bluuebunny/crossref_metadata_2025_split huggingface dataset:
UDP-glucose Regulates Dendritic Cell Mitochondrial Respiration via a Nitric Oxide-dependent Mechanism
Authors: Vagher Bay L , Amiel Eyal 
Published: August 2023

Abstract
Dendritic cells (DCs) are key innate immune cells specialized in fine tuning an immune response. Activation of DCs through Toll-Like Receptors (TLRs) is accompanied by a metabolic switch to increased use of glycolytic metabolism fueled by carbohydrates glucose and glycogen. Our lab has shown that DCs require glucose and glycogen to perform their immune effector functions including: production of cytokines and inflammatory mediators, antigen presentation to T cells, and initiation of the T cell-mediated immune response. Recent studies in inflammatory macrophages showed a secondary metabolite in the glycogen synthetic pathway, uridine diphosphate glucose (UDP-glucose), is upregulated through TLR4 stimulation. UDP-glucose exits the cell and signals in an autocrine manner through its specific purinergic receptor P2Y14. Additionally, P2Y14R signaling modulates the expression of proinflammatory pathways including upregulation of inducible nitric oxide synthase (iNOS). Activated bone marrow-derived DCs (BMDCs) upregulate iNOS and produce nitric oxide (NO), which has pleiotropic effects, including inhibiting mitochondrial respiration. Therefore, we hypothesized that the UDP-glucose/P2Y14R/iNOS axis regulates BMDC metabolism, which has further ramifications on DC cell survival and proinflammatory phenotypes. To investigate the impact of UDP-glucose as an autocrine signaling molecule, we characterized proinflammatory gene transcript and protein levels, as well as the functional outcomes of increased iNOS protein in the cell. These studies showed that DCs use the UDP-glucose/P2Y14R/iNOS axis to bolster inflammatory phenotypes including inhibition of mitochondrial respiration leading to exacerbated cell death.
Supported by grants from NIH (R01 AI158710-01, 1R21AI135385-01A1) and UVM (COBRE VCIID Funding Support and Pilot Project)
Gene-dose-dependent roles of DEFA1A3 in the neutralization of uropathogenic Escherichia coli
Authors: Canas Jorge J , Schwaderer Andrew L , Hains David S 
Published: August 2023

Abstract
Genetic polymorphisms of the human genome provide information about disease susceptibility and autoimmunity. Genetic variations involving the innate immune system have the potential to improve the effectiveness of diagnosis and treatment of urinary tract infections in children and adults. DNA copy number variations (CNVs) of DEFA1A3, which encodes for antimicrobial peptides α-defensin 1–3, have been associated with recurrent UTI risk in children with vesicoureteral reflux (VUR). α-Defensins 1–3 are pleiotropic antimicrobial peptides with bactericidal and immunomodulatory properties. Due to the absence of a gene ortholog in mice, traditional murine models to study the effect of DEFA1A3 CNVs are lacking. Our objective is to explore the gene-dosage pleiotropic effects of the DEFA1A3 locus by utilizing a UTI-challenged transgenic mouse model expressing variable copies of the human gene. To model murine UTIs, uropathogenic E. coli (UPEC) pyelonephritis strain (CFT073) or vehicle volume was injected transurethrally into DEFA4/4, DEFA4/0, and DEFA0/0 mice. Urinary tract organs were analyzed for quantifiable bacterial growth, immune cell frequencies, and antimicrobial response gene expression array. Kidney bacterial clearance effects are proportional to gene-dosage; gradually decreasing colonization in mice between DEFA4/4 (71%), DEFA4/0 (57%), and DEFA0/0 (43%) genotypes. The kidney immune cells displayed differential mRNA expression of pro-inflammatory genes Il1β and Il6 genes as well as phagocytic genes Rac1 and Lyz2 between the infected mice genotypes. Our findings support pleiotropic gene-dosage protective roles of the human DEFA1A3 gene in a murine model of UTI-induced pyelonephritis.
Supported by the grants from NIH (R01 DK117934 & R01 DK106286)
IL-33 affects the IL-4-induced development of group 2 innate lymphoid cells in neonatal lungs
Authors: Huang Hui-Yu , Shieh Chi-Chang 
Published: August 2023

