bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mapping the cellular landscape of the maternal-fetal interface in women with preterm birth and chronic chorioamnionitis
Authors: Levenson Dustyn , Xu Yi , Garcia-Flores Valeria , Galaz Jose , Neshek Barbara , Romero Roberto , Gomez-Lopez Nardhy 
Published: November 2024

Abstract
Preterm birth (PTB) affects over 15 million infants yearly. In up to 40% of late PTBs, placental examination reveals chronic chorioamnionitis (CCA), maternal mononuclear cell infiltration in the chorioamniotic membranes; yet, the cellular landscape remains fully unknown. Herein, we first used imaging mass cytometry (IMC) to map the phenotype and spatial distribution of immune cells at the maternal-fetal interface of women with PTB and CCA and non-CCA controls (n = 4 each). Cases exhibited a 5-fold increase in T cells, including effector memory CD4+ (p=0.02) and proliferative CD8+ T cells (p=0.04), and a 7-fold rise in CD163+CD206+ macrophages (p=0.05) compared to controls. NK cells were also present in both groups. Neighborhood analysis revealed that CD8+ T cells are in proximity to macrophages and CD4+Tbet+ T cells, and distanced from NK cells. Next, we performed spatial transcriptomics (STx) in each study group (n = 8 PTB with CCA, n = 7 PTB no CCA). STx highlighted diversity in several clusters of T cells, macrophages, and other immune cells found to be transcriptionally active in CCA. Gene ontology analysis indicated enrichment for CD8+ T cell activation, as well as IL-12 and IFNγ signaling in macrophages in CCA. Ongoing efforts are focused on validation and functional testing of such immune cells in order to unveil the complex interactions at the maternal-fetal interface in CCA, a placenta pathology associated with adverse outcomes in the mother and offspring.
Fungal Chitin Does Not Contribute To Allergic Asthma Severity
Authors: Ellis Diandra , Makullah Mgayya , Jones Mary Jane , Cheung Suki , Willems Marjoleine , Steele Chad 
Published: November 2024

Abstract
Chitin, a polysaccharide found in the fungal cell wall and exoskeletons of house dust mites and cockroaches, has garnered attention as a potential immunoreactive allergen. Mammals have evolved to express chitin-degrading chitinases that may modulate immune responses to chitin. We have previously reported that mice deficient in acidic mammalian chitinase (AMCase, Chia) demonstrated better lung function (i.e. airway hyperresponsiveness, AHR) during allergic fungal asthma. Here, we show that during allergic fungal asthma, mice overexpressing AMCase (SPAM mice) had worse lung function, and chitin-bearing conidia from Aspergillus fumigatus are detectable in the lung. The lung function of mutant mice directly correlated with the level of chitinase activity during allergic fungal asthma (Chia-/- mice, low chitinase activity, lower AHR; SPAM mice, high chitinase activity, higher AHR), suggesting that chitin catabolism promoted AHR. Chronic exposure of normal mice to purified chitin from A. fumigatus resulted in moderate inflammatory changes in the lung, however this was not sufficient to induce AHR. Moreover, chronic exposure of mutant mice to purified chitin did not induce AHR. Collectively, these results indicate that 1) fungal chitin alone is incapable of driving AHR and 2) chitinase-mediated degradation of A. fumigatus chitin may facilitate unmasking and/or liberation of other fungal cell wall components that preferentially drive inflammatory responses which contribute to AHR.
Teaching Immunotherapy at College of Pharmacy-Therapy-based Learning
Authors: Fazal Nadeem , Graefen Bahar 
Published: November 2024

Abstract
Different types of immunotherapies employed as chemotherapy work in different ways. Some immunotherapy treatments help the immune system stop or slow the growth of cancer cells. Immunology course for Pharmacy year 2 students is taught at College of Pharmacy Chicago State University. The perquisites for the course were Medical Physiology, Biochemistry and Pathophysiology. The students had no prior knowledge of basic immunology. The way the course was designed was that they learn about the current available immunotherapy and then were taken back to the basic concepts of immunology to make sense of that module. We divided Pharmacy Immunology course into five modules, i.e., 1) Monoclonal antibodies and immune checkpoint inhibitors, 2) Non-specific immunotherapies, 3) Oncolytic virus therapy, 4) T-cell therapy, 5) Cancer vaccines. Although the students were taught immunotherapies in two-hour long workshops as discussion in teams, they were also tested for their conceptual knowledge of basic immunology. The assessment results consistently showed a higher level of learning according to Bloom’s taxonomy. The exams were designed to test critical thinking and application and synthesis of knowledge of the students. The assessment data suggested that student performance was significantly improved as compared to previous years of Pharmacy year 2 students who were taught similar content without using this teaching methodology.
A novel mouse model of silicosis and silica-induced inflammatory arthritis.
Authors: de Ocampo Caroline , Mayeux Jessica , Kono Dwight , Pollard Michael , Janssen Lisa 
Published: November 2024

