat. Therefore, exploration of anti-staphylococcal vaccines is currently a high priority. Iron surface determinant B (IsdB) is an iron-regulated cell wall-anchored surface protein of S. aureus. Alpha-toxin (Hla) is a secreted cytolytic pore-forming toxin. Previous studies reported that immunization with IsdB or Hla protected animals against S. aureus infection. To develop a broadly protective vaccine, we constructed chimeric vaccines based on Hla and IsdB antigenic region. Immunization with the chimeric bivalent vaccine induced strong antibody and T cell responses. When the protective efficacy of the chimeric bivalent vaccine was compared to that of individual proteins in a murine model of systemic S. aureus infection, the bivalent vaccine showed a stronger protective immune response than the individual proteins (IsdB or Hla). our results verified that the strategy of optimized chimeric bivalent antigens could be as an important approach for developing S. aureus mult-subunit vaccines against S. aureus infection.
Local and systemic immune responses predict clinical outcomes in influenza virus infection independently of age and viral titer (P3000)
Authors: Oshansky Christine , Gartland Andrew , Wong Sook-San , Jeevan Trushar , Wang David , Roddam Philippa , Caniza Miguela , Hertz Tomer , DeVincenzo John , Webby Richard , Thomas Paul 
Published: January 2023

Abstract
Influenza infection can result in mild or serious illness in some individuals, and studies suggest that increased inflammatory mediators have a role in disease severity. To investigate the role of inflammation in clinical disease, we performed a prospective, longitudinal study, sampling symptoms, blood, nasal lavages, and viral loads from participants during natural influenza infection. Surprisingly, clinical disease defined by symptoms or hospitalization did not correlate with viral dynamics, but consistent with other clinical reports, age was associated with hospitalization. There was no association between age and viral dynamics, though a strong inverse correlation between age and cytokine levels in the nasal lavage was maintained even when adjusted for prior immunity, indicating that children mount more aggressive inflammatory responses independently of pathogen dynamics. Correcting for this age-association, analysis of cytokine levels in nasal lavages revealed that MCP3 and IFNα were predictors of respiratory symptoms, while plasma MCP3 and IL-10 predicted symptom severity and hospitalization. In all cases, associations were not dependent on viral load. The association between IL-10 and symptom severity was confirmed in an independent cohort. In summary, children suffer significantly higher local inflammation, and a small subset of cytokines, including MCP3 and IL-10, serve as diagnostic markers for clinical outcome of influenza infection in all age groups.
Detection of antigen-specific B cells to Plasmodium falciparum apical membrane antigen-1 and circumsporozoite protein in humans and immunized mice (P3025)
Authors: Odumade Oludare , Frosch Anne , Taylor Justin , Dehnbostel Joanne , John Chandy 
Published: January 2023

Abstract
Worldwide, there are an estimated 216 million cases of malaria annually and of those affected 655,000 will succumb to the disease. Antibody responses to infection have been shown to be critical in limiting infection and disease, but the mechanisms by which these are acquired or lost are poorly understood. Various hypotheses have been proposed for dysfunctional humoral responses including B cell exhaustion, germinal center architectural disruption, parasite antigenic variation and inhibitory innate immune signaling. However, the relative infrequency of malaria specific B cells in the periphery has limited targeted detection of these cells. We present a strategy for phenotypic analyses of malaria specific B cells in peripheral blood using tetramerized biotinylated malaria antigen coupled to phycoerythrin labeled streptavidin, magnetic bead enrichment and cytometric flow analysis. Using this technique, we demonstrate the detection of an antigen specific germinal center reaction in B6 mice after immunization with P.pastoris expressing recombinant Plasmodium falciparum apical membrane antigen-1 and circumsporozoite protein. Preliminary results in humans identified low frequency B cells specific to these two antigens. Overall, our results suggest that this technique will be a valuable tool in studying naturally acquired and vaccine derived malaria specific B cell responses.
CD16+ expressing monocyte subsets and CD14+ HIV DNA levels predict HIV-associated neurocognitive disorders (HAND) in treatment-naïve HIV infected Thai subjects (P3043)
Authors: Byron Mary Margaret , Valcour Victor , Ananworanich Jintanat , Agsalda Melissa , Chalermchai Thep , Tipsuk Somporn , Sithinamsuwan Pasiri , Schuetz Alexandra , Hutchings Nicholas , Barbour Jason , Phanuphak Nittaya , Shikuma Cecilia , Shiramizu Bruce , Ndhlovu Lishomwa 
Published: January 2023

