bluuebunny/crossref_metadata_2025_split huggingface dataset:
Deletion of OLFM4 rescues defective host defense against Staphylococcus aureus, but not Aspergillus fumigatus in murine X-linked chronic granulomatous disease (P1259)
Authors: Liu Wenli , Sugui Jancy , Li Hongzhen , Kwon-Chung Kyung , Rodgers Griffin 
Published: January 2023

Abstract
Chronic granulomatous disease (CGD) patients have recurrent life-threating bacterial and fungal infections due to the mutation of one of four subunits of the respiratory burst oxidase (NADPH-oxidase). Olfactomedin 4 (OLFM4) is a neutrophil granule protein, which has been recently identified as a negative regulator of host innate immunity against bacteria infection in mice through modulation of neutrophil protease activity. The goal of this study was to evaluate the impact of OLFM4 deletion on host innate immunity against Staphylococcus aureus and Aspergillus fumigatus, in a murine X-linked CGD model. We found that neutrophil intracellular killing and in vivo clearance of S. aureus have been significantly increased in gp91phox- and OLFM4-double deficiency mice compared with CGD mice. The mice survival to S. aureus sepsis in gp91phox- and OLFM4-double deficiency mice has also been significantly prolonged compared with CGD mice. Cathepsin C activities in OLFM4 deficient as well as double deficient mice neutrophils were significantly higher than those in WT mouse neutrophils. Accordingly, the elastase and cathepsin G activities in the neutrophils of OLFM4 deficient and double deficient mice were also substantially higher than those in WT mice as well as CGD mice. However, we have not observed enhanced innate immunity against A. fumigatus in OLFM4 deficiency mice. OLFM4 might prove to be an important target in CGD patients to augment host defense against bacterial infection.
Systems biology analysis of the ceca and bursa revealed unique innate immune responses to Campylobacter jejuni in two genetically distinct lines of chickens (P1269)
Authors: Zhou Huaijun , Abernathy Jason , Nazmi Ali , Li Xianyao , Swaggerty Christina , Pevzner Igal , Drake Kenneth , Kogut Michael 
Published: January 2023

Abstract
Campylobacter jejuni (C. jejuni) is one of the most commonly reported causes of foodborne illness worldwide, although C. jejuni is a commensal in chickens. Two genetically distinct broiler lines, either resistant (line A) or susceptible (line B) to C. jejuni colonization, , were used to understand innate immune response to C. jejuni in a time-course (1, 4, 24, and 36 h post-inoculation) study. A total of 128 1-d-old chicks (64 per line), were either orally challenged with C. jejuni or inoculated with PBS. Total RNA were isolated from ceca and bursa, and a chicken 44K Agilent microarray was performed on all samples. Microarray data was then analyzed using a dynamic Bayesian network machine learning approach. Bursa samples from infected line A birds showed significantly more perturbations in carbohydrate and lipid metabolism compared to non-infected controls while susceptible line B birds responded with perturbations in signal transduction and amino acid biosynthesis. Differences were also observed in the ceca where apoptosis, MAPK and Toll-like receptor signaling pathways were significantly perturbed in line A birds while cell adhesion molecules, Jak-STAT signaling and fatty acid metabolism were significantly perturbed in line B birds. The distinct perturbed signaling pathways between these two lines and between two tissues provide new insights into the early immune response of chickens to C.jejuni and will allow for in silico hypothesis testing for future functional studies.
The role of miRNA in regulating bacterial clearance (P1273)
Authors: Tay Hock , Kaiko Gerard , Hansbro Philip , Foster Paul 
Published: January 2023

Abstract
RATIONALE MicroRNAs (miRNAs) are small non-coding RNA that can bind to multiple target mRNA to repress protein production. While the expressions of specific miRNAs are known to be dysregulated in multiple human diseases, the role of miRNAs in regulating bacterial infection is unclear. Our aim is to investigate the roles of miRNA in regulating these processes. METHODS BALB/c mice were challenged with low doses of NTHi. Lung miRNAs that were differentially expressed were identified by microarray. miRNA inhibitors (antagomirs) were used to knockdown miRNA expression to investigate the roles of miRNA in bacterial clearance in vitro and in vivo. RESULTS Upon NTHi infection, 20 miRNAs were up-regulated while 52 were down-regulated by &gt; 2.5 fold. Interestingly, knockdown of a specific miRNA in vitro decreased bacterial load and increased phagocytosis by macrophages and neutrophils. Similarly, transfer of macrophages or neutrophils deficient in miRNA into naïve recipient mice leads to enhance clearance of the bacteria in vivo. This enhance clearance is independent of cellular infiltration. Inhibiting miRNA may play a protective role by enhancing the function of innate immune cells during bacterial infection. CONCLUSION Our study suggests that miRNA may play important roles in regulating the innate immune response to bacterial infection.
Gene expression of SOAT1, a key enzyme for cholesteryl ester biosynthesis, is regulated by LPS and correlates with the antibacterial activity of human bronchial epithelial cell secretions against Pseudomonas aeruginosa (P1249)
Authors: Bahrami Bita , Anouseyan Rabin , Glavatska Maryna , Escobar Oswaldo , Porter Edith 
Published: January 2023