Abstract
Introduction: A strong association between silica dust exposure and development of systemic autoimmune diseases, including rheumatoid arthritis (RA), is widely recognized; however, a direct mechanism has yet to be established. To elucidate this connection, we investigated the role of pulmonary silica exposure in promoting silicosis and RA in BXD2/TyJ mice.
Methods: BXD2/TyJ mice exposed to 5 mg crystalline silica or PBS and examined 1-, 2-, 6-, 12-, 20-weeks later. Total immunoglobulin-G and -M, as well as autoantibody levels, measured in serum and bronchioalveolar lavage fluid (BALF), and lung and ankle tissue assessed histologically. Immune cells characterized in lung, spleen, and tracheobronchial lymph node with flow cytometry. B and T cell-containing pulmonary lymphoid accumulations visualized with immunofluorescent staining.
Results: Severe silicosis was observed, and B and T cell-containing lymphoid accumulations were detected in the lung as early as 2-weeks following silica exposure. Markedly increased levels of a range of autoantibodies were detected beginning 2-weeks after silica exposure in BALF and 6-weeks in serum. Significantly higher ankle synovitis and bone erosion was observed beginning at 12-weeks post silica exposure compared to controls.
Conclusion: Silica exposure accelerates severe inflammatory arthritis in a susceptible background, establishing a model to study the connection between mucosal inflammation and rheumatoid arthritis.
Circadian clock disruption exacerbates noncanonical inflammasome-dependent pyroptosis via Rev-erbα-SAA1 axis
Authors: Choi Bokeum , Shim Do-Wan , Eo Jun-Cheol , Kim Saeyoung , Yu Je-Wook 
Published: November 2024

Abstract
Circadian arrhythmia has been linked to the increased susceptibility of multiple inflammatory diseases. However, it remains elusive how disruption of circadian clock modulates innate immune responses including inflammasome signaling. Here, we examined a potential role of circadian clock on inflammasome-mediated responses through myeloid-specific deletion of BMAL1, a master circadian clock regulator. Intriguingly, Bmal1 deficiency significantly enhanced macrophage pyroptosis and lethality of mice under noncanonical inflammasome-activating conditions, but did not alter canonical inflammasome responses. Transcriptome analysis revealed that Bmal1 deficiency led to a marked reduction in Rev-erbα expression at steady state and a significant increase in serum amyloid A1 (SAA1) expression upon poly(I:C) stimulation. We found that Rev-erbα is critical for poly(I:C)-induced SAA1 production, resulting in the circadian oscillation pattern of SAA1 expression in myeloid cells. Mechanistically, Rev-erbα inhibited poly(I:C)-induced association of C/EBPβ to the promoter region of Saa1 leading to the reduced transcription of Saa1 in macrophages. Bmal1-deficient macrophages exhibited enhanced binding of C/EBPβ to Saa1. Collectively, our data demonstrate a potent suppressive role of circadian clock BMAL1 on noncanonical inflammasome response via Rev-erbα-C/EBPβ-SAA1 axis and further suggest that circadian arrhythmia can enhance the susceptibility to noncanonical inflammasome-mediated toxicity.
Building a neural network model to define DNA sequence specificity in V(D)J recombination
Authors: Harris Justin , Fagg Andrew , Wu Dee , Rodgers Karla 
Published: November 2024

