bluuebunny/crossref_metadata_2025_split huggingface dataset:
P2X3 is a Female-Dominant Activator of Mast Cells
Authors: Kee Sydney Ann , Haque Tamara , Dailey Jordan , Ryan John J 
Published: January 2023

Abstract
Asthma is characterized by increased mucus production, airway hyperresponsiveness and airway inflammation, all promoted by mast cell activation. There is a clear bias for greater incidence and acuity of asthma among females, but the reasons for this are not clear. Women are 60% more likely to be diagnosed with asthma and three times more likely to be hospitalized for it. In a search for possible new and upcoming drugs that might be repurposed to suppress mast cell function, we discovered that the ATP receptor P2X3, most often studied in neuropathic pain, has a role in mast cell activation. The P2X3 antagonist BLU-5937 suppressed IL-33 mediated-mast cell cytokine secretion in vitro and reduced neutrophilic peritonitis in vivo in female mice. In contrast, male mice and mast cells cultured from them showed no suppression when treated with BLU-5937. In addition, female mast cell responses were much stronger than males when activated with IL-33 in vitro. We also found that IL-33 elicits rapid ATP secretion by mast cells, possibly forming an autocrine loop via P2X3. These data coincided with little detectable P2X3 expression on male mast cells. We hypothesize that IL-33-mediated mast cell activation is more potent in females due to P2X3 receptor signaling. Our data support the theory that P2X3 is a female-dominant mast cell activator that can be targeted to reduce allergic inflammation. This research is novel because it could explain the sexual dimorphism in allergic disease and elucidate the role of P2X3 in the immune system.
Supported by 1RO1 AI164710
Identification of redundancy between human FcɛRIβ and MS4A6A proteins points toward additional mechanisms for FcɛRI trafficking and signaling
Authors: Bitting Katie Jane , Hedgespeth Barry , Ehrhardt-Humbert Lauren C , Arthur Greer , Bradding Peter , Tilley Stephen L , Cruse Glenn 
Published: January 2023

Abstract
Allergic diseases are triggered by signaling through the high affinity IgE receptor, FcɛRI. In mast cells (MCs) and basophils, FcɛRI is a tetrameric receptor complex comprising a ligand binding α subunit (FcɛRIα); a tetraspan β subunit (FcɛRIβ) responsible for trafficking the complex and amplifying signaling; and a signal transducing dimer of single transmembrane γ subunits (FcɛRIγ). However, FcɛRI also exist as presumed trimeric complexes that lack FcɛRIβ and are expressed on several cell types outside of the MC and basophil compartments. There are known differences in the trimeric FcɛRI complex between humans and mice and questions remain as to how this trimeric FcɛRI complex traffics and whether it signals in the absence of FcɛRIβ. We have previously reported that targeting FcɛRIβ with exon skipping oligonucleotides diminishes IgE-mediated degranulation in mouse MCs, but equivalent targeting in human MCs was not effective at reducing degranulation. Here, we report that an FcɛRIβ-like protein exists in humans and we identify that the Membrane Spanning 4A (MS4A) gene cluster member, MS4A6A compensates for FcɛRIβ in FcɛRI trafficking and signaling. The function of MS4A6A is currently unknown, but MS4A6A and FcɛRIβ are encoded by highly related genes within the same family (MS4A gene family) that cluster within the human gene loci 11q12–q13, a region linked to allergy and asthma susceptibility. Human MS4A6A promotes surface expression of FcɛRI complexes and promotes degranulation. Indeed, our data suggest the presence of either FcɛRIβ or MS4A6A is sufficient for degranulation. This study suggests that MS4A6A could be an elusive FcɛRIβ-like protein in human MCs that performs compensatory functions to FcɛRIβ.
Research was supported by the National Institutes of Health (NIH), National Institute of Allergy and Infectious Diseases (NIAID), Award Number R01AI143985, Department of Molecular Biomedical Sciences, College of Veterinary Medicine, start-up funds, Institutional National Research Service Award from the Office of the Director of National Institutes of Health, NIH T32OD011130.
RSV inhibits myeloid cell recruitment during S. aureus coinfection
Authors: Rich Helen Eva Adams , Morris Susan H , Lukacs Nicholas W , Moore Bethany B 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) infects almost all humans by the age of 2. RSV more often causes severe lower respiratory tract disease in younger children and older adults, which may be exacerbated by increased susceptibility to secondary bacterial infection (coinfection). Like influenza, it is thought that RSV may predispose the lung to opportunistic infections with strains of bacteria normally present as colonizers of the nasopharynx, predominantly Staphylococcus aureus and Streptococcus pneumoniae. While there has been significant investigation into mechanisms of RSV/bacterial coinfection, models that faithfully replicate the pathology of human RSV infection have not been fully explored. To examine this aspect, we infected mice with a strain of RSV (rA2-line 19F, 2×105 PFU) that induces significant IL-13, mucus, and airway hyperresponsiveness. We found that RSV infection six days prior to S. aureus infection (5×107 CFU) reduces bacterial clearance. Flow cytometry data suggest that RSV infection alters cellular recruitment in response to S. aureus, largely by reducing neutrophils and monocytes. Surprisingly, we saw increased levels of the monocyte chemokine CCL2, suggesting compensatory chemokine production in response to dampened cellular recruitment. Additionally, we observed no change in several cytokines that exacerbate inflammation during influenza/S. aureus coinfection, including IL-1β, IL-6, and TNFα. Together, these data suggest a markedly different mechanism of increased susceptibility to bacterial infection during RSV, characterized by reduced--rather than overexuberant--myeloid cell recruitment.
Supported by grants from NIH (R35HL144481 to Beth Moore, T32HL007517 to Helen Rich) and the Michigan Postdoctoral Pioneer Program at the University of Michigan Medical School.
Baseline NK cell frequencies are greater in individuals with asymptomatic SARS-CoV-2 infection compared to those that develop symptomatic illness
Authors: Graydon Elizabeth , Lindrose Alyssa , Anderson Stephen , Malloy Allison , Lizewski Stephen , Lizewski Rhonda , Weir Dawn , Goforth Carl , Sun Peifang , Letizia Andrew , Mitre Edward 
Published: January 2023

