bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cessation from smoking improves innate host defense and clearance of experimentally inoculated nasal S. aureus
Authors: Cole Alexander M. , Schmidt-Owens Mary , Beavis Ashley C. , Chong Christine , Tarwater Patrick M. , Schaus James , Deichen Michael G. , Cole Amy Liese 
Published: January 2023

Abstract
Staphylococcus aureus (SA) nasal carriage is transient in most humans and usually benign, but dissemination of SA to extranasal sites causes the majority of clinical infections, and SA is a major cause of serious infections in the U.S. A better understanding of innate nasal decolonization mechanisms is urgently needed, as are relevant models for studying SA clearance. Here, we screened a population of healthy smokers for nasal SA carriage, and compared participants’ abilities to clear experimentally applied nasal SA before and after completion of a smoking cessation program. We determined that cigarette smoking increases mean nasal SA load (2.6×104 CFU/swab) compared to healthy non-smokers (1.7×103 CFU/swab) and might increase the rate of SA nasal carriage in otherwise healthy adults: 22 of 99 smokers carried SA at the screening visit, while only 4 of 30 non-smokers screened positive during the same time period. Only 6 of 19 experimental inoculation studies in active smokers resulted in SA clearance within the month of follow-up, while in the cessation group, 6 of 9 subjects cleared nasal SA and carriage duration averaged 21±4 days. Smoking cessation associated with enhanced expression of SA-associated IL-1β and G-CSF in nasal fluids. Participants who failed to clear SA exhibited higher nasal SA load and elevated nasal interleukin-1 receptor antagonist (IL-1RA) expression at the pre-experiment study visits. We conclude that smokers exhibit higher SA load than non-smokers, and that innate immune pathways including G-CSF expression and signaling through the IL-1 axis are important mediators of nasal SA clearance.
Purine nucleotide metabolism regulates expression of human MICA.
Authors: O’Callaghan Chris , McCarthy Michael T. , Moncayo Gerald , Hiron Thomas K. , Jakobsen Niels A. , Valli Alessandro , Soga Tomoyoshi , Adam Julie 
Published: January 2023

Abstract
Human MICA is upregulated in abnormal or stressed cells, including cancer cells, virus-infected cells and rapidly proliferating cells. Recognition of MICA by the activating NKG2D receptor on cytotoxic immune cells provides a mechanism for identification and potential elimination of aberrant cells. Despite the wide range of possible therapeutic applications of MICA manipulation, the factors that control MICA expression are unclear. We have used metabolic interventions and metabolomic analyses to show that the transition from quiescent cellular metabolism to a ‘Warburg’ or biosynthetic metabolic state induces MICA expression. Glucose transport into the cell and active glycolytic metabolism are both necessary to upregulate MICA expression. Active purine synthesis is required to support the effect of glucose on MICA expression and increases in purine nucleotide levels are themselves sufficient to induce MICA expression. This metabolic induction of MICA expression influences NKG2D-dependent cytotoxicity by immune cells. These findings are consistent with a model of MICA regulation whereby the purine metabolic activity of a cell can be indicated at the cell surface by MICA expression and is the subject of surveillance by NKG2D receptor-expressing immune cells.
The engagement of the aryl hydrocarbon receptor by tryptophan derivatives can prevent the development of anti-FVIII antibodies in an experimental model of hemophilia A
Authors: Matino Davide , Gargaro Marco , Scalisi Giulia , Manni Giorgia , De Luca Antonella , Puccetti Paolo , Quintana Francisco Javier , Alfonso Iorio , Fallarino Francesca 
Published: January 2023