Abstract
Abundant IL-33 is produced by alveolar type 2 cells during lung alveolarization phase in the neonatal stage. It promotes Th2 response and may facilitate ILC2 development with IL-4 stimulation. IL-4 has been found to induce IL-10+ ILC2s in the lung environment, however, it is still unknown whether ILC1/2/3 transdifferentiation and IL-10+ ILC2 functions in neonatal lungs are regulated by IL-33. Therefore, we aimed to investigate the regulation role of IL-33 in lung ILC development during neonatal period. To identify the key cytokines under IL-33 regulation in neonatal lungs, we used antibody staining to characterize ILC1/2/3 and measured the cytokines relevant to ILC2 functions in WT and IL-33−/− mice. We found that while lung ILC2s increased from postnatal day (PND) 14 to 28 in WT mice, ILC1s and ILC3s decreased in the same period. However, we found that there were significantly decreased ILC2s on PND 21 in the lungs of IL-33−/− mice when compared with WT mice. We also found that increase in IL-33, IL-4, IL-10 on PND 28 in the lungs of WT mice, which was not apparent in IL-33−/− mice. The sorted neonatal lung ILC2s may secrete more IL-10 after stimulated with IL-4 and IL-33 when compared with adult lung ILC2s from either WT or IL-33−/− mice. Moreover, in vitro cultured ILC populations from IL-33−/− mice after stimulated with IL-4 and IL-33 may enhance ILC1/3 transdifferentiate into IL-10+ ILC2s. Our results showed that IL-33 contributes to ILCs differentiate towards IL-10+ ILC2 through IL-4 and IL-33 stimulation during lung alveolarization phase. As IL-10+ ILC2s have been found to play a regulatory role in allergy, the development of IL-10+ ILC2s during neonatal period may be critical for asthma prevention.
Dysregulation of fetal-derived cochlear resident tissue macrophages mediates CMV-associated sensorineural hearing loss
Authors: Otsuka Kelly , Scott Boston , Park Albert , Beaudin Anna E 
Published: August 2023

Abstract
Cytomegalovirus (CMV) is the most common congenital viral infection in the developed world and the leading cause of non-genetic sensorineural hearing loss (SNHL) in children. Congenital CMV causes progressive SNHL long after acute infection has resolved, suggesting a lasting effect on the developing host immune response in the cochlea. Using fate-mapping models, we investigated the contribution of fetal-derived resident tissue macrophages (RTMs) to normal cochlear development and function, and their role in mediating CMV-associated SNHL. We uncovered dual contribution of fetal-derived cochlear RTMs (F4/80hiCD11blo) from yolk sac (YS) and fetal liver (FL) hematopoiesis at birth. YS-derived RTMs naturally recede postnatally after cochlea development, but remain in key tissues, such as the spiral ligament and stria vascularis, through adulthood. Depletion of fetal-derived RTMs by anti-CSF1R blocking Ab led to SNHL and impairments to cochlear architecture. Similarly, embryonic deletion of YS-derived RTMs by induced deletion of CSF1R leads to SNHL at high frequencies. During acute CMV infection, RTMs expand and become transcriptionally dysregulated in developmental programs, including tissue remodeling, chemorepulsion, otic vesicle morphogenesis, and FGF signaling. Importantly, analysis of CSF1R-Cre labeling suggests YS-derived RTMs retract in response to CMV infection, suggesting RTM expansion is caused by infiltration of BM-derived macrophages. Ongoing work examines spatial distribution of fetal-derived cochlear RTMs, transcriptional differences between YS, FL, and BM-derived RTMs in response to CMV, and how replacement of fetal-derived RTMs with BM-derived RTMs affects tissue function and response to CMV.
Supported by grants from NIAID (T32 5T32AI138945) and University of Utah molecular medicine and immunology, inflammation and infectious disease (3i) initiative
Pseudomonas aeruginosa pulmonary infection results in S100A8/9-dependent cardiac dysfunction
Authors: Rajaram Murugesan , Kumar Naresh , Pestrak Matthew , Wu Qian , Ahumada Omar , Saljoughian Noushin , Shukla Rajnikant , Mitchem Cortney , Nagareddy Prabhakara , William Lafuse , Ganesan Latha , Wozniak Daniel J 
Published: August 2023

