bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-1 signaling directs the prioritization of inflammatory sites in a model of post-operative pulmonary infection
Authors: Crane Meredith , Xu Yun , Monaghan Sean F , Hall Benjamin M , Henry William L , Tran Holly L , Albina Jorge E , Jamieson Amanda 
Published: August 2023

Abstract
Individuals recovering from surgery are at risk of nosocomial infections such as pneumonia. In this setting, the innate immune system faces competing inflammation in the injured skin and infected lung, raising the question of how the immune response prioritizes inflammatory sites. We hypothesized that the initiation of a lung infection would disrupt wound healing. Supporting this hypothesis, data from the ACS NSQIP database showed that the rate of laparotomy dehiscence in surgical patients with pneumonia was over double the rate of those without pneumonia. We next developed mouse models of post-operative pulmonary infection to assess cellular mechanisms. Using the tail skin excision and dorsal subcutaneous PVA sponge implantation wound healing models, we found that bacterial lung infection slowed the rate of wound closure and impaired wound neutrophil, monocyte, and cytokine/chemokine responses. Lung infection quickly suppressed wound fluid IL-1b and downstream chemokine concentrations within 6 hours of inoculation. IL-1 receptor antagonist (IL-1RA), an inhibitor of IL-1 signaling, was upregulated in the bronchoalveolar lavage fluid and plasma 6 hours after lung infection. Administration of IL-1RA to the wounds of uninfected mice phenocopied the effects of lung infection. Conversely, rIL-1b rescued wound inflammation when administered to the to the wounds of mice with lung infection, although this occurred at the expense of lung bacterial clearance. These results suggest that the infected lung induces IL-1RA systemically as a protective mechanism to tune inflammation at distal sites via IL-1b suppression, allowing the innate immune system to prioritize inflammatory insults.
Supported by grants from Defense Advanced Research Projects Agency (DARPA), Brown University (Dean’s Areas of Emerging New Science Award, Carney Institute Innovation Awards), and NIH (NIES T32-ES7272 Training in Environmental Pathology; NIGMS COBRE Award P20GM10935, NHLBI R01 126887),
An invertebrate model using Galleria mellonella to study the innate immune response to Staphylococcus haemolyticus
Authors: Mares Chris Anthony , McGehee Rebecca , Smyth Davida 
Published: August 2023

Abstract
Staphylococcus haemolyticus is a normal inhabitant of healthy skin and is also an emerging cause of nosocomial infections in the immunocompromised. S. haemolyticus infections often lead to bacteremia, septicemia, peritonitis, endocarditis and various other outcomes. The goal of this project was to establish an invertebrate infection model for studying the innate immune response to Staphylococcus haemolyticus infections using the wax moth larva (Galleria mellonella). We have found that injecting wax worm larvae with 106 to 107 cfu/μL of S. haemolyticus consistently induced a rapid response and larval death within 24 hours at the higher dose range. We have characterized the general histopathological changes associated with infection and have started to characterize the general innate immune response as well as the hemocyte response to S. haemolyticus. Our findings support the usefulness of this model as an initial screening tool for systemic microbial infections.
Supported by grants from the Research Council and the College of Arts & Sciences at Texas A&M University-San Antonio.
Deciphering immunological mechanisms underlying the efficacy of nanoparticle-based vaccines against opioid use disorder (OUD)
Authors: Hamid Fatima Alamin Awad Alkareem , Schactler Scott , Walter Debra , Amin Hardik , Le Minh , Burkhart David , Zhang Chenming , Pravetoni Marco 
Published: August 2023