Abstract
The initial formation of antigen receptor repertoires occurs in developing B and T cells through V(D)J recombination, which results in the joining of two-to-three gene segments that encode for the antigen binding sites of immunoglobulin and T cell receptors. For a gene segment to be targeted for recombination it must be flanked by a semi-conserved recombination signal sequence (RSS). During each V(D)J recombination event, the RAG1/2 recombinase creates DNA double stranded breaks at two RSS substrates that are then repaired to yield the recombined receptor gene. Gene segments are utilized nonequally in generation of the repertoires. The local regulatory and chromatin environment surrounding gene segments and their flanking RSSs have helped to partially explain the nonuniform use of RSSs across each antigen receptor locus. However, each of these loci have distinctive environments, complicating the generalization of RSS usage in V(D)J recombination across loci. Using recombination data of partially randomized 12RSSs, obtained in a chromatin-free environment, a neural network (NN) was built that delineates how changes in sequence at certain positions in the RSS can affect recombination level. External validation revealed the specific effects of distal regions of the RSS on recombination level. Our NN model demonstrates key principles of the entire RSS sequence in defining gene segment utilization in V(D)J recombination.
Lung Inflammation and Pathology are Mast Cell Dependent in a Model of Formaldehyde Toxicity.
Authors: Gibb Matthew , Reinert Angela , Bauer Alison , Brown Jared 
Published: November 2024

Abstract
The mucosal surface of the respiratory tract is continually exposed to the external environment, including potential toxicants. Mast cells are located throughout the lung and are capable of rapid activation, degranulation, and cytokine release, leading to allergic inflammation and disease. Formaldehyde, a ubiquitous chemical, activates mast cells contributing to airway hyperresponsiveness with subsequent neutrophil recruitment and activation, but little is known about the contribution of mast cells following acute, high dose exposure to formaldehyde. To assess the role of mast cells to acute formaldehyde exposure, we utilized wild type (C57BL/6) and B6.Cg-KitW-sh/HNihr-JaeBsm (Kitw-sh, mast cell deficient) male mice. C57BL/6 mice exhibited dose-dependent decreases in migratory dendritic cells, and statistically significant increases in alveolar macrophages and mast cells as assessed by flow cytometric analysis of lung tissue. Bronchoalveolar lavage analyzed lipid mediators, inflammatory cytokines, and cell counts with a significant and dose-dependent increase in cell counts in C57BL/6 mice where Kitw-sh mice showed no significant increase, regardless of dose. Lung pathology revealed increases in immune cell populations and alveolar septal wall thickening in C57BL/6 mice not seen in Kitw-sh mice. Results indicate mast cells may play a significant role in cell recruitment, inflammation, and pathology in the lung post-exposure to chemical pollutants such as formaldehyde.
HBV immune checkpoint receptor CD33 (Siglec-3) is a promising therapeutic target for chronic hepatitis B infection
Authors: Sung Pei-Shan , Lee Chieh-Ju , Huang Yi-Hsiang , Wu Chang-Ru , Hsieh Shie Liang 
Published: November 2024

Abstract
Approximately 300 million people globally suffer from chronic hepatitis B (CHB), leading to complications like hepatic fibrosis, cirrhosis, and hepatocellular carcinoma. Despite a significant reduction in new HBV infections through vaccination, a definitive CHB cure remains elusive. Current treatments rely on antiviral nucleos(t)ide analogues, often failing to induce crucial anti-HBsAg antibody (HBsAb) production. Our prior study unveiled myeloid CD33 (Siglec-3) as an immune checkpoint receptor for HBV, contributing to HBV-induced tolerance. Blocking CD33 with anti-hCD33 mAb enhances antigen-presenting molecule expression and activation markers in dendritic cells, restoring TLR7 agonist-induced cytokine production in CHB patient PBMCs. We recently identified a novel CD19+ B cell population expressing abundant CD33 in CHB patient PBMCs (5-20% of CD19+ B cells), contrasting with rare detection in healthy donors (0.5-3%). Anti-hCD33 mAb treatment induces HBsAb production in PBMCs and CD19+ B cells. Furthermore, establishing a CHB mouse model via pAAV-HBV-1.2 hydrodynamic injection in human CD33 transgenic mice (hCD33 Tg) demonstrated prolonged HBsAg persistence compared to wild-type mice, suggesting human CD33 impedes HBV clearance. Remarkably, anti-hCD33 mAb administration accelerates HBsAg clearance in hCD33 Tg mice, highlighting CD33 as a promising CHB therapeutic target, with anti-hCD33 mAb potential to reactivate host immunity for HBV clearance in CHB patients.
Multi-modal single-cell sequencing of circulating mononuclear cells deconvolutes outcome-specific immune responses in trauma patients
Authors: Chen Tianmeng , Hughes Julia , Gregory Alyssa , Cornoy Julia , Chen Wei , Duerr Richard , Billiar Timothy 
Published: November 2024