Abstract
Airway epithelial cells contribute to the first line of mucosal defense by secretion of antimicrobial peptides and antimicrobial lipids, including the cholesteryl ester (CE) cholesteryl arachidonate. Sterol O-acyltransferase 1 (SOAT1) is a key enzyme for CE biosynthesis. However, little is known about the regulation of the lipid-mediated arm of innate host defense. Polarized human bronchial epithelial cells (HBE) were incubated for 6 h with 100 ng/mL LPS, 10 μg/mL peptidoglycan (PG), 25 μg/mL PamCys3, and solvent control. SOAT1 mRNA was quantified by qRT-PCR with duplexing and RPLP0α as housekeeping gene. Lipid content of apical HBE secretions was analyzed by rpHPLC. SOAT1 expression was blocked with siRNA/lipofection. Antibacterial activity of HBE secretions was assessed by a 3 h colony forming unit assay with Pseudomonas aeruginosa (PA). SOAT1 gene expression was significantly increased by LPS (14.6 ±11.4, means ± SD, n=4, p=0.037). Stimulation with PG and PamCys3 increased SOAT1 expression by ~ 4- and 2-fold, respectively. Secretions from LPS-stimulated HBE showed an increase of total lipids including CA by ~30-50%. Selective inhibition of SOAT1 mRNA expression by 80% reduced the antibacterial activity of HBE whereby PA showed enhanced bacterial growth reaching &gt;170% of the controls. These data suggest that SOAT1 is regulated by conserved innate immunity pathways and further support the role of antimicrobial lipids as effector molecules in mucosal airway defense.
Induction of human Beta-Defensin-1 gene expression is dependent on IRF-7 and PU.1. (P1379)
Authors: Ryan Lisa , Dai Jihong , Fitzgerald-Bocarsly Patricia , Schwartz Kyell , Diamond Gill 
Published: January 2023

Abstract
Beta-defensins have been gaining recognition as immunomodulators and regulators of the inflammatory response. Human beta-defensin (hBD) peptides 1, 2 and 3 are differentially modulated in primary gingival and tracheal epithelial cells (EC) by viruses. We showed that both HSV-1, Sendai and PR8 influenza viruses increase the gene expression of hBD-1 in monocytes and plasmacytoid dendritic cells (PDC) and suppress gene expression of hBD-1 in target EC. Deleting the hBD-1 homologue in mice also increased influenza-induced morbidity, mortality and lung inflammation compared with wild type C57BL/6 mice, highlighting its importance in viral innate immunity. In this study, we investigated how hBD-1 gene expression is regulated. In PDC, no increased hBD-1 gene expression occurred when cells were incubated with the TLR9 agonist, CpG-A, indicating that hBD-1 is not induced via TLR9. When PDC were incubated with anti-interferon (IFN)-alpha-receptor-1 antibody, no change in viral induction by HSV-1 occurred. In addition, in a luciferase reporter system in A549 cells co-transfected with expression vectors containing IRF3, IRF5, IRF7 and PU.1, IRF7 and PU.1 were the only transcription factors that increased activation of the hBD-1 promoter. These results suggest that HSV-1 induction of hBD-1 occurs independently of IFN-alpha at the receptor level but that transcription factors common to IFN-alpha are involved in the regulation of hBD-1 gene expression.
EBI2 regulates type I IFN responses to TLR stimulation (P1409)
Authors: Chiang Eugene , Johnston Robert , Grogan Jane 
Published: January 2023

