bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intricate mechanisms trigger scattered crypt IEC apoptosis mediated necrotizing enterocolitis pathogenesis in neonatal mice
Authors: Subramanian Saravanan , Bu Heng-Fu , Chou Pauline , Wang Xiao , De Plaen Isabelle , Tan Xiao-Di 
Published: November 2024

Abstract
Necrotizing enterocolitis (NEC) is a life-threatening gastrointestinal inflammatory disorder among premature infants. Intestinal epithelial cell (IEC) apoptosis contributes to NEC pathogenesis. However, how scattered crypt IEC apoptosis leads to NEC with excessive epithelial necrosis in intestinal villi and mucosa inflammation remain unclear. We developed a novel triple-transgenic mouse model, namely, 3xTg-iAPcIEC (inducible apoptosis phenotype in crypt-IEC) using doxycycline (Dox)-inducible tetO-rtTA system and vil-cre technology, for inducing tissue-specific overexpression of Fasl in IECs. We found that scattered crypt IEC apoptosis caused intestinal crypt inflammation and villous necrosis resembling NEC in neonates. This pathological progression initiated a set of NEC-associated intricate cellular and molecular pathophysiological effect including increase in Rip3 and Ifng, infiltration of CD8+ T cells, and dysbiosis with Gram-positive bacteria. Mechanistically, scattered crypt IEC apoptosis-induced IFN-γ and RIP3 activation were noted to mediate both intestinal crypt inflammation and mucosal villous necrosis, whereas CD8+ T cells and dysbiosis with Gram-positive bacteria contribute to mucosal villous necrosis. Notably, we observed that blocking any of these events protects NEC development in 3xTg-iAPcIEC mouse pups, underlining their central roles in the NEC pathogenesis. Our findings may advance knowledge in preventing or treating this devastating disorder in humans.
Case report: Use of emapalumab-lzsg for secondary hemophagocytic lymphohistiocytosis in a patient with chronic granulomatous disease
Authors: Patel Raveena , Gunderson Carly , Ishikawa Caren , Alqahtani Mashael , Cadilla Adriana , Douglas Dorothea 
Published: November 2024

Abstract
Background: Chronic granulomatous disease (CGD) is an immunodeficiency marked by infections due to NADPH oxidase defects. A hyperinflammatory response predisposes to granuloma formation and hemophagocytic lymphohistiocytosis (HLH), a syndrome defined by uncontrolled cytokine release. We present an adolescent with undiagnosed CGD who developed secondary HLH, requiring the use of emapalumab-lzsg, a human anti-interferon-gamma antibody. To our knowledge, this is the first report of emapalumab-Izsg use in a patient with secondary HLH with CGD.
Case Presentation: A 16-year-old male with Chron’s disease presented with six months of fevers. Imaging showed reticulonodular disease and granulomas throughout the lungs. Respiratory status deteriorated, requiring intubation and ECMO. Blood cultures grew Burkholderia cepacia. Flow cytometry was abnormal and genetic tests confirmed CGD. Further clinical and lab derangements led to HLH testing. HLH was confirmed and treated with steroids, etoposide, and intravenous immunoglobulin. With no clinical improvement, emapalumab-Izsg was recommended. Six-week course of emapalumab-Izsg led to disease regression, with decline of CXCL-9, a marker of active disease.
Conclusion: Emapalumab-Izsg has been used in primary HLH while utility in secondary HLH remains undefined. The standard of care for secondary HLH includes immunosuppressive and chemotherapeutic therapies. We propose that emapalumab-Izsg be strongly considered an option in secondary HLH.
Alveolar macrophage (AM) Toll-like Receptors (TLRs) signaling drives inflammation and pathogenic neutrophilic infiltrate during neonatal murine Influenza virus infection
Authors: Rao Abhishek , Kumova Ogan , Onufer Abigail , Pascasio Judy , Carey Alison 
Published: November 2024

