bluuebunny/crossref_metadata_2025_split huggingface dataset:
The lung hematopoietic niche supports bone marrow-independent hematopoiesis and contributes to local eosinophil expansion during allergic inflammation.
Authors: Jeong Brian M. , Coden Mackenzie , Doan Ton C. , Chiarella Sergio , Rodriguez Raul , Berdnikovs Sergejs 
Published: January 2023

Abstract
Bone marrow is classically viewed as the major source of differentiated leukocytes infiltrating tissues from circulation. Recent evidence shows that the lung can potentially function as a hematopoietic organ on its own. This is a novel concept and tissue factors driving in situ leukocyte production are not yet well understood. In naïve murine lungs, we detected a small population of Lin−CD34+c-Kit+ common myeloid progenitors (CMPs) by flow cytometry and single cell RNA-seq analysis. Males had 0.087±0.06% CMPs, while females had a larger population of 0.18±0.08%, out of CD45+ cells. In a 3 challenge in vivo ovalbumin model of allergic inflammation, we observed a six-fold expansion of CMPs in the lung tissue and the appearance of Siglec-F+CD34+IL-5Rα+ eosinophil progenitors (EoPs) (1.505±0.6% of CD45+). To illustrate the hematopoietic potential of the lung independent of bone marrow recruitment, we cultured naïve lung single cell suspensions ex vivo in presence of SCF, FLT3 and IL-5. This was sufficient to yield up to 20% mature eosinophils, which is comparable to tissue eosinophil levels in models of allergic inflammation. We further tested whether Tenascin-C (TNC), a known component of bone marrow hematopoiesis, may equally support the hematopoietic niche in the lung. TNC is a provisional matrix glycoprotein that strongly associates with epithelial remodeling and eosinophil accumulation in asthma. TNC−/− mice had a significant defect in CMP and EoP expansion in a murine model of asthma and ex vivo IL-5 lung cultures. Administration of recombinant TNC rescued normal lung eosinophilopoiesis. These results suggest the existence and significant potential of the lung hematopoietic niche to supply effector cells during inflammation.
Resistin-like molecule-β negatively regulates chronic features of fungal asthma
Authors: Samarasinghe Amali , LeMessurier Kim , Palipane Maneesha , Tiwary Meenakshi , Gavin Brian 
Published: January 2023

Abstract
Severe asthma with fungal sensitization (SAFS) is a subtype of asthma with high incidence in which patients are often therapy-resistant and have chronic disease features such as airway wall remodeling. Shared embryonic, immunologic, and microbiomic lineages of the gut-lung axis should be considered in disease pathogenesis investigations. A resistin family hormone, RELM-β, has been implicated in regulation of mucus production, immune responses, and bacterial colonization in the gut. RELM-β has been previously shown to increase macrophagic inflammation, goblet cell (GC) metaplasia, and fibrosis in mouse models. Since fungal asthma has unique early- and late-phase responses, we investigated the functions of RELM-β in our well-established model of SAFS utilizing Retnlb-null (KO) mice. Characteristics of SAFS developed in KO mice with some differences from wild-type (WT) controls. Kinetics of airway inflammation were delayed in KO although tissue inflammation was similar to WT. GCs interspersing the bronchial epithelia were similar, however, hyperexpression of Muc5ac and Muc5b in KO mice suggests of an altered mucin composition. Subepithelial fibrosis and airway resistance were markedly increased at late timepoints in KO mice compared to WT, indicating that RELM-β inhibits lung fibrosis. RELM-β may function as a homeostatic regulator of B cells since naïve KO mice had elevated systemic IgA. Related molecule, RELM-α, did not increase in KO mice although antimicrobial peptide Camp, Il13, and Tlr4 were hyperexpressed in lungs of these mice. Cumulatively, these data suggest that RELM-β is a regulator of allergic responses and that these effects may occur through direct/indirect mechanisms.
Therapeutic blockade of CD3-mediated signaling ameliorates memory Th2-driven allergic asthma
Authors: Gracias Donald Tom , Mehta Amit K. , Croft Michael 
Published: January 2023