Abstract
Immune dysfunction plays a key role in contributing to high morbidity and mortality after trauma. To identify the immune cell subsets associated with a specific clinical trajectory (SR vs. FR, slow vs. fast recovery), we isolated the peripheral blood mononuclear cells from 16 trauma patients, including 8 SR patients (ICU days &gt; 7) and 8 FR patients (ICU days &lt; 7) at 72 hours after injury, along with 4 age and sex matched healthy controls. A total of 20 samples were subjected to DOGMA-seq, a tri-modal single-cell assay that measures gene expression, open chromatin accessibility, and surface protein expression simultaneously in the same cells. Integrated analysis revealed patient group-specific changes in T cells and monocytes. SR patients had lower numbers of cytotoxic T cells (CTLs) and these remaining CTLs were less cytotoxic. We identified a subset of monocytes overrepresented in FR patients. We extracted the genes specifically upregulated in this subset and queried a published whole blood transcriptome dataset of 167 trauma patients. These genes were positively correlated with T cell activation pathways, and were also correlated with a quicker recovery in this bulk dataset. Thus, we were able to validate our findings derived from DOGMA-seq from 16 trauma patients in a much larger cohort. Our analysis provides a landscape of immune responses in trauma patients based on clinical trajectories and characterizes the outcome-specific changes across cell types.
USP30 inhibition augments mitophagy to prevent T cell exhaustion.
Authors: Xin Gang , Zhang Ruohan , Li Jianying , Yang Mingchong , Jin Jiacheng , Pich Thera , Sun Nuo , Xin Gang 
Published: November 2024

Abstract
The quality control of mitochondria through mitophagy is crucial for maintaining effective anti-tumor CD8 T cell responses. However, this process is compromised during T cell exhaustion. The specific mechanisms regulating mitophagy remain unclear, hindering efforts to target it for reversing T cell dysfunction. To tackle this challenge, we employed a mitophagy-reporter mouse model and revealed impaired mitophagy activity in exhausted CD8 T cells exhibiting mitochondrial dysfunction. Through a bioinformatic and experimental approach, we identified that USP30, a mitochondrial deubiquitinase inhibiting mitophagy, is upregulated in exhausted CD8 T cells with the decline of mitophagy. Genetic deletion or pharmacological inhibition of USP30 can significantly enhance mitophagy in exhausted CD8 T cells and restore their mitochondrial fitness and effector function, ultimately reinvigorating anti-tumor CD8 T cell responses and enhancing immunity against tumors. Furthermore, our observations indicated that the stimulation of the T-cell receptor amplifies the transcription of USP30 in exhausted CD8 T cells, highlighting its pivotal role in regulating mitophagy. Overall, our findings shed light on the vital role of USP30-regulated mitophagy in controlling mitochondrial quality during T cell exhaustion while also providing valuable insights into developing targeted therapies to restore mitophagy, thereby improving the immune response against tumors.
Opposing roles of STAT1 and STAT3 in COX-2 protein expression in human follicular dendritic cells
Authors: Jeong Jihye , Choe Jongseon 
Published: November 2024

Abstract
Prostaglandins (PGs) are the lipid mediators of inflammatory responses and acknowledged also as the mediators of immune inflammation. We have explored the role of PGs and their production mechanisms using follicular dendritic cell (FDC)-like cells. In the course of our recent study of the regulation mechanisms of PG production, we observed the involvement of JAK-STAT molecules in the IL-1β-induced PG production. In the current study, we aimed to elucidate the role of JAK and STAT proteins in the expression of COX-2 protein, the responsible enzyme for PG production. COX-2 protein level and PG production were assessed using immunoblotting and EIA, respectively. The intracellular target proteins were detected by a confocal microscopy. STAT1 knockdown resulted in an increase of COX-2 expression, while STAT3 knockdown led to a reduction in the COX-2 induction by IL-1β. IL-1β induced phosphorylation of both STAT1 and STAT3 at the S727 sites, rather than at Y701 or Y705. The phosphorylation and nuclear translocation of pSTAT1 and pSTAT3 were confirmed by a confocal microscopy. Based on these findings, we maintain that STAT1 may play an inhibitory role in COX-2 induction, while STAT3 acts as a promoter, via fine tuning of phosphorylation levels. The current study may enhance our understanding of the precise roles of these STAT proteins in COX-2 induction and PG production and suggests that these molecules could be potential targets for the treatment of immune inflammatory disorders.
A novel interaction between complement product C4a and the receptor for advanced glycation end products (RAGE)
Authors: Wang Fang , Wu Yee 
Published: November 2024