Abstract
As NK cells are critical for control of viral infections, we tested the hypothesis that individuals with asymptomatic SARS-CoV-2 infection have greater NK cell frequencies prior to infection than individuals with mild, symptomatic infection. Marine recruits who elected to participate in the COVID-19 Health Action Response for Marines (CHARM) study were prospectively followed for two months twice weekly using PCR testing for SARS-CoV-2 infection and symptom questionnaires. Peripheral blood mononuclear cells obtained prior to infection during enrollment in the study and at various time points post-infection from 47 participants diagnosed with asymptomatic infection and 49 with symptomatic infection were analyzed by flow cytometry for frequencies of total NK cells, NK cell subsets, and surface expression of activating and inhibitory receptors. Consistent with our hypothesis, frequencies of total (CD3− CD14− CD19− CD56+) NK cells were significantly greater at baseline in asymptomatic compared to symptomatic participants (10.4% vs 8.3%, p = 0.015). This difference was due to increased frequencies of mature (CD56dim CD16+) NK cells in asymptomatic individuals (76.3% vs 69.8%, p = 0.049). No differences were observed in baseline frequencies of immature (CD56bright CD16−) or adaptive (CD56dim CD16+ NKG2C+ CD57+) NK cells, or in surface expression of activating (NKG2C, NKG2D) or inhibitory (NKG2A, KIR2DL1, KIR2Dl2/L3/S2, KIR3DL1) receptors. Frequencies of total and mature NK cells decreased significantly post-infection, reaching a nadir at 3–4 weeks. In contrast, frequencies of immature NK cells increased by 4 weeks. These results suggest that NK cells may play a protective role against symptomatic SARS-CoV-2 infection.
This work was supported by a grant from the Defense Health Agency through the Naval Medical Research Center (work unit number 9700130).
Human β-defensin 2 (hBD-2) improves neutrophilia and bacterial clearance during cigarette smoke exposure
Authors: Milad Nadia , Pineault Marie , Beaulieu Marie-Josée , Aubin Sophie , Jensen Benjamin A H , Morissette Mathieu 
Published: January 2023

Abstract

Rationale
Treatment of cigarette smoke-associated lung diseases has largely focused on broad-spectrum anti-inflammatory therapies, such as inhaled corticosteroids. However, these therapies have recently been shown to enhance patient susceptibility to pneumonia.