Abstract
Epstein-Barr virus induced receptor 2 (EBI2), a G protein-coupled receptor that has been shown to have important roles in segregating B cells within follicles, has recently been implicated as a regulator of an interferon regulatory factor 7 (IRF7)-driven inflammatory network (IDIN) that is associated with type I diabetes. Using plasmacytoid dendritic cells (pDCs) and CD11b+ myeloid cells from knockout mice deficient in EBI2, elevated production of type I IFN and IFN-α-dependent proinflammatory cytokines, respectively, were observed in response to in vitro stimulation with various TLR agonists. EBI2 appears to act as a negative regulator of IRF7-mediated type I IFN and IDIN in pDCs, as constitutive expression of IRF7 and IDIN genes were higher in EBI2 KO pDCs, likely accounting for the heightened primed state of responsiveness to endosomal TLR stimulation. Similar to its role on B cells, EBI2 plays a role in pDC localization. EBI2 KO pDCs failed to migrate in response to the EBI2 ligand 7α,25-dihydroxycholesterol and EBI2 KO mice had significantly lower frequencies of pDCs in the spleen and peritoneal cavity. Together, dysregulated migration of pDCs and elevated type I IFN responsiveness may be responsible for increased levels of IFN-α and IFN-β detected in the peritoneum of EBI2 KO mice following in vivo challenge with endosomal TLR agonists. These findings support a role for EBI2 as an important regulator of pDC and myeloid cell responses.
HSV-1 serine/threonine kinase US3 hyperphosphorylates IRF3 and inhibits the IFN-β production (P1413)
Authors: Zheng Chunfu , Wang Shuai 
Published: January 2023

Abstract
Viruses have evolved a variety of strategies to modulate host IFN-mediated immune responses. HSV-1 US3, a Ser/Thr kinase conserved in α-herpesviruses, has been demonstrated to be involved in multiple functions during HSV-1 infection. In this study, we reported the novel observation that US3 significantly inhibited Sendai virus (SeV) mediated activation of the IFN-β and IFN stimulated response elements (ISRE) promoters and the transcription of IFN-β, ISG54 and ISG56. US3 was also shown to block SeV-induced dimerization and nuclear translocation of IRF3. This inhibitory function required the kinase activity of US3, as kinase dead (KD) US3 mutants K220M and D305A could not inhibit the IFN-β production. US3 also inhibited IRF-3-responsive PRD(III-I) promoter activity induced by ectopic expression of the N-terminus of RIG-I (RIG-IN), MAVS, TBK-1, IKKϵ and IRF3/5D, indicating that US3 may block the IFN-β pathway at the IRF3 level. US3 was demonstrated to interact with and hyperphosphorylate IRF3 to prevent IRF3 activation. Finally, we constructed the US3 kinase dead mutant viruses, denoted as K220M or D305A HSV-1 respectively, and infection by both mutant viruses resulted in remarkable increase of IFN-β production compared with wild-type virus. To our knowledge, this is the first report that IRF3 serves as a substrate of HSV-1 Us3. These findings provide convincing evidence that US3 hyperphosphorylates IRF3, blocks the production of IFN-β, and subverts of the host innate immunity.
Inflammatory disease and impaired antiviral immunity due to unbalanced NF-kB and IRF3 activation result from a de novo human NEMO mutation (P1415)
Authors: Wessel Alex , Hsu Amy , Zilberman-Rudenko Jevgenia , Goldbach-Mansky Raphaela , Siegel Richard , Hanson Eric 
Published: January 2023

Abstract
The RIG-I-like receptors mediate antiviral signaling through NEMO-dependent activation of transcription factors NF-kB and IRF3, which are recognized for their roles in inflammation and the induction of Type I Interferon, respectively. We identified an individual with increased susceptibility to viral infection and persistent inflammatory disease. Sanger sequencing revealed a de novo synonymous mutation in the gene encoding NEMO that results in an in-frame splicing event and exclusive production of a shortened protein. To understand the role of NEMO in regulating the antiviral response, we stimulated patient and healthy control fibroblasts with RIG-I-like receptor ligand poly(I:C), which mimics viral RNA. Induction of IRF3 response genes IFNB1 and CXCL10 was reduced 35-fold and 500-fold (n=2), respectively, compared to healthy control, and patient cells were unable to control viral infection. In contrast, TNF induced the expression of proinflammatory genes TNFSF10 and IL17 7-fold and 4-fold higher in patient cells than in healthy control cells. Increased TNF induced gene expression was accompanied by increased p65 nuclear translocation indicative of increased canonical IKK activation. These results suggest that aberrant expression of an alternately spliced NEMO isoform constitutes a novel autoinflammatory disease characterized by unbalanced NF-kB and IRF3 mediated responses to viral infection.
Novel function of Trim44 promotes an antiviral response by stabilizing VISA (P1417)
Authors: Sun Bing , Yang Bo 
Published: January 2023