Abstract
The highly complex OUD and overdose epidemic poses a huge public health and economic burden, urging the need for safe and more effective therapeutic options. Vaccines offer a promising strategy for OUD treatment and prevention of overdose. Vaccinating against opioids generates drug-specific polyclonal antibodies (Ab) that selectively bind targeted opioids, blocking their distribution to the brain and thus blunting opioid induced rewards, pharmacological and side effects. Previous clinical trials of addiction vaccines showed proof of efficacy only in subjects with the highest Ab titers, highlighting the need to design more effective vaccines. The generation of a robust anti-drug polyclonal Abs response relies on CD4+ T cell-dependent B cell activation, hence dendritic cells (DC) are crucial for initiating CD4+ T cells priming and B cell responses. To improve vaccine formulation, we are exploiting a lipid-polymer hybrid nanoparticle (LPNP) platform and toll-like receptor (TLR) agonists to potentiate antigen recognition by antigen presenting cells (APCs) and subsequently the efficacy of anti-opioid conjugate vaccines. Using a flow cytometry-based in vitro activation assay, we demonstrated that LPNP-based vaccines are more effective in inducing macrophages activation marker (iNOS) and DCs co-stimulatory molecules and maturation markers (CD86, CD40, and MHC II) compared to conventional vaccines. Adding selected TLR agonists further improved nanovaccine efficacy in inducing the maturation and activation of APCs. These findings were confirmed with RT-qPCR quantifying mRNA expression of above-mentioned markers. These studies will identify key APCs subsets contributing to vaccine efficacy against OUD and other targets.
Supported by grants from NIH (UG3/UH3 DA048775)
Uncovering alterations in the murine interferon response due to dsRNA mycovirus during Aspergillus fumigatus infection
Authors: Rapp Alexander , Wang Xi , Ross Brandon , Tanner Christopher , Caffrey-Carr Alayna , Cramer Robert , Obar Joshua J , Olive Andrew , Thomas Sean 
Published: August 2023

Abstract
Aspergillus fumigatus is a mold that causes an array of diseases, the most severe form being invasive aspergillosis which occurs in highly immunocompromised individuals. We demonstrated that tissue-resident alveolar macrophages are critical for maintaining host resistance against Aspergillus fumigatus due to their role in the early type I/III interferon response. Specifically, CD11c-Cre x Mavsfl/fl mice have a decreased interferon response and survival outcomes when challenged with A. fumigatus, which can be functionally restored via adoptive transfer of wild-type SiglecF+ alveolar macrophages. Why alveolar macrophages are potent inducers of the type I/III interferon response after Aspergillus fumigatus challenge remains elusive. Here, we demonstrate that TGF-beta metabolically programs alveolar macrophages to be potent interferon inducers via enhanced dependency on oxidative phosphorylation and peroxisome biogenesis. The type I/III interferon responses were partially related to the inability of alveolar macrophages to recognize mycoviral double-stranded RNA (dsRNA). Aspergillus fumigatus strains infected with polymycovirus 1 (PmV1) are shown to induce a robust type I/III interferon responses both in vitro and in vivo. Critically, Mda5/MAVS-dependent interferon response can still be induced in a fungal dsRNA independent manner, which warrant further explorations. In this study we identified key metabolic traits of alveolar macrophages which enable robust Mda5/MAVS signaling necessary for initiating the Aspergillus fumigatus induced interferon response, which maintains host resistance against this ubiquitous mold, through both fungal dsRNA-dependent and -independent mechanisms.
Intravenous BCG Induction of Innate Memory in Young Pigs
Authors: Byrne Kristen A , Loving Crystal L 
Published: August 2023

Abstract
Exposure to vaccine strain of Mycobacterium bovis(M. bovis), Bacillus Calmette-Guerin (BCG), alters innate immune cells through epigenetic and metabolic modifications resulting in heightened responses to subsequent microbial insult (innate memory). As a biomedical model for human disease, pigs provide a critical space to study the role of innate memory in disease resistance. Young pigs were inoculated intravenous (IV) or intraperitoneal (IP) with either live or heat-inactivated BCG (IV-live, IP-live, IV-inactive, noBCG groups). At 2wks post BCG inoculation, monocyte production of IL-1β nor TNF cytokines following ex vivo stimulation with LPS was not different in any BCG group compared to noBCG, i.e. no innate memory. As dose of live BCG was lower than anticipated, at 3wks post primary inoculation, all pigs were inoculated a second time with respective preparations and routes of BCG. Ex vivo cytokine production in LPS stimulated monocytes was measured again at 5wks post primary inoculation (2wks post-secondary inoculation) and both IL-1β and TNF cytokines were elevated in IV-live (innate memory), but not IP-live or IV-inactive groups. Additionally, ex vivo stimulation of PBMC with homologous recall antigen (purified proteins from M. bovis) resulted in a 3-fold higher IFNγ response in IP-live compared to IV-live and no IFNγ production in noBCG or IV-inactive. Together these data suggest that while BCG can induce a state of innate memory in monocytes of young pigs, induction of training cannot be inferred by T cell production of IFNγ to recall protein antigen. Additionally, data continue to highlight pigs as a valuable model as multiple BCG factors (i.e. route, dose, or viability) may need to be considered for induction of innate memory.
USDA-ARS CRIS project 5030-32000-225-000D
SARS-CoV-2 and SARS-CoV-2 Spike protein S1 subunit Trigger Proinflammatory Response in Macrophages in the Absence of Productive Infection
Authors: Miller Lindsay Grace , Chiok Kim , Bose Santanu , Miura Tanya A 
Published: August 2023