Objective
Assess whether the anti-microbial peptide, human β-defensin 2 (hBD-2), can simultaneously reduce pulmonary inflammation in cigarette smoke-exposed mice while maintaining immune competence during bacterial exacerbation.

Methods
Mice were exposed to cigarette smoke acutely (4 days) or chronically (5 days/week for 7 weeks) and administered hBD-2 intranasally or by gavage. In a separate model of acute exacerbation, chronically exposed mice treated with hBD-2 were infected with non-typeable Haemophilus influenzae prior to sacrifice.

Results
In the acute exposure model, cigarette smoke-associated pulmonary neutrophilia was significantly blunted by both local and systemic hBD-2 administration. Similarly, chronically exposed mice administered hBD-2 therapeutically exhibited reduced pulmonary neutrophil infiltration and downregulated pro-inflammatory signaling in the lungs compared to vehicle-treated mice, as assessed via microarray. Finally, in a model of bacterial exacerbation, hBD-2 administration effectively limited neutrophil infiltration in the lungs while markedly reducing pulmonary bacterial load.

Conclusions
This study shows that hBD-2 treatment can significantly attenuate lung neutrophilia induced by cigarette smoke exposure while preserving immune competence and promoting an appropriate host-defense response to bacterial stimuli.
Supported by CIHR
Increased M1 macrophage activity makes females more sensitive to Francisella tularensis infection
Authors: Spencer Charles T , Sanchez Guillen Michelle A 
Published: January 2023

Abstract
The intensity of the inflammatory response differs between males and females, with females having a stronger basal inflammatory response compared with males. This leads to differences in sensitivity to autoimmune and infectious diseases. Males tend to be more susceptible to infectious diseases whereas females tend to be more susceptible to autoimmune diseases. However, certain infectious diseases cause disease by over stimulating an inflammatory response resulting in dysregulation of cytokine production and generation of a cytokine storm. We hypothesized that the elevated basal inflammatory state of females would make them more susceptible to inflammatory infectious diseases such as Francisella tularensis. Indeed, survival and immunological studies demonstrated that females had a stronger innate immune response and were more likely to succumb to tularemic-disease compared with males. Importantly, this dichotomy was independent of hormonal signaling. We traced the source of this elevated inflammatory response to alterations in the M1 and M2 macrophage activation profile. In females, M1 macrophages were activated earlier than in males in response to inoculation. These M1 macrophages from female mice showed a stronger activation both in vivo and in vitro cultures. This was consistent in both mouse and human systems. Our results demonstrate that females possess a higher abundance of M1 macrophages and that these M1 macrophages are more inflammatory in response to F. tularensis infection. This suggests that the consequence of the dichotomy between the male and female inflammatory response is dependent upon the mechanism of disease triggered by infectious diseases.
Supported by grants from the NIH (SC1 GM122699-01A1) and NSF (MRI 1626587).
To Sir with Love! Students Feedback from an Immunology Course.
Authors: Eshetie Kirubel , Ngo Sylvie , Hailu Hailu , Desta Negatu , Fazal Nadeem 
Published: January 2023

Abstract
Students are obligated to write reviews and evaluate both their courses as well as their teachers at the end of the semester in anonymous surveys. The response rates from students have been continuously on decline as reported by third party evaluation companies. This has resulted in both failure of the purpose of evaluations as well as validity and statistical significance of these responses. We provided a non-binding, open and forum-like tool to get the perceptions and trends from Chicago State University College of Pharmacy students. Anonymous survey was sent using google form at the end of an Immunology course being offered to year-2 pharmacy students. Data from 29 respondents were collected and most significant results are summarized. When asked about their learning style; 45% loved traditional PowerPoint presentations as compared to 7% recorded lectures, 10% made their own notes, 3% were small group learners, while 31% solely relied on studying practice tests. When asked about the use of YouTube; 62% preferred random searches while 39% found Khan Academy videos as very useful. When asked about major barrier in learning Immunology; 38% reported test anxiety, 14% personal issues, 10% overwhelmed with curriculum, and 7% cited lack of motivation. In response to assessment questions, 92% students suggested that Instructors should not have more than 60 questions in 2-hour exam. Regarding exam question types, 90% preferred MCQs, 7% Essay, 14% Case-based, and 31% True/False. Following a 16-week, 2-credit course of Immunology, 54% got motivated to do research in Immunology. When asked about overall sentiments after taking a first course in Immunology. The unanimous response was that they all felt as if they were taking a course in a foreign language.
Antibody ‘Hotspots’ induce antibody-dependent cell-mediated cytotoxicity against SARS-CoV-2 spike-expressing lung fibroblasts
Authors: Holder Kayla , Hatfield Keeley , Ings Danielle , Fifield Katie , Harnum Debbie , Grant Michael 
Published: January 2023