Abstract
VISA functions as a critical adaptor in the regulation of both the production of type I interferons (IFNs) and the subsequent control of the innate antiviral response. In this report, we demonstrate that Trim44 interacts with VISA and promotes VISA-mediated antiviral responses. The overexpression of Trim44 enhances the cellular response to viral infection, whereas Trim44 knockdown yields the opposite effect. Trim44 stabilizes VISA by preventing VISA ubiquitination and degradation. These findings suggest that Trim44 functions as a positive regulator of the virus-triggered immune response by enhancing the stability of VISA.
Intrinsic upregulation of critical innate immune effectors in a novel transgenic mouse model confers viral resistance (P1389)
Authors: Painter Meghan , Rodriguez Moses 
Published: January 2023

Abstract
Our laboratory generated a transgenic mouse that ubiquitously expresses 3D, a picorno viral RNA-dependent RNA polymerase. 3D transgenic mice display an extreme antiviral phenotype when challenged with viruses from distinct families. To probe this protective molecular mechanism, we assayed differential gene expression in tissues of uninfected 3D transgenic mice. Microarray analysis revealed increased levels of 78 genes &gt;4-fold, including a distinct upregulation of critical innate antiviral effectors across tissues. RT-PCR analysis of liver, lung, kidney, CNS and heart tissue from uninfected 3D mice showed a significant upregulation of antiviral genes (up to 300-fold). Following infection with EMCV, 3D transgenic mice exhibited significantly decreased viral tissue titers and survived lethal viral infection. This effect is specific to the 3D transgene since other mice expressing the picorno viral transgenes VP1 or VP2 did not display an antiviral phenotype. Studies of 3D-Ifnar1-/- or 3D-Ifngr1-/- mice, supported the conclusion that type I IFN signaling is necessary for the antiviral state in 3D mice, whereas IFNg signaling is dispensable. Maintenance of a viral-resistant state in uninfected 3D mice is independent of adaptive immunity since uninfected 3D-Rag1-/- mice are able to highly upregulate antiviral factors. Collectively, our data indicate that intrinsic upregulation of critical innate immune effectors confers broad-spectrum viral resistance in 3D mice.
STING recognition of cytoplasmic DNA instigates cellular defense (P1393)
Authors: Xia Tianli , Abe Takayuki , Harashima Ai , Konno Hiroyasu , Konno Keiko , Morales Alejo , Ahn Jeonghyun , Gutman Delia , Barber Glen 
Published: January 2023

Abstract
How the cell recognizes cytosolic DNA including DNA based microbes to trigger host defense related gene activation remains to be fully resolved. Here, we demonstrate that STING (for Stimulator of Interferon Genes), an endoplasmic reticulum (ER) translocon associated transmembrane protein, acts to detect cytoplasmic DNA species. STING homodimers were able to complex with self (apoptotic, necrotic) or pathogen related ssDNA and dsDNA and were indispensible for HSV-1-mediated transcriptional activation of a wide array of innate immune and pro-inflammatory genes in addition to type I IFN. Our data indicates that STING instigates cytoplasmic DNA-mediated cellular defense gene transcription and facilitates adoptive responses that are required for protection of the host. In contrast, chronic STING activation may manifest inflammatory responses and possibly autoimmune disease triggered by self-DNA.
α4β7 integrin-mediated lymphocyte adhesion to VCAM-1 under physiologic shear requires L-selectin function (P5116)
Authors: Loppnow Jessica , Kadono Takafumi , Sato Sinichi , Tedder Thomas , Steeber Douglas 
Published: January 2023