Abstract
One of the hallmarks of critically ill COVID-19 patients infected with SARS-CoV-2 is exaggerated inflammatory response. Though macrophages mediate inflammatory responses and can produce pro-inflammatory cytokines to eliminate pathogens, infection with SARS-CoV-2 has been shown to cause immune dysfunction, leading to hyperinflammation in the lungs. To further understand the role of macrophages in hyperinflammatory responses during SARS-CoV-2 infection, we infected a THP-1 human derived macrophage cell line with SARS-CoV-2. Our results show that, though macrophages do not support viral replication, infection with SARS-CoV-2 still results in the upregulation of the mRNA of cytokines TNFα and CXCL10, which are markers of COVID-related hyperinflammation. In addition, we identified SARS-CoV-2 Spike protein S1 subunit as one viral factor involved in the upregulation of cytokines in macrophages. We show that glycosylated, soluble S1 protein can upregulate TNFα and CXCL10 mRNAs, as well as the secretion of TNFα, in macrophages in the absence of virus infection. Therefore, macrophage activation by the S1 subunit and SARS-CoV-2 infection may contribute to the hyperinflammation in the lungs seen in critically ill patients through the upregulation of proinflammatory cytokines such as TNFα and CXCL10.
Supported by NIH grant (R01 AI083387), Washington Research Foundation, and NIH NIGMS Predoctoral Biotechnology Training Grant 5T32GM008336
Ebola virus protein VP40 induces accessory cell-mediated activation of human natural killer (NK) cells
Authors: Singh Karnail , Le Hung 
Published: August 2023

Abstract
Natural killer (NK) cells have been implicated in Ebola virus disease (EVD) pathogenesis as well as in protection against Ebola viruses. A clear understanding of their role during EVD is needed for better exploiting their functionalities in designing effective countermeasures against Ebola viruses. Previously, we reported that Ebola virus protein VP40 induces IL-12 and IL-18 dependent activation of human NK cells that involves accessory cells. Here, we extended our studies to define the identities of the accessory cells and molecular mechanism(s) underlying their role in triggering NK cell activation. Purified CD14+ myeloid cells, CD3+ T cells or CD19+ B cells, isolated from normal human peripheral blood mononuclear cells (PBMCs), were co-cultured with purified CD56+ NK cells; cultures activated with VP40 and expression of IFN-γ, granzyme B, perforin, and surface CD107a in NK cells was measured. A small, but significant, increase in NK cell activation was observed when purified NK cells alone were incubated with VP40. Inclusion of CD14+ myeloid cells or CD3+ T cells in NK cell cultures resulted in significant additional increase in NK cell activation status. Both soluble factors and cell-cell contact dependent mechanisms were involved in VP40-induced, accessory cell dependent NK cell activation. In addition, a small subset of NK cells bound directly to VP40 and was triggered. Together, our data decipher both accessory cell dependent and independent mechanisms involved in Ebola VP40 induced NK cell activation, findings that may help in designing better anti-Ebola countermeasures.
Supported by funds from Cincinnati Children’s Research Foundation.
Neutrophilia and neutrophil-derived extracellular traps as innate immune markers of SARS-CoV-2 infection in a feline animal model
Authors: Gunasekara Sachithra J , Ludwig Breya , Cowan Shannon , Selvan Miruthula Tamil , Ritchey Jerry , Miller Craig , Rudd Jennifer 
Published: August 2023