Abstract
Patients affected by haemophilia A require treatment with factor VIII (FVIII) protein. The most relevant complication is the development of neutralizing FVIII-specific antibodies or “inihibitors”. We reported that the inhibitor-positive status was associated with reduced activity of the immune-regulatory enzyme indoleamine 2,3-dioxygenase 1 (IDO1), that promotes regulatory effects via the production of tryptophan catabolites, known as kynurenines. Some of those tryptophan derivatives are endogenous ligands for the Aryl hydrocarbon receptor (AhR). In this study we tested the potential of tryptophan-related AhR ligands for inhibiting the development of anti-FVIII antibodies in hemophilic (F8 KO) mice. To this aim, F8 KO mice were treated with recombinant human FVIII (rhFVIII) alone or in combination with selected AhR ligands once weekly for four weeks. Antibody titers were tested by specific ELISA and Bethesda test. All mice treated with rhFVIII developed high-titer anti-FVIII antibodies after 4 weeks of treatment. Administration of a specific tryptophan metabolite prevented the generation of anti-FVIII antibodies in almost 80% of F8 KO mice. The protective effect of these AhR ligands was negated by co-administration of the AhR antagonist CH-223191 or in AhR KO mice. Similar results were obtained by administration of engineered gold nanoparticles loaded with the same tryptophan metabolite and rhFVIII. These results suggest that the engagement of AhR, by specific tryptophan derivatives, may be a possible new strategy to control the immune response to rhFVIII. Our findings might lead to the development of novel immunomodulatory interventions for preventing or eradicating inhibitors in hemophilia A patients.
Regulation of NKT cell metabolism by PLZF
Authors: Kumar Ajay , Pyaram Kalyani , Yarosz Emily , Giri Shailendra , Chang Cheong-Hee 
Published: January 2023

Abstract
Cellular metabolism and signaling pathways are the key regulators to determine T cell fate, survival and function, particularly during activation. Resting CD4 and CD8 T cells use oxidative phosphorylation as a primary energy source but switches to glycolysis upon activation, which is necessary to produce biomolecules for cell proliferation and function. Failure of this reprogramming is detrimental for T cell mediated immunity. However, little is understood about how NKT cells control their metabolism to survive and function. We found that NKT cells operate distinctly different metabolic programming from CD4 T cells for their survival and function. Supporting evidence includes that, unlike CD4 T cells, oxidative phosphorylation seems to be preferred over glycolysis in NKT cells even after stimulation as revealed by lower level of intracellular lactate produced by NKT cells than CD4 cells after activation. These cells also seem to have high energy as revealed by higher ATP in NKT cells. Although the proper maintenance and functions of stimulated NKT cells need glucose, they are sensitive to the elevated glycolytic potential compared to CD4 T cells resulting in spontaneous cell death accompanied by hyperproliferation. Importantly, PLZF is the essential regulator of NKT cells’ metabolism so that similar changes of glucose metabolism were found in CD4 T cells expressing PLZF. Conversely, NKT cells with haplodeficient PLZF were more tolerant to increased glycolysis, strongly supporting the key role of PLZF in NKT cell metabolism and homeostasis. We are currently investigating the underlying mechanisms by which PLZF controls metabolism in NKT cells using several approaches including metabolomics.
Pseudomonas aeruginosa-induced lung epithelial production of IL-8 contributes to PMN sub-mucosa recruitment but not trans-epithelial migration in vitro.
Authors: Lanter Bernard B. , Eaton Alex D. , Gipson Kevin S. , Yonker Lael M. , Hurley Bryan P. 
Published: January 2023

Abstract

Background
Numerous lung associated diseased states such as acute pneumonia and cystic fibrosis (CF) are associated with increased concentrations of IL8 and infiltration of neutrophils (PMNs) in the airspace. IL8 production by lung epithelial cells (ECs) is stimulated by Pseudomonas aeruginosa infection, an opportunistic pathogen involved in acute pneumonia, and lung infection of CF patients. The exact role IL8 plays in migration of circulating PMNs into the infected lung and airspace is still uncertain.

Objective
Assess the role of IL8 in mediating PMN migration in response to P. aeruginosa infected lung epithelium.

Methods
CRISPR/Cas9 was used to delete the IL8 gene from human lung ECs, which were grown as polarized monolayers in a co-culture transwell model. Basolateral to apical PMN migration to PAO1 infection was assessed. A novel double transwell model was developed to observe PMN submucosa migration in response to PAO1 infection in the presence and absence of IL8 lung EC production.

Results
PAO1 induced PMN trans-epithelial (TE) migration was not impacted in the absence of IL8 expression. When PMNs were placed in a separate transwell suspended 5 mm above the basolateral side of the infected epithelium, the IL8 gradient produced was sufficient to cause relevant migration of PMNs to the basolateral side of the infected epithelium. The level of migrated PMNs in the double transwell system with tissues devoid of the IL8 gene was equivalent to the presence of PMNs which responded to un-infected epithelial tissues.