Abstract
Lymphocyte migration is facilitated by a cascade of adhesion molecule interactions. Specifically, lymphocyte rolling is mediated by selectins, while slow rolling and adhesion are attributed to integrin and Ig family members. α4β7 integrin interactions with its primary ligand, MAdCAM-1, synergize with L-selectin function to regulate the recirculation of naïve lymphocytes to the gut. However, subsets of regulatory T cells also express high levels of L-selectin and α4β7 integrin, and may instead bind to VCAM-1, a secondary ligand for α4β7 integrin. Therefore, potential synergism between L-selectin function and α4β7 integrin/VCAM-1 interactions were examined. To determine the ability of α4β7 integrin to interact with VCAM-1 under conditions of shear, transfected TK-1 T cell interactions with EA.hy926 endothelial cell lines were assessed using an in vitro flow chamber assay. While TK-1 cells, expressing α4β7 but not α4β1 integrin, showed little interaction with VCAM-1-transfected endothelial cells, the presence of L-selectin function enabled α4β7 integrin/VCAM-1 interactions to support slow rolling and adhesion. Importantly, α4β7 integrin interactions with VCAM-1 supported slow rolling and adhesion of murine regulatory T cells under shear stress in the presence of L-selectin function. Thus, functional synergy between L-selectin and α4β7 integrin/VCAM-1 interactions may promote regulatory T cell recruitment to sites of inflammation.
Isoforms of vitamin E differentially regulate inflammation and lung function (P5136)
Authors: Cook-Mills Joan , Abdala-Valencia Hiam , Berdnikovs Sergejs 
Published: January 2023

Abstract
Vitamin E regulation of disease has been extensively studied but most studies focus on the α-tocopherol isoform of vitamin E. These reports indicate contradictory outcomes for anti-inflammatory functions of the α-tocopherol isoform of vitamin E with regards to animal and clinical studies. The seemingly disparate results are consistent with our studies demonstrating that purified natural forms of vitamin E have opposing regulatory functions during inflammation. We have determined that α-tocopherol inhibits whereas γ-tocopherol elevates allergic inflammation, airway hyperresponsiveness, leukocyte transendothelial migration, and endothelial cell adhesion molecule signaling through protein kinase Cα. Moreover, we determined that α-tocopherol is an antagonist and γ-tocopherol is an agonist of PKCα through direct binding to a regulatory domain of PKCα. Furthermore, in a recent clinical study, we determined that increasing serum concentrations of γ-tocopherol associated with worse lung function, while increasing serum concentrations of α-tocopherol associated with better lung function. In summary, we have determined mechanisms for opposing regulatory functions of α-tocopherol and γ-tocopherol on inflammation. Information from our studies will have significant impact on the design of clinical studies on inflammation and on vitamin E consumption.
The divergent roles of Leptin and Wnt5a on mast cell function (P6020)
Authors: Kolawole Elizabeth , Ryan John 
Published: January 2023

Abstract
The prevalence of obesity has reached epidemic proportions in western countries. Increased levels of adipose tissue are strongly associated with disease states such as chronic heart disease, atherosclerosis, insulin resistance and type II diabetes. Obesity is also associated with increased risk of inflammatory conditions such as allergic asthma and allergic rhinitis. Furthermore, the obese state has been described as chronic low-grade inflammation. Thus, obesity and inflammation are inextricably linked. The role of mast cells in inflammatory disease and allergic reaction is unequivocal. Moreover, mast cells are instrumental effector cells in IgE mediated allergic diseases such as asthma and rhinitis. The receptor for IgE is the high affinity multimeric Fc receptor FcϵRI, which when activated through an antigen on mast cells results in signal transduction. This includes degranulation and the production of inflammatory and chemotactic mediators. We hypothesize that the adipokine Wnt5a enhances IgE mediated mast cell function and migration. We further hypothesize that the adipokine leptin suppresses IgE mediated mast cell cytokine production, a phenomenon seen in the Th1 prone genetic background (C57BL/6) but absent in the Th2 prone genetic background (BALB/c and 129/sv).
Mast cell-MDSC interactions promote chemokine secretion and parasite expulsion. (P6022)
Authors: Ryan John , Morales Johanna , Urban Joseph , Conrad Daniel 
Published: January 2023

Abstract
Mast cells and myeloid-derived suppressor cells (MDSC) are both increased at sites of growing tumors and parasitic infection. We have tested the effects of mast cell-MDSC interactions in vitro and in vivo. Surprisingly, co-culture of these cells resulted in a significant increase in pro-inflammatory chemokine production, which was attributed to Ly6C+ MDSC. MDSC depletion via gemcitabine treatment in mice infected with Nippostrongylus brasiliensis or Trichenlla spiralis drastically increased parasite burden, reminiscent of mast cell-deficient mice. Gemcitabine also decreased mast cell-dependent antigen-induced airway hyperresponsiveness (AHR) and inflammation, while MDSC injection worsened AHR. These data suggest that the evolutionary purpose of MDSC may be to potentiate Th2-induced, mast cell-dependent inflammatory responses.