Abstract
Complement proteins (C) including their split products play important roles in host defense and homeostasis. Anaphylatoxins C3a and C5a, by binding to their receptors on immune and non-immune cells, mediate cell recruitment and the release of proinflammatory molecules. The receptor and function of C4a remain unclear. C3a, however, in addition to its conventional receptor C3aR, is known to bind to the receptor for advanced glycation end products (RAGE), a receptor implicated in initiating and sustaining inflammatory responses. This interaction can also be mediated by the TLR9 agonist, CpG. C4a is similar to C3a in its structure and net charge. Therefore, we hypothesize that C4a may interact with RAGE. We performed co-immunoprecipitation experiment on 293T cell lysate expressing C4a and RAGE and found that RAGE did not co-immunoprecipitate with C4a, indicating that C4a does not directly interact with RAGE. Using fluorescently labelled C4a protein, we found that there was no significant C4a binding to 293T cells irrespective of RAGE overexpression by flow cytometry. However, by preincubating C4a with CpG, we detected elevated C4a binding to RAGE-expressing 293T cells compared to control cells. These findings suggest that CpG mediates interaction between C4a and RAGE. Our ongoing investigations focus on determining the binding properties and the functional impact on inflammatory response of this novel interaction.
Obesity reprograms macrophage nucleotide metabolism to enable hyperinflammation
Authors: LIU DANHUI , Liang Shuang , Zhong Zhenyu 
Published: November 2024

Abstract
Obesity-induced NLRP3 inflammasome hyperactivation is a major risk factor for human diseases spanning autoimmune, degenerative, metabolic disorders, and cancer. However, its underlying mechanisms remain elusive. Here we show that obesity compromises the function of SAMHD1 – an evolutionarily-conserved deoxyribonucleotide triphosphate (dNTP) hydrolase essential for cellular nucleotide homeostasis. This dysregulation leads to aberrant accumulation of cytosolic dNTPs in macrophages, which are subsequently transported into mitochondria, and directly serve as building blocks for new mitochondrial DNA (mtDNA) synthesis bypassing the CMPK2-mediated nucleotide salvage synthesis pathway. The uncontrolled mtDNA neosythesis then triggers the overproduction of oxidized mtDNA upon NLRP3 activators stimulation, ultimately resulting in hyperactivation of NLRP3 inflammasome. Genetic ablation of mouse Samhd1 leads to overwhelmed NLRP3 inflammasome activation in vivo and exacerbates immunopathology in multiple NLRP3-dependent disease models. Moreover, mice lacking SAMHD1 exhibit heightened circulating IL-1β, insulin resistance, and metabolic dysfunction-associated steatohepatitis (MASH) upon high-fat diet feeding. Overall, our study establishes SAMHD1 as a macrophage-intrinsic valve restraining NLRP3 inflammasome activation, illuminating that obesity, through compromising SAMHD1's function, rewires macrophage nucleotide metabolism to enable hyperinflammation and disease development.
Emergence of novel CD3+ Macrophages in Brown Adipose Tissue Following Adrenergic Stimulation
Authors: Chae Hee-Don , Levi Jelena 
Published: November 2024

Abstract
Macrophages play a vital role in maintaining tissue homeostasis and defending against infections and cancer. Brown adipose tissue (BAT) is essential for expending surplus energy during cold stress, while white adipose tissue functions as the primary energy reservoir. The sympathetic nervous system responds to cold stress by releasing norepinephrine (NE), activating BAT through β3-adrenergic receptor. BAT-infiltrated macrophages exert opposing effects on thermogenesis, regulating the interplay between BAT cells and sympathetic nerves to either inhibit thermogenesis by clearing NE or enhance it by supporting sympathetic innervation of BAT.
In mouse interscapular BAT, we identified a novel subset of CD3+ macrophages (CD45+ CD3+ CD4+ CD8+ CD11b+ F4/80+), whose frequency increased with chronic adrenergic stimulation. Despite the potential for T cell-monocyte doublets in flow cytometry, CD3+ macrophages demonstrated a unique surface expression pattern, lacking T cell markers CD62L and TCRβ. Furthermore, they showed elevated levels of Ly6C, MHCII, F4/80, and CD11b compared to T cells and CD3- macrophages. CD3+ macrophages exhibit heterogeneity in MHCII expression, with subsets expressing low and high levels.
Since MHCII-expressing macrophages play a crucial role in regulating sympathetic nerve-BAT interaction, further characterization of CD3+ macrophages is pivotal for understanding the intricate mechanisms that govern adaptive thermogenesis regulation.