Abstract
Like human neonates, murine neonates are highly susceptible to influenza viral (IV) infection. AMs are tissue-resident immune cells which are activated through TLRs to initiate immune cell recruitment to the site of infection, critical for viral control. However, bystander damage can also occur in the delicate alveoli. Therefore, we hypothesized that AM TLRs contribute to neonatal murine IV pathogenesis via neutrophil recruitment. To determine the relative contribution of AM TLR signaling, we utilized the Cre-flox system to generate AM-specific TLR deletions (LysMCre-MyD88flox). Three-day-old transgenic mice were infected with influenza type A virus strain PR/8/34 (H1N1) and tracked for survival. LysMCre-MyD88flox neonates demonstrated improved survival (70%) compared to wild type (WT) (33%). Moreover, LysMCre-MyD88flox neonates had decreased inflammatory cytokine concentrations in the broncho-alveolar lavage (BAL) compared to WT, specifically IL-18, IL-6, TNF-α and myeloid recruiting chemokines MCP-1 and CXCL1. Flow analysis of the BAL revealed lower neutrophils in the LysMCre-MyD88flox group, consistent with the cytokine concentrations. To test the pathogenic role of neutrophils during IV infection, infected WT neonates were neutrophil-depleted and had improved survival (60% vs. 24%, isotype group). These findings suggest AM TLRs increase IV pathogenesis in neonatal mice and that TLR-blockade is a potential therapeutic target in this vulnerable population.
Influenza virus (IV)-induced Type I Interferons Disrupt Alveolar Epithelial Barrier Function in the Developing Neonate
Authors: Onufer Abigail , Chang-Mell Joshua , Cort Laura , Rao Abhishek , Mdluli Nonto , Pascasio Judy , Carey Alison 
Published: November 2024

Abstract
Type I interferons (IFN-I) limit IV replication in adults. However, IV-infected murine neonates lacking a functional IFN-I receptor (IFNAR-/-) have significantly improved survival and reduced lung pathology relative to wild-type (WT) neonates, indicating IFN-I toxicity in neonates. We hypothesized that IV-induced IFN-I signaling in type II alveolar epithelial cells (AT2s), the primary cell type of IV infection and initiator of host response, contribute to neonatal viral pathogenesis. Therefore, we performed RNA-Seq on AT2s purified from IV-infected IFNAR-/- and WT neonates at 1-, 2-, and 3-days post-infection (DPI). Pairwise comparisons highlighted differentially expressed genes (DEGs) dependent on genotype and timepoint, with DEGs highest between infected IFNAR-/- and WT neonates at 2 DPI and significant pathway upregulation of organ repair. Therefore, we hypothesized neonatal IV-infected IFNAR-/- AT2s possess improved barrier integrity early in infection. Indeed, genes associated with barrier dysfunction (il1b, tgfb1) increased in WT neonates over time, while tight junction gene cldn8 is higher in IFNAR-/- neonates. In addition, IF-stained IFNAR-/- lungs have improved expression of tight junction molecules, ZO-1 and occludin, in comparison to WT neonates at 6 DPI, a time of peak mortality. This suggests that early transcriptional differences in neonatal IFNAR-/- AT2s contribute to increased barrier integrity, mitigating later IV-induced damage and lung pathology.
The role of CXCR4-CXCL12 in the development and migration of human natural killer cells
Authors: Eisman Shira , Koenigsberg Batya , van den Haak Frederique , Pedroza Luis , Hegewisch-Solloa Everardo , Shannon Michael , Mace Emily 
Published: November 2024

Abstract
Natural killer (NK) cells play a critical role in controlling viral infection and malignancy and undergo differentiation from multipotent progenitors in secondary lymphoid tissue. Despite dependence on the microenvironment for differentiation, little is known about the cell-cell interactions that promote NK cell precursor trafficking and maturation in tissue. We have identified the chemokine receptor CXCR4 and its ligand CXCL12 as regulators of the cellular interactions between human NK cell precursors and stromal cells that support NK cell differentiation. Using in-vitro NK cell differentiation, we found that treatment of CD34+ precursors with CXCR4 antagonist AMD3100 leads to a lower efficiency of NK cell generation. Since previous work demonstrates the association of cell migration with NK cell development, we interrogated the effect of AMD3100 on NK cell migration. Using live cell imaging we found that AMD3100 treatment of NK cells or progenitor cells results in altered migratory properties and decreased arrest of cells on CXCL12+ stromal cells. Additional experiments using high resolution microscopy and flow cytometry suggest that integrins are interacting with CXCR4 in NK cells. Our data suggests that the CXCR4-CXCL12 interaction is critical for the migration and proper development of NK cells. Future studies will focus on the necessity for CXCR4-CXCL12 in specific key transitions in development and interrogate the cell biological mechanisms of this process.
N-glycan signature of neutrophil function in a Staphylococcus aureus skin infection model by mass spectrometry imaging
Authors: Bergamin de Castro Tialfi , Ntiamoah Abena , Wu Meng-Jen , Archer Nathan , Scott Alison 
Published: November 2024