Abstract
Since the advent of anti-retroviral (ARV) therapy, HIV-dementia has significantly declined, however, milder forms of HIV-associated Neurocognitive Disorder (HAND) remain highly prevalent. HIV DNA reservoirs in CD14+ Monocytes (MO) have been associated with cognitive impairment. However, the role of MO subsets in the pathogenesis of HAND over time remains poorly characterized. We sought to analyze HIV DNA content and MO phenotypic properties in relation to cognitive impairment through a prospective study of 30 ARV therapy-naïve individuals enrolled by HIV DNA reservoir levels with subsequent cognitive characterization as normal (n=17), Asymptomatic Neurocognitive Impairment (n=6), Mild Neurocognitive Disorder (MND) (n=2), HIV-associated dementia (HAD) (n=5) and 44 HIV- matched controls. HIV DNA was extracted from CD14+ cells and quantified by PCR. MO subsets and cell surface chemokine receptors were measured by flow cytometry using an extensive panel to exclude non-MO. Our preliminary findings show that among 4 MO subsets, there was a statistically significant decline in frequency of CD14highCD16+ (p&lt;0.001) and an increase in CD14lowCD16- (p&lt;0.001) in HIV+ compared to HIV- individuals. Remarkably, we found that CD14lowCD16+ MO frequency was lower in cognitively impaired individuals (p&lt;0.05) and frequency of CCR2+ and CCR5+ expressing MO was higher in symptomatic (MND and HAD) cognitively impaired HIV+ individuals (p&lt;0.05, p&lt;0.05).
RIP3 is involved in host defense against HSV-1 infection via enhancing STING mediated antiviral signaling (P1405)
Authors: Liang Yaoji , Wu Suqin , Han Jiahuai 
Published: January 2023

Abstract
Herpes simplex virus type 1 (HSV-1), a double stranded DNA virus, is a widespread virus that infects approximately 80% of adults worldwide. HSV-1 infection is also the major cause of fatal childhood HSV-1 encephalitis (HSE). The mechanisms of how host cells defend against HSV-1 are still not fully known. Here we report that receptor interacting protein 3 (RIP3) enhances type I interferon production in response to HSV-1 infection. In our study of the functions of RIP3, we have found that the HSV-1 replication in RIP3 knockout L929 was unrestrained, and that HSV-1 induced TBK1 and IRF3 activations were impaired in RIP3 deficient cells. Further study has shown that over-expression of wild type RIP3, but not RHIM-mutant RIP3, enhances type I interferon production, especially when co-expressed with STING, a central signaling molecule in the innate immune response to DNA virus infection. Upon treating the cell with HSV-1, TBK1 was recruited to RIP3. In addition, RIP3 can promote the interaction between STING and TBK1 in a RHIM dependent manner. We propose that upon HSV-1 stimulation, RIP3 promotes the recruitment of TBK1 to STING, which leads to TBK1-mediated IRF3 activation and subsequent type I IFN secretion. Our study revealed that, in addition to its function in necrosis, RIP3 plays a role in type I interferon production. These results may also provide a new perspective for the exploration of childhood HSE pathogenesis.
Unanchored Lysine48-linked polyubiquitin chains positively regulate the type I IFN-mediated antiviral response (P1391)
Authors: Rajsbaum Ricardo , Versteeg Gijs , Schmid Sonja , Maestre Ana , Belicha-Villanueva Alan , Fernandez-Sesma Ana , tenOever Benjamin , García-Sastre Adolfo 
Published: January 2023

Abstract
Type-I interferons (IFN-I) are essential antiviral cytokines produced upon microbial infection. IFN-I elicits this activity through the upregulation of 100s of IFN-I stimulated genes (ISGs), many of which have known antiviral activity. The full breadth of ISG induction requires activation of a number of cellular factors including the IκB kinase epsilon (IKKϵ). However, the mechanism of IKKϵ activation upon viral infection or IFN receptor signaling remains elusive. Here we show that TRIM6, a member of the E3-ubiquitin ligase tripartite motif (TRIM) family of proteins, interacts with IKKϵ and promotes induction of IKKϵ-dependent ISGs. Some studies have suggested a role for unanchored K63-linked polyubiquitin chains, which are not conjugated to any protein, in regulation of kinase activity. However, no role has yet been established for unanchored K48-linked polyubiquitin chains in kinase activation. We show that TRIM6 and the E2-ubiquitin conjugase UbE2K cooperate in the synthesis of unanchored K48-linked polyubiquitin chains, which activate IKKϵ for subsequent STAT1 phosphorylation. Our work defines a previously unrecognized activating role of K48-linked unanchored polyubiquitin chains in kinase activation and identifies the UbE2K-TRIM6-ubiquitin axis as critical for IFN signaling and antiviral response.
The role of IFI16 in the innate immune response to cytosolic nucleic acids (P1395)
Authors: Thompson Mikayla , Jensen Soren , Sharma Shruti , Fitzgerald Katherine , Kurt-Jones Evelyn 
Published: January 2023