Abstract
Pneumonia in ICU patients carries a bleak prognosis with high risk for cardiac dysfunction, largely due to cardiac infection and inflammation. Pseudomonas aeruginosa (P.a.) infection accounts nearly 20% of all cases of hospital acquired pneumonia with mortality rates &gt;30%. P.a. infection induces a robust inflammatory response, which ideally enhances bacterial clearance. Unfortunately, excessive inflammation can also have negative effects, and often leads to cardiac dysfunction with associated morbidity and mortality. However, it remains unclear how P.a. lung infection causes cardiac dysfunction. Using a murine pneumonia model, we found that of P.a. infection of lungs led to severe cardiac left ventricular dysfunction and electrical abnormalities. More specifically, we found that neutrophil recruitment and release of S100A8/A9 in the lungs activates the TLR4/RAGE signaling pathways, which in turn enhance systemic inflammation and subsequent cardiac dysfunction. Paradoxically, global deletion of S100A8/A9 did not improve but aggravated cardiac dysfunction and immortality likely due to uncontrolled bacterial burden in the lungs and heart. Our results indicate that P.a. infection induced release of S100A8/9 is double-edged, providing increased risk for cardiac dysfunction yet limiting P.a. growth.
Funding: NIH grants R01-AI 146252 and R21-AG 127783
Funding: NIH grants R01-AI 146252 and R21-AG 127783
Disease associated monocytes contribute to the inflammatory milieu in hidradenitis suppurativa and have dysregulated expression of transposable elements
Authors: Dimitrion Peter Michael , Hamzavi Iltefat , Yin Congcong , Loveless Ian , Toor Jugmohit , Sulbedi Kalpana , Khalasawi Namir , Miller Angela , Huggins Richard , Adrianto Indra , Veenstra Jesse , Vellaichamy Gautham , Hans Aakash , Daveluy Steven , Athar Mohammad , Liao Wilson , Lim Henry , Ozog David , Zhou Li , Mi Qing-Sheng 
Published: August 2023

Abstract
Hidradenitis suppurativa (HS), an inflammatory skin-disease, manifests as painful lesions in intertriginous skin. Despite their central role in innate immunity the role of monocytes in HS is largely unknown. Here we utilized cytometry by time-of-flight (CyTOF), imaging CyTOF (IMC), cellular indexing of transcriptomes and epitopes by sequencing (CITEseq), spatial transcriptomics (ST), and our recently developed database (HSOmicsDB) to characterize the systemic and cutaneous immune response in patients with HS. CITEseq found differential genes in circulating immune cells from HS patients enriched for cellular activation and extravasation. CyTOF immune profiling of fresh whole-blood samples found decreased frequency of classical Mos (C.Mos), which had increased expression of skin homing chemokine receptors. IMC found C.Mos infiltrated lesional dermis and tunnel regions, but not perilesional (PL) skin. Patients with HS showed increased proportions of activated C.Mo subclusters and a population of SOD2+ C.Mos expressing genes downstream of viral sensing pathways. SOD2+ C.Mos expressed high levels of neutrophil attracting chemokines. ST found SOD2+ C.Mos localized to regions of active inflammation and dermal tunnels. HSOmicsDB showed Mos and Macs expressed SOD2+ C.Mo genes, which were enriched in lesions, but not PL skin. Finally, we show that SOD2+ C.mos in HS have dysregulated expression of transposable elements (TEs), which may be activating viral sensing mechanisms in patients with HS. Together our data shows monocytes in HS function to recruit neutrophils to lesional HS skin, and provides the first evidence implicating TEs as disease promoting factors in HS by leading to C.Mo dysregulation.
Supported By NIH/NIAMS, R01AR078688-01A1 and R21AR079089-01A1
Neonatal bacilllus Calmette-Guerin (BCG) administration did not limit influenza A virus replication or clinical disease in pigs
Authors: Loving Crystal L , Hill Dana , Byrne Kristen 
Published: August 2023