Abstract
Allergic asthma is an inflammatory disease of the lungs characterized by Type 2 helper T cell (Th2)-driven immune responses to inhaled allergens. Memory Th2 cells are a major driving force behind asthma, secreting cytokines like IL-4, IL-5, and IL-13 as well as persisting in the host long after allergen encounter. The selective removal of pre-existing memory Th2 cells could be a key process resulting in enhanced tolerance. One such method could involve targeting CD3 through the use of monoclonal anti-CD3 antibodies, which have been shown to successfully induce tolerance in animal models of autoimmune diseases and have also shown promise in clinical trials. We tested blocking CD3 complex signaling in a HDM model of allergic inflammation, to determine if we could induce airway tolerance. Importantly, pre-sensitized C57BL/6J mice displayed strongly ablated lung inflammation and Th2 responses when treated at the time of memory T cell reactivation with F(ab′)2 anti-CD3 antibody. This treatment was found to specifically inhibit HDM-specific CD4+ T cell proliferation and Th2 cytokine production. Interestingly, mice treated with anti-CD3 also showed an increased frequency of Treg cells, improving the Treg:Th2 cell ratio in favor of airway tolerance. Additionally, a significant reduction in airway inflammation and Th2 responses was maintained with therapeutic anti-CD3 antibody even when given several months later during a further memory T cell recall response to allergen. These data suggest that blocking signaling mediated by the CD3 complex could be employed to inhibit memory recall responses to complex allergens thus promoting airway tolerance.
Response Gene to Complement-32 Expression Is Upregulated in Lupus T cells and Promotes IL-17A Expression
Authors: Rus Violeta , Tatomir Alexandru , Nguyen Vinh , Rus Horea G. 
Published: January 2023

Abstract
RGC (Response Gene to Complement)-32 is a cell cycle regulator expressed in normal tissues, tumors and a variety of cell lines. RGC-32 is preferentially upregulated in murine Th17 cells and promotes their differentiation. Patients with Systemic Lupus Erythematosus (SLE) display increased serum levels, expanded frequency of IL-17 producing cells. Whether RGC-32 plays a role in human Th17 differentiation pathway and in Th17 abnormalities in lupus patients has not yet been investigated. RGC-32 mRNA expression was determined in CD4+ T cells stimulated under Th0, Th1, Th2, Th17 and Treg conditions and the effect of RGC-32 overexpression and silencing under Th17 conditions was determined by nucleofection. RGC-32 expression was evaluated by RT-PCR, flow cytometry and cDNA Array in PBMC from lupus patients vs controls. RGC-32 mRNA expression was upregulated by TCR stimulation and TGFβ, and was more robust under Th17 (3.2 ± fold) and Treg (2.6 ± 0.8 fold) vs. Th1 (1.3 ± 0.4 fold) and Th2 (1.8 ± 0.1 fold) conditions. Overexpression or silencing of RGC-32 in CD4+ T cells upregulated, respectively downregulated IL-17A transcript levels and protein secretion. RGC-32 mRNA and protein level was significantly increased in CD19+ B cells and CD3+ T from lupus patients compared to controls. These results suggest that RGC-32 promotes the differentiation of human Th17 cells. Furthermore, T cells from patients with SLE exhibit increased expression of RGC-32 compared to controls. These data support the idea that RGC-32 signaling may enhance disease expression in SLE by promoting abnormalities in the Th17 pathway and provide a compelling rationale for further investigating the therapeutic potential of blocking RGC-32 in SLE.
Enhanced interferon gamma response contributes to disease remission in Rheumatoid Arthritis.
Authors: Sharma Vishal , Pope Brandon J. , Bolland Molly , Reynolds Richard , Sun Dongmei , Bridges S. Louis , Raman Chander 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is an autoimmune disease with debilitating bilateral inflammation in multiple joints. Although newer biologics are very effective in managing RA, critical gaps in immunopathogenic mechanism remain. We recently showed that increased expression of the interferon gamma (IFN-γ) receptor in PBMC associated with RA and increased radiographic severity. We hypothesized that enhanced IFN-γ signaling in specific subpopulations of mononuclear cells from patients with RA contributes to disease severity. To test this, we collected PBMC from healthy controls (HC) and RA patients in remission, low disease and moderate disease severity as per American College Criteria. As an autoimmune disease control, we compared PBMC from treatment naïve multiple sclerosis (MS) patients. We quantitated classical (JAK/STAT) and non-classical (ERK/MTOR) IFN-γ induced signaling in subpopulations of T and B cells within PBMC using phospho-flow cytometry. For analysis we utilized t-SNE and SPADE data visualization and clustering tools to interrogate changes in activation and marker expression within and between T and B cell populations. We observed IFN-γ-induced activation of STAT1 in CD4+ naïve and central memory T-cells from was greatest in RA patients in remission and lowest in individuals with moderate disease. IFN-γ induced activation of STAT1 in all CD8+ T-cell sub-populations of RA were similar to that from HC. Furthermore, the activation of STAT1 in all CD4+ T-cell sub-populations in PBMC from MS patients very generally greater than from RA patients and HC. From our results we infer that that while expression of IFNGR correlates with disease in RA, gain in response to IFN-γ is indicative of treatment response and RA remission.
Natural killer cell dysregulation underlies atopic dermatitis
Authors: Mack Madison R. , Brestoff Jonathan R. , Niu Hiaxia , Whelan Timothy M. , Oetjen Landon K. , Bodet Nancy D. , Wang Fang , Xu Amy Z. , Park Eugene , Margolis David J. , Yokoyama Wayne M. , Kim Brian S. 
Published: January 2023