Conclusion
These findings further support that IL8 is integral in PMN recruitment, and suggests that IL8 is most important towards recruiting PMNs to the submucosa during P. aeruginosa PAO1 infection of lung epithelium.
Ablation of adaptive immune responses facilitates increased bacterial clearance in murine model of defective mucociliary clearance
Authors: Lewis Brandon W. , Patial Sonika , Saini Yogesh 
Published: January 2023

Abstract
Defective mucociliary clearance (MCC) associated with muco-obstructive lung diseases may compromise effective clearance of aspirated bacteria. Mucus hypersecretion is often associated with mucous cell metaplasia (MCM) of airway epithelium, which is a Type 2 immune response. Whether ablation of Type 2 response abolishes MCM/muco-obstruction, and thus, enhances bacterial clearance remains untested. Accordingly, we hypothesized that the ablation of adaptive immune responses will promote bacterial clearance in murine model of defective MCC, i.e., Scnn1b [Sodium channel, non-voltage-gated 1, beta] transgenic (Tg+) mice. The Tg+ model recapitulates hallmarks of muco-obstructive airways, including MCM and bacterial infections. The Type 2 response deficient (Il2rg−/−, Rag1−/− and Il2rg −/−\Rag1−/−) mice strains on Tg+ background were observed at neonatal age (6–7 days) for bacterial clearance response. Our data suggest that while Il2rg−/− mice [lacking innate lymphoid cells (ILC)] had significantly impaired bacterial clearance in control (Tg−) as well as Tg+, the Rag1−/−mice [lacking adaptive immune response] did not exhibit impaired bacterial clearance in Tg− as well as Tg+ neonates. The defective bacterial clearance in Il2rg−/− was not due to reduced phagocyte recruitment. Interestingly, Il2rg −/−\Rag1−/− mice had significantly improved bacterial clearance, an observation that remains a focus of our ongoing investigation. Our analyses revealed absence of IL-10 in Rag1−/−mice was associated with morphological activation of macrophages. Taken together, our current data suggest that while ILCs are essential for bacterial clearance, the adaptive immune responses suppress macrophage’s antibacterial function.
Early, in situ re-activation of lung memory T cells drives chronic lung inflammation
Authors: Turner Damian L. , Tanna Nicole , Barahona Josselyn 
Published: January 2023

Abstract
Acute immune responses within the respiratory tract generate memory T lymphocyte populations that are retained within the lung tissue and contribute to chronic inflammatory responses upon subsequent re-exposures to antigen. The generation and maintenance of antigen-specific memory T cell populations within the lung are therefore processes which, if inhibited, could ameliorate the symptoms of chronic inflammatory lung diseases. We sought to develop a model system that consistently generates lung inflammation and antigen-specific memory T cells within the lung. We show that prolonged respiratory exposure to lipopolysaccharide and a protein antigen generates a long-lived memory T cell population in the lung. These cells are retained in this compartment even after a week-long period of induced lymphopenia. Furthermore, lung memory T cells are re-activated in an antigen-specific manner upon intranasal challenge with antigen. The early in situ reactivation of memory T cells in the lung induced the recruitment of innate immune cells into the lung parenchyma and into the airways. Thus, elucidating the pathways involved in the tissue retention of the lung-retained memory T cell population may lead to therapies that can reduce harmful T cell populations within the lung.
A novel function of CCL27 in the localization of skin-homing CCR10+ lymphocytes
Authors: Davila Micha L. , Xiong Na 
Published: January 2023