Abstract
Since COVID-19 emerged in 2019, neutralizing antibody (Ab) responses have dominated as surrogate measures of protection against infection and severe illness. The mostly self-limiting course of SARS-CoV-2 infection suggests the innate immune system, including natural killer (NK) cells, plays a role in its control. Therefore, we investigated SARS-CoV-2-specific antibody-dependent cell-mediated cytotoxicity (ADCC) and if Abs against Wuhan and early SARS-CoV-2 variants activate ADCC against emergent highly transmissible variants. ADCC was evaluated by measuring NK cell cytotoxicity (51Cr) against plasma (1:1000)-sensitized MRC-5 cells stably transduced to express Wuhan or Delta spike. ADCC mediated by plasma from double-vaccinated and convalescent adults was generally low (range 1.2 – 24.9%; median 7.2% (IQR 5.2 – 10.6%), and plasma from those with previous natural infection mediated significant ADCC (&gt; 10% lysis) to the same extent as plasma from fully vaccinated individuals (13/34 vs 11/44). Hybrid immunity imparted robust ADCC (27.9 ± 10.8% target lysis) in 32/34 individuals tested and levels of ADCC significantly correlated with IgG3 responses (P = 0.0093) against spike stalk (S2) domain. A peptide scan identified four distinct ‘hotspot’ regions associated with robust ADCC in individuals with reactive Abs. Overall, a minor decline in ADCC against Delta variant spike-expressing relative to Wuhan targets occurred in all groups; however, individuals with Abs against the region containing D614G mediated greater ADCC against Delta variant than Wuhan targets. Determining which Abs provide robust and broad ADCC can inform future COVID-19 prevention and treatment strategies and predict ADCC responses for emergent variants.
Supported by grants from CIHR through the COVID Immunity Task Force (VR1-173202)
Neutrophil NOX2 is Required to Control Experimental Urinary Tract Infection
Authors: Cotzomi-Ortega Israel , Han Rachel , Cortado Hanna , Li Birong , Becknell Brian , de Dios Ruiz-Rosado Juan 
Published: January 2023

Abstract
Urinary tract infections (UTIs) affect 150 million people worldwide. Uropathogenic Escherichia coli (UPEC) is the primary pathogen causing over 80% of UTIs. Systemic antibiotic treatment is the only available therapy for UTIs. Thus, novel non-antibiotic treatments for UTI are necessary. The NADPH oxidase complex 2 (NOX2) has an essential role in the antimicrobial response against many bacterial and fungal infections, but its role during UTIs remains unknown.

Objective
To study the role of NOX2 in bacterial killing during experimental UTI.

Methods
Female wild-type (WT) C57BL/6 mice, NOX2 knockout (KO) mice, and WT mice treated with a NOX2 inhibitor were transurethrally infected with UPEC (CFT073 strain). NOX2 protein expression was analyzed by Western blotting. Cellular sources of NOX2 were identified by immunofluorescence. Neutrophils were isolated from murine bone marrow. Human neutrophils were purified from peripheral blood of healthy donors. Purified neutrophils were infected with UPEC in vitro. UPEC burden was assessed by enumerating colony forming units (CFU). Reactive oxygen species (ROS) production was quantified using the Amplex™ Red kit.

Results
NOX2 KO mice, or mice treated with NOX2 inhibitor, were more susceptible to experimental UTI compared to WT mice. Immunofluorescence microscopy localized NOX2 primarily in infiltrating neutrophils. Neutrophils from NOX2 KO mice exhibited impaired extracellular ROS production and increased intracellular bacteria compared to WT controls, after challenged with UPEC in vitro. Similar effects occurred in human and murine neutrophils following chemical inhibition of NOX2 in vitro.