Abstract
Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disease characterized by dry, itchy rashes that affects 5–10% of the US adults. AD commonly presents in childhood and is known to be complicated by reduced antiviral skin immunity. Although classically thought to be driven by adaptive TH2 cells, recent work has demonstrated that dysregulation of innate immune cells such as group 2 innate lymphoid cells (ILC2s) and basophils can promote its pathogenesis. However, how these innate immune cells become dysregulated remains unclear. Using paired RNA-Seq of lesional and non-lesional skin in patients, we identified an enrichment for natural killer (NK) cell-associated gene sets in AD skin lesions. NK cells are well-recognized innate effector cells for antiviral and anticancer immunity. However, their role in AD has not been established. We utilized unbiased multidimensional analysis of gene expression and flow cytometry data to identify underlying NK cell dysregulation in AD patients. Further, we demonstrate that NK cells negatively regulate type 2 inflammation in a mouse model of AD. Collectively, these studies reveal previously unrecognized functions of NK cells in cutaneous immune homeostasis.
The expression of CD22 and CD72 on B cells negatively correlate with IgE level in eosinophilic chronic rhinosinusitis with nasal polyps
Authors: Wang Hai , Song Jia , Wang Zhe-zheng , Ruan Jian-wen , Liu Zheng 
Published: January 2023

Abstract

Background
CD22 and CD72 have been demonstrated to act as B cell inhibitory receptors, which suppress B cells activation. However, the roles of CD22 and CD72 on B cells in nasal polyps (NPs) are largely unknown.

Objective
To explore the expression of CD22 and CD72 on B cells and the association with immunoglobulins (Igs) in NPs.

Methods
The expression levels of CD22 and CD72 was measured by RT-PCR, immunohistochemistry, immunofluorescence and flow cytometry. Igs levels in NPs were quantified by Bio-Plex assay or the UniCAP system. Computed tomography scan and symptom severity were graded.

Results
The mRNA expression of CD22 and CD72 were elevated in eosinophilic chronic rhinosinusitis with nasal polyps (Eos CRSwNP) patients compared with non-eosinophilic chronic rhinosinusitis with nasal polyps (non-Eos CRSwNP) and controls. CD22 and CD72 were mainly expressed on CD19+ B cells. The numbers of CD22+ and CD72+ B cells and the percentage of CD22+ B cells and CD72+ B cells in B cells were increased in Eos CRSwNP compared with non-Eos CRSwNP and controls. IgA, IgG and IgM were elevated in both Eos CRSwNP and non-Eos CRSwNP, while IgD and IgE were increased in Eos CRSwNP. CD22 mRNA and CD72 mRNA expression level were negatively correlated with IgE, but not IgA, IgG, IgM and IgD in Eos CRSwNP. The percentage of CD22+ B cells and CD72+ B cells in B cells negatively correlate with the total visual analog scale symptom scores and total endoscope scores.

Conclusion
CD22 and CD72 were significantly increased in Eos CRSwNP patients and negatively correlated with IgE level, indicating the down-regulation role of CD22 and CD72 in B cell activation and a suppressive role in the synthesis of IgE in Eos CRSwNP.
Gut and Oral Microbiota, Immunity and Clinical Features in Taiwanese Children with Atopic Dermatitis
Authors: Yen Hung-Rong , Yang Pei-Shan , Wei Chang-Ching , Wu Po-Yuan 
Published: January 2023