Abstract
Recently, the Interferon Gamma Inducible (IFI)16 was linked to cytoplasmic DNA induced type I interferon (IFN) responses. IFI16 was shown to associate with Stimulator of IFN genes (STING), leading to TANK binding kinase-1 (TBK1) dependent phosphorylation of interferon regulatory factor 3 (IRF3) and transcription of type I IFN genes. Here we created THP1 monocytic cell lines with stable knockdown of IFI16 by lentiviral transduction of shRNA and demonstrated that IFI16 knockdown leads to a severely attenuated type I IFN response following treatment of cells with either synthetic DNA ligands, or after infection with DNA viruses such as HSV-1. Surprisingly, these cells were also compromised in their ability to induce type I IFNs to other non-DNA ligands including synthetic 5’pppRNA and Sendai Virus, both of which signal via RIG-I. In contrast the NF-kB regulated cytokines IL-6 and IL-1β remain unaffected in IFI16 knockdown cells. Utilizing Nanostring technology, we also determined that IFI16 is capable of regulating DNA and RNA induced expression of a panel of Interferon Stimulated genes (ISGs), including RIG-I. We also monitored key steps in the type 1 IFN pathway and found that levels of total IRF3 were also reduced in IFI16 knockdown cells. These results suggest that IFI16 senses DNA viruses, eliciting a type I IFN response, but also acts more broadly in the regulation of ISGs in response to both DNA and RNA viruses.
Integrin α9 on lymphatic endothelial cell at draining lymph node regulates lymphocyte egress via controlling S1P secretion during inflammation (P5114)
Authors: Ito Koyu , Morimoto Junko , Kihara Akio , Simmons Szandor , Ishii Masaru , Uede Toshimitsu 
Published: January 2023

Abstract
Sphingosine 1-phosphate (S1P) plays a role in lymphocyte egress from lymphoid organs. However, it remains unclear how S1P production and secretion are regulated. We found that α9 integrin and its ligand, Tenascin-C (TN-C) were co-localized on lymphatic endothelial cell (LEC) of medullary sinus in the draining lymph nodes (dLN). To examine the role of α9 integrin in the dLN, mice that had been treated with Complete Freund Adjuvant (CFA) were treated with blocking anti-α9 integrin antibody (55A2C). Although CFA-induced inflammation was associated with appearance of migrating lymphocytes within medullary sinus at dLN, mice-treated with 55A2C showed vacant medullary sinus. In addition, adoptive T cell transfer experiments demonstrated that lymphocyte egress was significantly reduced by both FTY720, which was S1P receptor agonist, and 55A2C under inflamed condition. Stimulation of LEC by TN-C enhanced S1P secretion without affecting S1P secretion and / or degradation. We further show that blockade of α9 integrin-mediated signaling reduces lymphocyte egress from dLN in several models, including experimental autoimmune encephalomyelitis (EAE) where it improves clinical scores and pathology. Therefore, we propose that interaction of α9 integrin on LEC with TN-C induces S1P secretion, and subsequent lymphocyte egress, possibly by induction of S1P transporter and that manipulating α9 integrin function may offer a therapeutic strategy for treating various inflammatory disorders.
Conditional ablation of Langerhans cells exacerbates autoimmune corneal inflammation (P5134)
Authors: Gutierrez Miguel-Angel , Zhu Zejin , Philips Rachael , Eriksson Anna U , Halder Ramesh , Kim Peter , King Jennifer , Rao Narsing , Singh Ram Raj 
Published: January 2023