Abstract
Excessive neutrophil recruitment and dysregulation contribute to worsened clinical outcomes in hospitalized COVID-19 patients. Targeting this dysregulated response offers therapeutic potential in the treatment of SARS-CoV-2 infections which can be enhanced by the availability of reliable, translational animal models for disease. Domestic cats are naturally infected and exhibit clinical disease similar to that seen in COVID-19. Utilizing our previously developed feline model for COVID-19, we hypothesize that neutrophil dysregulation and the release of neutrophil extracellular traps (NETs) significantly worsen pulmonary damage in domestic cats. Specific-pathogen-free cats (n=12) were inoculated with SARS CoV-2 (B.1.617.2) or vehicle (n=6). Plasma, bronchoalveolar lavage, and tissues were collected over 12 days. Detection of NETs using MPO-DNA ELISA and Quant-iT PicoGreen® dsDNA assay indicate significant elevations at 4dpi and 8dpi in the plasma as compared to vehicle, with the peak NETs detection at 4dpi(p&lt;0.05). These findings were similarly evident in the BAL fluid of infected versus vehicle-inoculated cats(p&lt;0.05). Immunohistochemistry of respiratory tissues utilizing three major markers of activated neutrophils and NETS; citrullinated histone 3, MPO, and neutrophil elastase, show marked amplifications compared to vehicle. These findings co-localize to areas of significant pulmonary damage. Elucidating these innate markers and their functional changes provides insight into understanding the immunopathogenesis and potential therapeutic interventions in SARS-CoV-2 infection. These data enforce the role of domestic cats as a flexible translational model to develop these interventions.
NIH-NIGMS (P20GM103648)
MrgprD-expressing nonpeptidergic neurons suppress mast cell responsiveness via glutamate release
Authors: Zhang Youran , Edwards Tara , Zhang Shiqun , Sumpter Tina L , Kaplan Daniel H 
Published: August 2023

Abstract
Cutaneous mast cells underly numerous skin inflammatory processes. Our study reveals that the ablation of a subset of sensory nonpeptidergic neurons expressing MrgprD increases the expression of a mast cell gene module, including the activating receptor, Mrgprb2, leading to increased mast cell reactivity to MrgprB2 ligands, and cutaneous inflammation in several disease models, such as irritant dermatitis and rosacea. MrgprD-expressing neurons releases glutamate to inhibit mast cell responsiveness. Reducing glutamate release from MrgprD-expressing neurons in the periphery resulted in a hyperresponsive state of mast cells. The antagonism of glutamate receptors on cultured peritoneal cavity mast cells (PCMCs) enhances MC degranulation and phenocopied the genetic state in the absence of MrgprD-expressing neurons. We subsequently identifies the kainate receptor GluR6/KA2 as the glutamate receptor subtype expressed on dermal MCs. Two non-specific agonists of GluR6, and the highly selective GluR6 agonist SYM2081 suppress the degranulation of PCMC in response to MrgprB2 activation but not FcɛRI crosslinking in a dose-dependent manner. In vivo, intradermal delivery of SYM2081 suppresses compound 48/80 induced-dermal MC activation, and attenuates croton oil-induced irritant dermatitis, but not IgE-dependent passive cutaneous anaphylaxis. Together, these data demonstrate that glutamate acting via the GluR6/KA2 receptor promotes mast cell hypo-responsiveness during homeostasis and suggests that selective agonism of this pathway has the potential to reduce mast cell activation in vivo, which would be of significant utility in the treatment of diseases where mast cells represent a key component of the pathogenic pathway.
Dysregulation of natural killer cells’ receptor expression and cytotoxic function by hydroxychloroquine
Authors: Elemam Noha M , Talaat Iman M , Maghazachi Azzam A 
Published: August 2023

Abstract

Background and Aim
Natural killer (NK) cells are crucial players of the innate immune system, especially in the fight against tumors and infections. Besides, NK cells are known to contribute in inflammatory processes associated with multiple diseases. The drug hydroxychloroquine (HCQ) has been repurposed to be used for the treatment of malaria, autoimmune diseases, and COVID-19 infection. However, its direct mechanism of action on NK cells remains unknown. In this study, we aimed to examine the effects of HCQ on NK cell activity and function.

Materials and Methods
The human NK cell line NK92 was cultured and treated with HCQ at 1, 5, and 10 μM for 24 and 48 hours. Then, NK92 cell viability, NKG2D and CD107a expression, as well as NK92 cell cytotoxicity were assessed. Also, IFN-γ production was measured by ELISA after HCQ treatment. Statistical analysis was performed where p&lt;0.05 was considered statistically significant.

Results
Upon treatment with different concentrations of HCQ for 48 hours, NK92 cell viability significantly decreased in a dose-dependent manner, with the lowest viability noted at 10 μM of HCQ. Additionally, NKG2D and CD107a expression were significantly reduced upon treatment with 5 and 10 μM of HCQ. Similarly, the cytotoxicity of NK92 cells against K562 cells was reduced at both 5:1 and 10:1, effector to target (E: T) cell ratios, when 10 μM HCQ was used. In contrast, IFN-γ levels were significantly increased upon the treatment with 1, 5, and 10 μM HCQ.