Abstract
Atopic dermatitis (AD) is a skin inflammatory disease, characterized by itch, redness, swelling, oozing, thickening or dryness of skin. The condition typically starts from infancy with changing clinical severity in adolescence and adulthood. It bothers not only the health but also quality of life. The interactions between skin microbes and the skin conditions have been implicated in atopic dermatitis. However, the interplay between host immunity, gut and oral microbiota and clinical features in children remains unclear. In this study, we recruited 30 subjects (17 boys and 13 girls) with atopic dermatitis in a medical center in Taiwan. The mean age was 7.11±5.20 years old. The presentation of skin lesions included redness (73.3%), itch (66.7%), dryness (56.7%) and oozing (36.7%). Some of the patients have comorbidities of allergic rhinitis (30%) and asthma (10%). The mean SCORAD index was 55.33±16.96, with an involvement of 52.58±19.35% of body surface area. The mean eosinophil count was 12.01% and the total IgE was 5040.17 IU/mL. The gut and oral microbiomes were analyzed by using 16S rRNA gene and metagenome sequence analyses. Sterptococcal species are the major oral species and Bacteroides species are the major gut species associated with AD in children. Our data suggest that oral and gut microbiota may underlie the clinical features of AD. Further investigations on the interplay of microbiota, immunity and clinical features are warranted.
Live or let die: Pyroptosis and inflammasome activation in glial cells during multiple sclerosis and experimental autoimmune encephalomyelitis
Authors: McKenzie Brienne , Mamik Manmeet , Saito Leina , Boghozian Roobina , Monaco Maria Chiara , Major Gene , Lu Jian-Qiang , Branton William , Power Christopher 
Published: January 2023

Abstract
Inflammasome-associated caspases mediate the maturation and release of the proinflammatory cytokines IL-1β and IL-18, and activate the pore-forming protein gasdermin-D (GSDMD). Recently, GSDMD was shown to be the primary executioner of pyroptosis, a lytic form of programmed cell death. Excessive GSDMD pore formation causes local osmotic swelling and eventual membrane lysis. Although evidence has emerged for inflammasome activation and pyroptosis in neurological diseases, the role of GSDMD in neuroinflammation remains uncharacterized. Multiple sclerosis (MS) is the prototypic inflammatory demyelinating disease of the central nervous system (CNS). We provide molecular evidence for GSDMD-mediated inflammasome activation and pyroptosis in both macrophages/microglia and, unexpectedly, in myelin-forming oligodendrocytes (ODCs) in the CNS of patients with MS and in the associated animal model, experimental autoimmune encephalomyelitis (EAE). We observe inflammasome activation, GSDMD expression, and pyroptosis in human microglia and ODCs in vitro inflammatory stimulation. Further, we demonstrate that both caspase-1 inhibition by the small molecule inhibitor VX-765 and GSDMD inhibition via siRNAs suppress pyroptosis in human microglia. VX-765 treatment of EAE animals reduced expression of inflammasome- and pyroptosis-associated proteins in the CNS, reduced neuroinflammation, prevented axonal injury, and improved neurobehavioral performance. Thus, GSDMD-mediated pyroptosis in multiple glial populations represents a previously unrecognized mechanism of inflammatory demyelination and represents a unique therapeutic opportunity for mitigating neuroinflammation.
IRF5 hyper-activation is a driver of systemic lupus erythematosus (SLE) onset and severity
Authors: Barnes Betsy J. , De Saurav , Song Su , Nelson Victoria 
Published: January 2023

Abstract
The transcription factor interferon regulatory factor 5 (IRF5) is a central mediator of innate and adaptive immunity. Genetic variations within IRF5 associate with risk for systemic lupus erythematosus (SLE) and mice lacking Irf5 are protected from lupus onset and severity. Here we show that IRF5 genetic variants confer risk by increasing basal IRF5 activation in plasmacytoid dendritic cells resulting in elevated IFNα production. IRF5 hyperactivation in SLE immune cells correlates with SLEDAI and dsDNA titers. Upon characterization of immune cells from NZB/W F1 lupus-prone mice, we found that Irf5 was hyperactivated at 19 weeks-old, which precedes clinical onset. To test whether IRF5 hyperactivation is a driver of SLE, we developed novel therapeutics targeting IRF5 activation. Inhibitors are cell permeable, non-toxic and selectively bind to full-length inactive IRF5 monomer. In vivo analysis revealed functional mimicry with Irf5 knockout mice and treatment of NZB/W F1 mice for a two-week period prior to clinical onset gave significantly reduced dsDNA titers, anti-nuclear antibody (ANA) staining, and improved survival. The IRF5 inhibitor also blocked ex vivo SLE serum-induced IRF5 activation and reversed IRF5 hyperactivation detected in SLE immune cells. Results implicate IRF5 hyperactivation as a risk factor that drives SLE and provide the first pre-clinical support for treating SLE patients or those at risk of SLE with IRF5 inhibitors.
MCP-1 plays a critical role in neutrophil function and pyroptosis during Carbapenemf-Resistant Klebsiella Pneumoniae
Authors: Jin Liliang , Ghimire Laxman , Paudel Sagar , Cai Shanshan , Rangasamy Tirumalai , Jeyaseelan Samithamby 
Published: January 2023