Abstract
Staphylococcus aureus (Sa) is a public health concern due to the rise of antibiotic-resistant strains. In humans, elevated risk is associated with Tumor Necrosis Factor (TNF) inhibition. Neutrophils expressing TNF receptors (TNFR1 and TNFR2) respond to Sa skin infection in a mouse model. TNFR2 activation induces PAD4 (peptidylarginine-deiminase-4) which leads to neutrophil extracellular traps (NET) synthesis, increasing host defenses. Alterations in PAD4 and N-glycans are related to increased skin infections and the spatial glycosylation signature during the skin response to Sa is unknown. We compared N-glycan signatures in neutrophil region between wildtype (WT), TNFR1 KO, TNFR2 KO and PAD4-/- groups. Mice were intradermally injected with Sa and skin samples collected for analysis by mass spectrometry imaging. We sprayed PNGaseF on prepared FFPE sections to cleave the N-glycans, sprayed with CHCA matrix, and evaluated in positive ion mode at 50mm spatial resolution on a timsTOF Flex (Bruker Daltonics). Differential N-glycan signatures were identified using a histology-guided approach limiting the comparisons to the PMN region. PAD4-/- showed higher intensity of N-glycans than all groups. However, specific N-glycans were higher in TNFR1 KO than PAD4-/- and TNFR2. A lower profile was found in TNFR2 KO vs. WT. We concluded that each group has a specific N-glycan signature which can be attributed to unique immune responses to Sa infection and NET formation.
role of muscle regenerative macrophages in adjuvant-mediated immune response skewing to seasonal influenza vaccine
Authors: Noureddine Moataz , Chang Lauren , Laghlali Gabriel , Warang Prajakta , Prellberg Mathew , De Geest Bruno , Coughlan Lynda , Schotsaert Michael 
Published: November 2024

Abstract
MF59 (AddaVax-like) is a clinically approved nanoemulsion adjuvant with an unclear mechanism of action. Here we introduce a novel approach to investigating the role of muscle regeneration in adjuvanted vaccine response. We found that AddaVax adjuvanted quadrivalent inactivated flu vaccine (QIV) induced a Th2 immune response with IL-4, -5, and -13 expression in the lungs 5 days after H1N1 lethal challenge. In contrast, IMDQ-PC (TLR7/8 agonist) induced a Th1 lung response and higher IgG2a titers. In BMDM cultures, we found AddaVax unlike IMDQ-PC to require IFNγ co-incubation for the induction of iNOS+ proinflammatory M1 phenotype. Since IFNγ is crucial for muscle regeneration, we hypothesized that AddaVax triggers macrophage-dependent muscle regeneration which leads to an enhanced Th2 vaccine response. We found that AddaVax adjuvanted QIV led to an increase of Arg-I+ M2 macrophages in the muscle 4 days post-injection, similar to late-stage Th2 response in regeneration. Using IgG titers, Our hypothesis was further supported by finding AddaVax response to be diminished in aged mice where regeneration is impaired, but rescuable by depletion of circulating monocytes and reconstitution with young BMDMs. Using in-depth characterization and selective depletion, we will investigate the crosstalk between muscle regeneration and AddaVax response allowing a better understanding of the potential muscle role in inhibiting the Th1 response necessary in viral infections, and cancer vaccines.
Hands-on gene recombination as an active learning tool for TCR and BCR assembly
Authors: Tritz Zachariah 
Published: November 2024

Abstract
Active learning approaches within STEM been shown superior to didactic lectures. I queried whether active learning for the difficult concept of B and T cell receptor (B/TCR) recombination promoted material retention in subsequent examinations compared to a lecture. A 2020 publication by Stranford et al. put forward an active learning tool for this topic using pieces of paper to stand in for genes, allowing students to physically recombine these “genes” with scissors and tape to show recombination of BCR and TCRs. I used an adapted version of this activity in an undergraduate immunology course at University of Wisconsin–La Crosse during the Fall 2023 semester. One section of the course (N=34) received a lecture on the topic while an experimental section of the course (N=26) participated in the activity. A questionnaire assessed student enjoyment of the activity and their perception of whether they learned better with an activity than they would have with a lecture. During the following class period, a multiple choice activity was used to determine student understanding of the material. Students that self-reported better understanding of the material with an activity did perform significantly better than their peers (p=0.045), despite the fact that these students do not have a significantly higher overall course grade (p=0.19). This suggests that a hands-on activity can be an effective way to promote student retention of content related to antigen receptor recombination.
Integrating biomedical sciences (Physiology, Biochemistry, and Cell biology) into pharmaceutical sciences curriculum
Authors: Fazal Nadeem , Graefen Bahar 
Published: November 2024