Abstract
The skin contains an abundance of resident lymphocytes that play vital roles in the first line of defense. Dysregulation of the skin immune system could lead to diseases such as psoriasis and atopic dermatitis; studying the immune cells that contribute to skin immune homeostasis is crucial to understanding these diseases. Our lab has found that various subsets of skin-homing lymphocytes that express the chemokine receptor CCR10 have homeostatic functions. The skin specific ligand to CCR10 is CCL27. We found that the expression pattern of CCL27 in the skin of mice is similar to humans: low expression at a young age with a gradual upregulation into adulthood. This upregulation of CCL27 does not require the stimulation of skin colonizing bacteria, as germ free mice did not significantly differ from specific pathogen free mice in their expression of CCL27, suggesting an intrinsic regulatory mechanism for CCL27 expression. CCR10-expressing cells were found to concentrate near the hair follicles, suggesting a role for CCL27/CCR10 in the localization of these cells to specific microstructures. Preliminary analysis of skin-draining lymph node sections from CCR10+/EGFP vs CCR10EGFP/EGFP mice show differential localization of lymphocytes around the vessel-rich medulla. Further, lymphatic endothelial cells from the skin draining lymph nodes express CCL27. These findings indicate a novel role of CCL27 in directing the migration and localization of CCR10+ cells within the skin-draining lymph nodes and skin.
T Cell-Directed IL-17 production by Lung Granular γδ T cells is Coordinated by a Novel IL-2 and IL-1β Circuit
Authors: Menoret Antoine , Buturla James A. , Xu Maria Mei , Svedova Julia , Kumar Sanjeev , Rathinam Vijay A. , Vella Anthony T. 
Published: January 2023

Abstract
Immune-mediated lung injury is considered the result of an exacerbated innate immune response, although a role for adaptive lymphocytes is emerging. αβ T cells specific for S. aureus enterotoxin A orchestrates a typical innate-like Tγδ17 response during lung injury. However, the mechanism by which a specific T cell response drives an innate immune response in the lung is unclear. Here, we show a role for IL-2 triggering IL-17 production by lung granular γδ T cells as IL-17 synthesis was reduced by IL-2 blocking mAbs in vitro and in vivo. Granular γδ T cells in the lung constitute a rare population, hence Mass Cytometry (CyTOF) was utilized to characterize this population. CyTOF analysis revealed that lung γδ T cells responded directly to IL-2 as evident from STAT5 phosphorylation and RoRγt expression. IL-2 receptor blocking mAbs and JAK inhibition impaired STAT5 phosphorylation and IL-17 release. Moreover, inhalation of S. aureus enterotoxin A induced IL-2 secretion and caspase-1-dependent IL-1β activation to drive IL-17 production. This T cell-mediated inflammasome dependent IL-17 response is maximum when lung Tγδ17 cells were sequentially stimulated first with IL-2 then IL-1β. Further investigation is still needed to identify the cellular source of IL-1β. Interestingly, when IL-2 is given therapeutically to cancer patients it carries a known risk of lung injury that is largely indistinguishable from that seen in sepsis. Hence, this novel mechanism reveals therapeutic targets treating both acute lung injury and high dose IL-2 toxicity in cancer.
Cellular stress mediated by the local microenvironment regulates the homeostatic loss of tissue-resident memory CD8 T cells in different compartments of the lung
Authors: Hayward Sarah L. , Scharer Christopher D. , Takamura Shiki , Kumar Gaurav , Boss Jeremy M. , Kohlmeier Jacob E. 
Published: January 2023

Abstract
Lung-resident memory T cells (lung TRM) are important for protective heterosubtypic immunity against influenza viruses; however, the efficacy of this cellular immunity wanes over time. To better understand the mechanisms leading to the decline of protective cellular immunity, we investigated the long-term maintenance of airway and parenchyma lung TRM. In contrast to circulating flu-specific memory T cells and TRM in other tissues, lung TRM numbers in the airways and parenchyma steadily decline for several months following influenza infection. All flu-specific lung TRM decline at similar rates regardless of expression of the tissue residency markers CD69 and CD103. Parabiosis experiments demonstrate that established lung TRM in the airways or parenchyma do not freely interchange with circulating memory T cells. Thus the gradual loss of these cells is likely due to increased cell death in the tissue. In support of this, airway and parenchyma lung TRM show increased apoptosis compared to systemic flu-specific memory T cells during homeostasis. To identify epigenetic and transcriptional programs specific to lung TRM, we performed parallel RNA-seq and ATAC-seq on flu-specific memory CD8 T cells from the airways, lung parenchyma, and spleen. We found that lung TRM populations have increased expression of genes associated with cellular stress, and airway TRM in particular have a signature associated with amino acid starvation and ER stress. Finally, intratracheal transfer experiments determined that the airway environment was sufficient to program the lung TRM transcriptional signature. Together, these data show that the decline of lung TRM is a direct result of cellular stresses driven by the lung microenvironment.
The natural suppression of respiratory CD8 T cell memory by monocyte-derived dendritic cells (moDC)
Authors: Reagin Katie L. , Slade Chris D. , Klonowski Kimberly D. 
Published: January 2023