Conclusion
Neutrophil NOX2 serves a critical antimicrobial role during experimental UTI.
Supported by grant from NIH (K01 DK128379-01)
A novel function for the CoQ10 biosynthetic complex in anti-pneumococcal macrophage function
Authors: Walker Emma C , Todd Elizabeth , Ramani Rashmi , Anaya Edgar , Javati Sarah , Matlam John-Paul , Pomat William , Morley S Celeste 
Published: January 2023

Abstract
Despite the recent introduction of pneumococcal polysaccharide and conjugate vaccines, Streptococcus pneumoniae infection remains a leading cause of illness and death worldwide. In particular, infants in Papua New Guinea are at increased risk of severe pneumococcal pneumonia compared to infants in similar countries. We sought to determine if a novel genetic variant could explain this increased susceptibility. Whole exome sequencing revealed a single nucleotide variant (D308Y) in the gene encoding COQ6 (COQ6DY), a monooxygenase required for CoQ10 biosynthesis. We have utilized both a Saccharomyces cerevisiae model and a mouse model of COQ6DY to show that despite adequate production of CoQ10, this variant directly causes increased susceptibility to S. pneumoniae. This variant represents a previously unknown function of the CoQ10 biosynthetic complex that does not exert its effects through CoQ10 deficiency but rather through alterations of mitochondrial function and metabolism. These mitochondrial deficits in COQ6DY macrophages are sufficient to abrogate macrophage killing of S. pneumoniae and alter the coordination of the downstream immune response. In conclusion, we have identified a novel susceptibility allele to S. pneumoniae infection that exerts its effects via alterations in macrophage mitochondrial function.
Supported by NIH (R21 AI142723), SLCH Children’s Discovery Institute (PD-II-2018-742)
Examining the immune regulatory function of NLRP12 through novel protein interactions
Authors: Rippe Catherine A 
Published: January 2023

Abstract
NOD-like receptors (NLRs) are intracellular proteins that play an important role in the regulation of the innate immune response to pathogens. Since being identified, various functions for NLRP12 have been suggested. It has been shown to negatively regulate the inflammatory response by inhibition of the NF-kB signaling pathway and to exacerbate inflammation through the formation of a multi-protein complex called an inflammasome. Thus, the mechanisms by which this protein functions largely remain unclear. In this study, we examined the interaction between Cul3, a protein involved in protein ubiquitination, and NLRP12 to elucidate possible mechanisms by which it functions and/or is activated.
Supported by a grant from the Missouri State University Graduate College
Neutrophilic inflammation reprograms the bone marrow to impair T cell immunity during Tuberculosis
Authors: Saqib Mohd , Das Shreya , Mishra Bibhuti B 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb) infection induces persistent influx of neutrophils that precludes both CD4 and CD8 T cells from tuberculosis (TB) lesions in genetically susceptible mice. Depletion of neutrophils restores T cell number and ameliorated disease. High neutrophil to T lymphocyte ratio in the peripheral blood of pulmonary TB (PTB) patients correlate with sputum smear positivity and disease severity. These observations led us to hypothesize that infiltration of neutrophils negatively affect the normally protective T cell response in TB lesions. By employing various mouse strains that present neutrophil-rich lung lesions, we demonstrated that instead of serving as canonical myeloid derived suppressor cells (MDSCs), neutrophils produce granulocyte colony stimulating factor (CSF3/G-CSF) to indirectly regulate T cell numbers in TB lesions. Neutrophil-derived G-CSF reprograms the bone marrow towards a granulocyte-biased hematopoiesis at the expense of lymphocyte progenitors. Transcriptome profiling of the isolated neutrophils from the lung, spleen and bone marrow revealed a type I interferon (IFN-I) response signature in these neutrophils. Hematopoietic cell-intrinsic IFN-I signaling (IFNAR1) is both necessary and sufficient to promote the pathologic granulopoiesis generating Mtb-permissive neutrophils while suppressing lymphocyte progenitors (CLPs) production. Finally, blockade of IFNAR1-signaling ameliorates disease pathology by preserving the hematopoietic equilibrium. Collectively, our work uncovers a mechanism by which type I IFN response in neutrophils along with G-CSF perturbs hematopoiesis in the bone marrow and negatively impacts T cell immunity in TB lesions.