Abstract
Ocular surface inflammation is common and sometimes severe in autoimmune diseases. Little is known about the role of cornea-resident dendritic cells, including Langerhans cells (LC), in ocular surface autoimmunity. Here, we analyzed corneal LC in autoimmune-prone MRL mice. LC (CD11c+ CD207+) were more abundant and activated (CD86+ CD40+) in the corneal epithelium of MRL mice than of B6 mice. However, LC were ~5-fold lower in the corneal stroma of MRL mice than of B6 mice. LC in cervical lymph nodes were also lower in MRL mice than in B6 mice. Ongoing studies will examine whether the increase of activated LC in corneal epithelium, but their reduction in corneal stroma and eye-draining lymph nodes in autoimmune mice is due to a defect in the migration of LC from the cornea to eye-draining lymph nodes. Since LC are believed to carry antigens from tissues to their respective draining lymph nodes to maintain tolerance in a tissue-specific manner, the reduced LC in eye-draining lymph nodes may lead to the breakdown of tolerance to eye antigens. To directly test the role of LC in ocular autoimmunity, we introgressed the Lang-eGFP.DTR knockin mutation from the stock B6 mice that express diphtheria toxin receptor (DTR) driven by Langerin promoter (Malissen, 2005) onto the MRL background. Diphtheria toxin injections depleted LC in Lang-eGFP.DTR MRL mice and accelerated corneal inflammation. Taken together, these data suggest a protective role of LC in corneal inflammation.
Highly cytokinergic SPE-7 anti-dinitrophenyl-immunoglobulin E induces antigen-dependent thermal hyperalgesia and inflammatory cytokine production in the mouse hindpaw (P6000)
Authors: Chatterjea Devavani , Mack Madison , Wetzel Abigail , Tonc Elena , Balsells Evelyn , Engblom Camilla 
Published: January 2023

Abstract
SPE-7 anti-DNP-IgE initiates inflammatory cytokine production without antigen stimulation in vitro. In contrast, ϵ2624 anti-DNP IgE does not induce inflammatory cytokines independent of antigen-stimulation. However, their effects in vivo have not been described. We tested two well-characterized anti-mouse IgE antibody clones in our model of murine thermal hyperalgesia. We found pronounced hindpaw hyperalgesic responses lasting to at least 6 hours following antigen challenge after sensitization with SPE-7. These responses were accompanied by histamine release and neutrophil influx as we have previously shown in c48/80-mediated thermal hyperalgesia. In contrast, we found that antigen challenge following sensitization with clone ϵ2624 produced slight hyperalgesia at 1 hr that was resolved by 3 hours. We demonstrate a novel functional, behavioral difference between reactions mediated by these IgE antibody clones in vivo in mice. Also, SPE-7 does not lead to antigen-independent cytokine production in the mouse hind paw. After antigen challenge, both IgEs produce equivalent levels of TNF-α, IL-1, and IL-6, and cause equivalent histamine release and neutrophil influx. Thus tissue-resident sentinel mast cells may be critical initiators of allergen-evoked hyperalgesia in mice and differences in the nature and repertoire of IgE antibodies produced during an allergic response likely have pathophysiological implications for various health outcomes including pain.
Antibodies binding to discrete segments of CϵmX of mIgE on B lymphocytes (P6004)
Authors: Chan Yueh-Hsuan , Chu Hsing-Mao , Chen Jiun-Bo , Wu C. , Pan Chi-Chun , Chang Tse Wen 
Published: January 2023

Abstract
CϵmX is a discrete domain of 52 a.a. residues, located between the CH4 domain and the C-terminal membrane anchor peptide of the ϵ heavy chain of membrane-bound IgE (mIgE) on human B cells. Among the monoclonal antibodies (mAbs) prepared for their ability to bind to CϵmX fragments or CϵmX-containing recombinant proteins in ELISA, those that can bind to native mIgE on B cells can cause ADCC, apoptosis, and other cytolytic mechanisms on the B cells and are potential useful for controlling IgE production for treating allergic diseases. The CϵmX-specific mAbs prepared previously appear to recognize discrete and overlapping peptide segments of CϵmX. A linear peptide epitope of an antibody is defined by a contiguous peptide segment of minimal stretch that confers maximum binding by the antibody. In some instances, while the peptide epitopes recognized by two mAbs may be adjacent to each other and overlap, the two mAbs do not bind to the adjacent epitopes by a substantial degree. This can be illustrated by the pair of humanized mAbs, h4B12 and h47H4. The mAb h4B12 binds to peptide P1-15 (GLAGGSAQSQRAPDR) and h47H4 binds to peptide P7-18 (AQSQRAPDRVLC). Although the two epitopes share a segment AQSQRAPDR, the mAb h4B12 does not bind to P7-18, and h47H4 does not bind to P1-15. A related presentation from our group (Hsing-Mao Chu et al.) will also reveal that the shared peptide segment AQSQRAPDR has two markedly different conformations when it is associated with h4B12 or with h47H4.