Abstract
Heterosubtypic CD8 T cell immunity can protect mice and humans against reinfection with highly mutagenic viruses like influenza. Studies in mice attribute this protection to CD8+ tissue resident memory cells (TRM) residing in the lung and airways. However, both live attenuated influenza vaccines and natural influenza infection fail to protect humans from seasonal influenza. Interestingly, unlike TRM derived from other mucosal infections, the majority of TRM in the lung parenchyma are devoid of CD103 and CD69 expression, suggesting bona-fide TRM are either not generated or stably maintained. Using a mouse model, we show respiratory infection evokes the selective migration of CD11c+CD11b+CD103+CCR2+PD-L1hi moDCs to the lung-draining lymph nodes in response to a variety of pathogens (influenza, RSV, VSV) 72–96 hours post infection, coordinate with T cell priming. These moDC develop in the context of systemic and intranasally VSV challenged animals and influenza infected mice, but only emigrate from the bone marrow after respiratory infection. Lymph node immigration is coordinate with regional CCL2 expression. Importantly, the selective depletion of moDCs using CCR2-DTR bone marrow chimeras numerically enhances CD103+CD69+ TRM formation and increases local IFN-ɣ and TNF-α after heterologous virus challenge. In summary, our studies show that moDCs selectively suppress TRM formation after respiratory infection. Ongoing studies seek to understand the molecular mechanism(s) used by moDCs to alter CD8 T cell programming and TRM development.
Comparison of two platforms for the measurement of fg/mL concentrations of protein biomarkers using single molecule arrays and digital ELISA: the benchtop reader Quanterix SR-X™, and the fully automated analyzer HD-1™
Authors: Lambert Jeremy , Vonk Ariel , Mendes Muriel , Song Linan , Svancara Danielle , Baig Shazia , Rissin David , Nie Shuai , Hanlon David , Patel Purvish , Duffy David , Shan Dandan , Chang Lei , Johnson Joe 
Published: January 2023

Abstract
Digital ELISA (Enzyme Linked Immunosorbent Assay) based on single molecule arrays (Simoa) has improved sensitivity of traditional ELISA from picomolar (10−12M) to femtomolar (10−15M), increasing the quality and quantity of biomarkers that can be measured for health and disease. Digital ELISA counts signal generated from single immunocomplexes formed on superparamagnetic beads confined in arrays of femtoliter-sized wells in which fluorescent molecules are highly concentrated.
Quanterix has developed digital ELISA assays in a fully-automated instrument (Simoa HD-1 Analyzer), ideal for use in pharmaceutical companies, drug discovery, clinical research and other areas necessitating full automation and high throughput. Recent advancements in the Simoa technology and workflow have been integrated into the new SR-X benchtop reader, with a smaller footprint and more flexible workflow. Operators prepare assays in microtiter plates at the bench in a semi-automated format similar to traditional ELISA, with the notable exception that plates are preserved by drying after assay completion, and can be read immediately or the next day.
We present results comparing performance of the following Simoa assays on SR-X to HD-1: PSA; HIV p24; Tau; Neurofilament-light; PD-L1; TNFa; IL-10; IL-17A; IL-6 and a neurology multiplex panel consisting of NF-L, tau, GFAP and UCH-L1. Measured sample levels correlated with R2 values from 0.96 to 1.00, with average LOD and LLOQ within 1.4 and 1.5 fold of HD-1, respectively. Inter-assay precision ranged from 4.0 to 11.2% CV across assays. Operators tested full-plates from start to finish within 1 – 2 hours (1/2 hour hands on time) and a read time of 2 hours (5 minutes hands on time).