Abstract
A potential immunomodulation mechanism for improving disease resilience is innate training, which relies on epigenetic modifications of immune cells for a heightened (ie, trained) response upon repeated microbial stimulation. Prior experiences indicate bacillus Calmette-Guerin (BCG) exposure increases cytokine production upon secondary exposure to microbe-associated molecular patterns, and epidemiological data suggest improved disease resistance in humans administered neonatal BCG. Neonatal piglets received BCG or mock inoculum via intravenous (IV) route and were subsequently exposed to influenza A virus (IAV) inoculated piglets. Body temperature was recorded and nasal swabs collected through the acute phase of viral infection. At necropsy, gross lung pathology was assessed. In addition, prior to IAV challenge, peripheral blood mononuclear cells (PBMC) and monocytes were isolated from mock and BCG inoculated animals and stimulated in vitro with LPS to evaluate in vivo induction of a trained phenotype via cytokine production and gene expression. PBMC were stimulated with PPD antigen to assess antigen-specific T cell responses to BCG inoculation. PPD-specific IFNγ production by PBMCs was minimal; yet upon in vitro LPS exposure, monocytes isolated from IV BCG treated piglets produced more IL-1β and TNF cytokine than monocytes from mock pigs (ie, trained phenotype). However, no significant differences in clinical presentation or nasal and lung IAV titers were detected. BCG administration induced a trained phenotype in porcine peripheral monocytes, it did not translate into a significant alteration in respiratory viral infection.
USDA-ARS-NIFA 2019-07511
Sex differences in ILC2 function in influenza result from imprinted donor sex intrinsic factors independent of host sex in competitive female-male bone marrow chimeras
Authors: Williams Abigael Parrish , Turner Sean , Chlebicz Magdalena , Miller Reegan , Roe Mandi , Kadel Sapana , Kovats Susan 
Published: August 2023

Abstract
Historical influenza pandemics and the SARS-CoV-2 pandemic revealed gender differences in morbidity of respiratory virus infections. In human or murine influenza A virus (IAV) infection, females of reproductive age generate stronger innate and adaptive antiviral responses leading to immunopathology, while elderly males often experience more severe disease. Sex hormone levels play a role in this disparity, yet little is known about cell intrinsic regulation of immune cells by estrogen or androgen receptors (AR). Lung resident type 2 innate lymphocytes (ILC2s) express high levels of AR, a nuclear hormone receptor that regulates chromatin via epigenetic modifications. While female lungs contain higher ILC2 numbers in homeostasis, we showed that ILC2s in females were preferentially functionally suppressed during IAV infection, whereas ILC2s in males retained robust type 2 function, including IL-5 production. Here, we determined if sex differences in pulmonary ILC2s result from extrinsic factors present in host environments or from imprinted donor-intrinsic factors. We set up competitive bone marrow chimeras by transferring a mix of female and male lineage-depleted bone marrow cells into either male or female hosts. Female hosts harbored more pulmonary ILC2s than male hosts, suggesting the extrinsic sex hormone environment regulates ILC2 numbers. Upon IAV infection, an increased fraction of ILC2s from male donors retained high IL33R expression and produced IL-5, regardless of host sex, suggesting donor sex-intrinsic factors regulate the canonical ILC2 functional response. We are now testing the hypothesis that androgens/AR activity epigenetically imprint genes regulating type 2 function in male ILC2s upon sexual maturity.
Single-cell cloning of NK cells from rhesus macaques: a new methodology for in-depth analysis of MHC-E-dependent adaptive functions
Authors: Rascle Philippe M , Jones Rhianna , Lee Esther , Terry Karen , Jost Stephanie , Reeves R Keith 
Published: August 2023