Abstract
Monocyte chemoattractant protein-1 (MCP-1) is important for monocyte recruitment to the lungs in response to bacterial infection. MCP-1 is also essential for protective neutrophil recruitment to the lungs during Escherichia coli pneumoniae infection. Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection has been rapidly growing as a life-threatening nosocomial disease in many countries. Little is known regarding the role of chemokines and early cellular immune responses in protective immunity to pulmonary infection with CRKP. In the current study, we investigated the role of MCP-1 in pulmonary innate immunity against CRKP infection in C57Bl/6 mice and MCP-1−/− mice. MCP-1 appears to be essential for restricting bacterial outgrowth and death in mice. Compared to the C57Bl/6 mice, MCP-1−/− mice showed reduced influx of neutrophils in the airways and lung parenchyma, as assessed by nucleated cell concentrations in BALF and myeloperoxidase activity (MPO) in lung tissue. Moreover, the neutrophil extracellular traps (NETs) induced by CRKP were reduced in MCP-1−/− mice in both in vitro and in vivo experiments. Intriguingly, subsequent to CRKP infection, MCP-1−/− mice demonstrated increased pyroptosis through higher expression of IL-1β and cleaved gasdermin-d in the lung. These findings enhance our understanding of the critical role of MCP-1 in modulating neutrophil function and pyroptosis in neutrophils and suggest that therapies targeting modulation of NETs and pyroptosis during CRKP infection should be explored.
Combining Drop Array Laminar Flow Processing and Image-based Flow Cytometry to Evaluate Changes in Monocyte Intracellular Cytokine Production
Authors: Gary Melody A. , Tanner Elizabeth A. , Stelly Steven , Davis Asheal , Bowman Erin M. , McFarlin Brian Keith 
Published: January 2023

Abstract
The use of image-based flow cytometry has exponentially expanded the knowledge that can be derived regarding cellular changes compared to traditional PMT-based flow cytometry. Other recent advances in cell processing (i.e. Drop Array laminar flow processing) have further increased robustness by protecting cells from loss and damage during flow cytometry processing and labelling. Despite the advances in these respective fields to our knowledge, no study has attempted to combine both technologies to further increase sensitivity and throughput. Thus, the purpose of this report was to compare and evaluate a traditional cell prep workflow to a Drop Array workflow in the preparation of cells for image-based flow cytometry analysis. In order to complete this testing, we selected monocyte LPS-stimulated intracellular monocyte cytokine expression (i.e. IL-6 and TNF-alpha). Monocytes are an ideal cellular target to study because they are commonly lost during traditional centrifuge processing methods. Blood samples were collected from subjects according to IRB approved procedures, the latest Declaration of Helsinki, and a signed consent form. We found that processing with the Drop Array workflow resulted in reduced monocyte loss during staining, reduced cellular debris, and improved noise-to-signal ratio for intracellular IL-6 and TNF-alpha. More research is needed to evaluate the Drop Array workflow for other cell types, but it appears to be an effective technology to processing monocytes for image-based flow cytometry analysis.
Advanced age does not diminish phagocytic activity or NET production in zebrafish (Danio rerio) kidney leukocytes
Authors: Hannum Lynn G. , Chen Tianzeng 
Published: January 2023

Abstract
The pace of growth in populations over the age of 65 is increasing in the U.S. and worldwide, posing serious challenges for health care systems. Studies in both humans and mice have revealed evidence of dysregulation of innate immunity in older individuals, although experimental systems and results of those studies vary widely. To explore whether the zebrafish (Danio rerio) model could be valuable in understanding the effects of aging on innate immune processes, we assessed phagocytic activity, respiratory burst response and neutrophil extracellular trap formation (NETosis) by anterior kidney phagocytes from healthy young (7–8 months old) and aged (&gt;2.5 year old) zebrafish. Exposing leukocytes from transgenic mpx:GFP fish to opsonized pHrodo-labeled killed E. coli for 30 or 60 minutes revealed is no difference in the mean percentage of neutrophils that phagocytosed bacteria between the two ages at either time point. Interestingly, older fish displayed a wider range of responses and a higher level of individual variability than the younger population. Active neutrophils from the aged fish also exhibited a higher mean fluorescence intensity (MFI) than young fish at 30 min incubation, but fluorescence levels were similar in the two populations after 60 minutes of incubation. Leukocytes from older and younger wild type AB fish showed identical respiratory burst responses to PMA. When NET production was stimulated by calcium ionophore, neutrophils from aged AB zebrafish exhibited a higher NET index than the younger cohort. These initial results indicate that older zebrafish preserve a similar level of activity, if not hints of higher activity, in tested neutrophil functions compared to younger fish.