Abstract
Like other healthcare and medical professional curricula, pharmacy is also struggling to incorporate basic medical sciences into pharmaceutical sciences. Enrollment issues complexed with disappearing admission test like (PCAT) have challenged the instructors to make sure that the first- and second-year pharmacy students are thoroughly prepared with foundational science backgrounds before they start learning clinical and experiential subjects. At the College of Health Sciences and Pharmacy Chicago State University, we prepared and tested ourselves for the same challenge. Immunology taught as a core subject in Pharmacy year-2 was one of the subjects chosen for addressing this issue. Immunology is a rapidly developing biomedical science that delves into the wondrous complexities of the immune system; an extraordinary defense mechanism system evolved to safeguard organisms from diverse threats. At the core of immunology, it overlaps basic cell biology and has an intricate network of cells, tissues, and molecules that need to be introduced to pharmacy students when teaching about the basics of the subject. Retrospective data was analyzed where Immunology was bundled with a 4-credit Microbiology courseas compared to a 2-credit Immunology course which was shifted to Pharmacy year 2. The data suggested that student’sperformance was enhanced when Immunology was offered as a standalone course in Pharmacy Year-2.
Cardiac resident macrophage affects doxorubicin-induced cardiomyopathy
Authors: Jeon Sae-Bom , Park Sahng Wook , Lee Seung-Hyun 
Published: November 2024

Abstract
Doxorubicin is an anticancer drug commonly used in patient with lymphoma or breast cancer, and its main side effect is cardiotoxicity by damaging cardiomyocytes. While the mecahnisms of doxorubicin-induced cardiotoxicity are poorly understood. Recent studies demonstrated that the cardiac resident macrophages play essential roles in regulating homeostasis of cardiomyocyte, promoting electrical conduction, and eliminating dead cardiomyocytes and debris. In this study, we showed that cardiac resident macrophages were involved in cardiomyocyte death in a mouse model of doxorubicin-induced cardiotoxicity. Mice were injected with 20 mg/kg of doxorubicin and their hearts were harvested to determine the composition of immune cells in the heart. Using FACS analysis, we observed that the number of CD45+Ly6GintCD11b+F4/80+ macrophages per mg was significantly reduced in the hearts of doxorubicin-injected mice. These cells are cardiac resident macrophages with a CCR2- MHCⅡ+ phenotype. Also, The TUNEL assay confirmed that macrophage death observed in the cardiac tissue of mice 1 day following doxorubicin administration. Finally, we treated iPSC-derived cardiomyocytes and macrophages with doxorubicin and observed cell death, and found that macrophages were more sensitive to doxorubicin than cardiomyocytes. These results suggest that cardiac resident macrophages are sensitive to doxorubicin, which can affect the death of neighboring cardiomyocytes and lead to cardiotoxicity.
Homeostatic Type I IFN Response at the Placenta Contributes to Sex-biased Neurodevelopmental Outcome
Authors: Kim Eunha , Lai Jenny , Sun Yanyi , Kim Minjin , Choi Gloria , Lee Alice , Huh Jun 
Published: November 2024

Abstract
The maternal-fetal interface, placenta, plays pivotal roles in fetal development. Recent evidence shows that the placenta contributes to the sex-biased prevalence of some developmental disorders such as neurodevelopmental disorders. Commencing an exploration into the molecular intricacies of fetal sex-specific transcriptional changes within the placenta, our study leveraged single-cell RNA sequencing. We uncovered a diminished expression of interferon (IFN)-responsive genes (ISGs) in male fetus-associated trophoblasts compared to female, which have pivotal roles in the process of spiral artery remodeling. We further found maternal NK cells associated with male fetus placenta showed a reduction in the interferon production pathway. The introduction of maternal immune activation (MIA), which induces neurodevelopmental abnormalities in male offspring, it reversed these sex-specific disparities in the placenta. In pursuit of unraveling the potential link between sex-specific ISGs expression and neurodevelopmental outcomes in MIA offspring, we selectively removed IFN alpha and beta receptor subunit 1 (IFNAR1) in a trophoblast-specific manner. Blunting IFNAR1 in the placenta protected male offspring from MIA-induced behavioral abnormalities. Our data suggests the intricate landscape of sex-specific IFN response at the maternal-fetal interface and underscore its role in shaping the sex-bias observed in MIA-induced neurodevelopmental phenotypes.
Heightened infiltration of immune effector cell types in the hippocampal regions in the APP/PS1 mouse model of Alzheimer’s disease.
Authors: Maynes Mark , Ayasoufi Katayoun , Wolf Delaney , Tritz Zachariah , Jin Fang , Hansen Michael , Johnson Aaron 
Published: November 2024