Abstract
Natural killer (NK) cells provide rapid and robust responses against a wide range of viral infections. More recently, multiple studies have shown that subsets of NK cells are also capable of antigen-specific recall to eliminate infected cells. In humans and nonhuman primates (NHP), antigen-specific memory NK cell responses can at least be partially delineated by expression of NKG2C, a ligand for MHC-E. Due to MHC-E limited polymorphism, MHC-E-presented viral antigens may induce universally antigen-specific memory NK cells.
To study antigen-specific NK cells in NHP, we developed a new in vitro method to clonally expand single rhesus macaque NK cells (rhNKCL) from blood and spleen. To define the effector functions of rhNKCL, we used a calcein acetoxymethyl ester (CAM) cytotoxicity assay and intracellular staining assay in combination with K562 cell lines or autologous BLCL as targets. We also identified, by in silico methodology, peptides from influenza, SIV, and SARS-CoV-2 viruses predicted to display strong binding to MHC-E.
Our procedure showed for the first-time clonal expansion of single rhesus macaque NK cells. We observed a conserved NHP NK cell phenotype of CD3− NKG2A/C+ CD16+ CD56+ following expansion with expression of additional NK cell surface receptors CD8, NKp80, and NKG2D. Upon stimulation with K562 cells, rhNKCL effector functions (cytotoxicity, CD107a, IFNγ, TNFα) were like primary NK cells.
Altogether, this new method of single-cell cloning in rhesus macaques could provide key information about antigen-specific NK cells, including epigenetic, metabolic, transcriptional, and phenotypic profiles, which could be exploited to further enhance antiviral NK cell activity in future vaccine strategies or cell therapies.
Supported by grants from NIH (R01 AI158516, R01 DK130472, R01 AI161010, NIH-funded BEAT-HIV Martin Delaney Collaboratory to cure HIV-1 infection by Combination Immunotherapy UM1 AI164570).
The Immunoregulatory Role of CMKLR1 within the CNS During WNV Encephalitis
Authors: Kwok Keven , Durrant Douglas M 
Published: August 2023

Abstract
West Nile Virus (WNV)-induced neuroinvasive disease (WNND) has been a public health concern in North America since its introduction in 1999. In recent years, severe neurological illness has been reported much more frequently, together with neuromuscular manifestations. Previous studies have shown that dendritic cells (DCs) within the central nervous system (CNS) are required to restrict WNV replication and immunopathology by regulating anti-viral T-cell responses. In this study, we examined the role of CMKLR1 within the CNS during WNV encephalitis. CMKLR1-expressing microglial cells and CNS-infiltrating myeloid DCs have been shown to have an inflammatory role during autoimmune neurological disorders, yet their contribution to WNND remains unknown. In addition, chemerin, the ligand for CMKLR1, was upregulated in the CNS of WNV-infected mice. To evaluate the role of CMKLR1, we infected CMKLR1-deficient (CMKLR1−/−) and wild-type (WT) mice with WNV. WNV-infected CMKLR1−/− mice exhibited increased susceptibility to WNV, loss of virologic control, and more severe clinical signs of disease. Proinflammatory chemokine expression, including CCL5, CXCL9, and CXCL10, was significantly decreased in the absence of CMKLR1 suggesting reduced leukocyte trafficking into the CNS. For future studies, we will determine the numbers and type of infiltrating leukocytes and their role in virologic control within the WNV-infected CNS. Taken together, these data will help clarify the immunoregulatory role of CMKLR1 during WNV encephalitis.
NIH-SC3GM130472
The entry receptor LDLRAD3 is critical for Venezuelan equine encephalitis virus pathogenesis and neuroinvasion in mice
Authors: Kafai Natasha M , Janova Hana , Alippe Yael , Muraro Stefanie , Diamond Michael S 
Published: August 2023