Conclusion
This study highlights the effect of HCQ on activating receptor expression, cytotoxicity, and IFN-γ production of NK cells. Despite being an immunomodulatory drug, more studies are needed to further understand HCQ’s effects on NK cells and perhaps other effector cells.

None
Infiltrative monocyte derived adenosine deaminase 2 is associated with COVID-19 related acute respiratory distress syndrome
Authors: Tiwari-Heckler Silpa , Vierbaum Lisa , Merle Uta , Jiang Zhenghui Gordon 
Published: August 2023

Abstract
Acute respiratory distress syndrome (ARDS) is a severe complication of COVID-19 characterized by immune dysregulation. The pathogenesis for this, particularly the involvement of infiltrative monocytes, is poorly understood. Extracellular ATP released during inflammation and hypoxia is rapidly scavenged into adenosine, which is further converted into inosine by adenosine deaminase (ADA). ADA2 is an extracellular isoform of ADA secreted by activated monocytes and expressed by infiltrated CD16+CXCR3+ monocytes.
Herein, we aim to study the impact of monocyte-derived ADA2 in the pathogenesis of COVID-19 associated ARDS. Critically ill COVID-19 patients were assigned to the ARDS group based on Berlin criteria, while the non-ARDS group included COVID-19 patients without any or with low-flow oxygen therapy.
We observed elevated ADA activity in the serum of ARDS patients, reflecting increased level of the secreted ADA2 protein, when compared to the non-ARDS group. ARDS patients exhibited high level of circulating IL-6, CXCL8 and CXCL10. Notably, correlation analysis revealed a significant association between circulating ADA activity and CXCL10, a chemokine for CXCR3+ monocytes. Post-mortem bulk RNAseq analysis of lung tissues showed increased expression of the ADA2 gene, CECR1, along with CXCL10 suggesting intrapulmonary accumulation of ADA2 positive macrophages. These observations were further supported by the analysis of single-nuclear and spatial transcriptomic atlas of lung tissues of severe COVID-19 patients, which demonstrated a strong expression of ADA2 in CD16+ monocytes.
In summary, our data suggest a potential role of infiltrative monocyte-derived ADA2 in modulating the immune response in COVID-19 related ARDS.
Supported by the German Center for Infection Research (TI 07.005 to STH)
TNFα-Mediated Damaging Inflammation During Aspergillus fumigatus Pulmonary Infection is Regulated by Type I Interferon Receptor 2 (IFNAR2)
Authors: Shepardson Kelly , Roemer Julia , Benson Evelyn , Hall Monica , Rynda-Apple Agnieszka 
Published: August 2023

Abstract
Invasive pulmonary aspergillosis (IPA), caused by the fungus Aspergillus fumigatus, generally occurs in patients with suppressed or non-functioning immune systems. The recent increase in the number of cases of influenza and SARS-Cov-2-infected patients acquiring IPA suggests that severe respiratory viral infections may create a suppressed lung immune environment that allows for fungal infections to occur. We recently found that differential type I interferon (IFN) signaling, a classic anti-viral host response, regulates susceptibility to IPA. Specifically, we found that absence of the type I IFN receptor 2 (IFNAR2) subunit of the heterodimeric IFNAR1/2 receptor, leads to increased damage and inflammation in response to A. fumigatus lung infection, while absence of IFNAR1 did not. Although the Ifnar2−/− mice had higher morbidity, we unexpectedly found that Ifnar2−/− mice killed more A. fumigatus by 24 hours post-infection compared to WT and Ifnar1−/− mice. However, this early clearance of A. fumigatus did not prevent invasive disease from developing in the Ifnar2−/− mice as infection progressed, suggesting that the early damage responses were creating a favorable environment for invasive growth. By altering the inflamed environment of the Ifnar2−/− mice early during infection, via neutralization of TNFα, we were able to reverse the damage and morbidity in these mice back to WT levels. Further, our results suggest that altered myeloid effector cell responses in Ifnar2−/− mice during A. fumigatus infection contribute to the damage that occurs. Together, our results expand our understanding of how anti-viral mechanisms, via IFNAR2, are creating a permissive environment for fungal infections to occur through regulation of damage.
Supported by grants from NIH/NIGMS (INBRE P20GM103474), NIH/NIAID (K22AI153671), AAI Careers in Immunology Fellowship, Parker B. Francis Fellowship