Abstract
Alzheimer's disease (AD) is among the most common neurodegenerative diseases in which a role for immune cells in its etiology is only beginning to be realized. AD has two molecular pathological hallmarks: the presence of amyloid beta (Aβ) plaques and tau tangles within neuronal cells. The amyloid plaques can be modeled in mice by inserting transgenes that enhance human Aβ to create plaques inside the mouse brain. Amyloid precursor protein (Mo/HuAPP695swe) and a mutant human presenilin 1 (PS1-dE9) are two transgenes inserted into C57BL/6 mice that results in Aβ plaques in the hippocampus and cortex. This mouse is known as the APP/PS1 mouse. Immune cell infiltration into the brain is associated with aged APP/PS1 mice. However, comprehensive immune profiling has not been performed in this model to outline immune cell effectors potentially involved in neuropathology. Therefore, we employed spectral flow cytometry to investigate the immune cell profile in 24-month-old APP/PS1 mice compared to an age matched control. There are increased numbers of CD8 T, γδ T, and NK cells expressing perforin and other activation related surface markers. Using small animal MRI imaging, we also observed enlarged ventricles mice indicative of brain atrophy in APP/PS1 mice when compared to wildtype age matched control. These results provide insight into the regional immune cell infiltration of the brain, setting the stage for further analysis of immune mechanisms of neuropathology in experimental AD.
Dysfunctional CD8 T cells in tumors are addicted to antigen
Authors: Erwin Megan , Philip Mary 
Published: November 2024

Abstract
Chronic antigen stimulation, as during cancer or chronic infection, leads to T cell dysfunction. We studied the impact of chronic antigen stimulation on T cell receptor (TCR) signaling in CD8 T cells using a genetic liver cancer mouse model. Tumor-specific CD8 T cells (TST) initially acquired proximal TCR signaling defects, but with increased antigen exposure showed both proximal and distal signaling defects. Despite impaired TCR signaling, a stable population of TST was maintained throughout tumorigenesis. We tried to rescue dysfunctional TST by removing them from tumors but found that TST failed to survive in tumor-free mice; thus, T cells are addicted to antigen, though antigen impairs TCR signaling. We investigated the mechanism of antigen-dependent TST persistence and found that throughout tumorigenesis, a subset of terminally-differentiated (TCF1-) TST within liver tumors could be found in cell cycle. The proliferation-associated distal TCR signaling pathways MAPK and mTOR were impaired in TCF1- TST, and we hypothesize that antigen-dependent persistence is NFAT-dependent. Studies are underway to determine whether all TCF1- TST can enter cell cycle in a stochastic, NFAT-driven manner, or if only a specific subset of TCF1- TST proliferate. By understanding how T cell signaling is regulated in response to chronic antigen stimulation, we can design strategies to selectively modulate TCR signaling to maintain proliferation while preventing loss of effector function.
Liver metastases reprogram neutrophils to potentiate cancer progression
Authors: Pearson Ashley , Holcomb Erin , James Jadyn , Wang Zhuwen , Kyi Justine , Huber Amanda , Green Michael 
Published: November 2024

Abstract
Liver metastases are associated with poor overall survival and increased disease progression, regardless of primary tumor type. Liver metastases suppress systemic antitumor immunity to limit immunotherapy efficacy on primary tumors through hepatic siphoning and deletion of CD8+ T cells, induction of T regulatory cells, and depletion of dendritic cells. However, it remains unclear how liver metastases promote the metastatic cascade and subsequent disease progression. To investigate how liver metastases reprogram the immune system to potentiate disease progression, we use a preclinical polymetastatic murine model of liver metastasis that spontaneously progresses to the lungs. Using B16F10 melanoma, 4T1 triple negative breast cancer, and LLC non-small cell lung cancer in syngeneic C57Bl/6 or BALB/c mice, we observed that the presence of liver metastases increased lung metastatic burden. We hypothesized liver metastases potentiate the formation of a tumorigenic, immunosuppressive premetastatic niche in distant sites. We conducted single cell RNA sequencing, flow cytometry, and immunofluorescence of the liver, peripheral blood, and lungs of mice with or without liver metastases. These data suggest an association between Ly6g+Ly6c- neutrophils and progression in mice with liver metastases. Future efforts will define the contribution of neutrophils to disease progression with the ultimate goal of understanding and addressing the immunosuppression promoted by liver metastases.