Abstract
Alphaviruses are enveloped positive-stranded RNA viruses of the Togaviridae family. Venezuelan equine encephalitis virus (VEEV) is an encephalitic alphavirus responsible for epidemics in equine and human populations across the Americas. Its potential for use as an aerosolized bioterrorism weapon and the demonstrated risk it poses in research settings highlight the need for countermeasures. VEEV pathogenesis is characterized by a systemic inflammatory response and central nervous system (CNS) infection, which can result in severe neurological complications and sequelae. Our laboratory identified a novel entry receptor for VEEV, low-density lipoprotein receptor class A domain containing 3 (LDLRAD3). Ldlrad3−/− mice demonstrate reduced, but not abrogated, VEEV infection in target tissues and are resistant to disease and lethality. Here we define a critical role for LDLRAD3 in controlling VEEV tissue tropism and pathogenesis by studying the kinetics of infection in wild-type and Ldlrad3−/− mice. Using detailed analyses of viral RNA levels, immunohistochemistry, flow cytometry, and primary neuronal cultures, we observed that VEEV infects LDLRAD3-expressing cell types as early as 6 h after inoculation and once in the CNS, preferentially targets neurons, and not glial cells, in a LDLRAD3-dependent manner. Overall, our studies define the role of LDLRAD3 in VEEV tropism and how LDLRAD3-dependent infection of LDLRAD3 contributes to the host inflammatory responses against VEEV. Finally, our studies demonstrate LDLRAD3-independent infection in Ldlrad3−/− mice, which suggests the existence of alternate receptors for VEEV. These findings may be relevant for the generation of novel receptor-targeted antiviral therapies.
NIAID F30AI164842
The mutation PTPN6Ala455Thr causing pulmonary emphysema affects B cell populations and causes spontaneous pulmonary tertiary lymphoid tissue formation
Authors: Pineault Marie , Bouffard Gabrielle , Milad Nadia , Tremblay Félix , Lechasseur Ariane , Routhier Joanie , Aubin Sophie , Beaulieu Marie-Josée , Bossé Yohan , Maltais François , Morissette Mathieu C 
Published: August 2023

Abstract

Rationale
A new mutation causing severe pulmonary emphysema was recently discovered in a French-Canadian family. The mutation is located in the PTPN6 gene (PTPN6Ala455Thr) leading to a reduction in the activity of SHP-1, a phosphatase that regulates pathways associated to B cells. We aimed to characterize B cell abnormalities associated with aging in both humans and mice carrying this mutation.

Methods
Circulating B cell populations in humans carrying the PTPN6Ala455Thr mutation were analyzed by flow cytometry and immunoglobulin levels were measured in serum. In mice carrying the same mutation (Ptpn6Ala457Thr) aged up to 12 months, B cell populations were analyzed by flow cytometry and lung tissue histology performed. Mice were immunized with the T cell-independent antigen and NP-specific antibodies were measured in plasma.

Results
Human carriers of the PTPN6Ala455Thr mutation had reduced IgG1 and IgG4, and increased IgG3 levels as well as a lower ratio of naive B cells/ isotype switched memory B cells compared to controls. Aging Ptpn6Ala457Thr mice developed spontaneous pulmonary tertiary lymphoid tissues and exhibited higher proportion of B-1 cells, age-associated B cells and germinal center B cells along with lower B-2 cells in the lung and spleen. Ptpn6Ala457Thr mice had lower levels of NP-specific IgG1 antibodies compared to the wild-type group.

Conclusion
SHP-1 appears to affect aging of B cells as the PTPN6Ala455Thr mutation leads to changes in B cell populations associated with age, tertiary lymphoid tissue formation and an alteration of immunoglobulin levels. The link between B cell abnormalities, aging and emphysema requires further investigation.
Funding:CIHR, FRQS

CIHR, FRQS