bluuebunny/crossref_metadata_2025_split huggingface dataset:
Plasmid-encoded CCL27 co-delivery enhances heterologous protection from respiratory virus challenge
Authors: Gary Ebony , Tursi Nicholas , Warner Bryce , Trachtman Abigail , Patel Ami , Kobasa Darwyn , Weiner David 
Published: November 2024

Abstract
Many pathogens enter the host at mucosal sites. Peripheral vaccination supports robust antigen-specific cellular and humoral immunity but it requires active infection to pull antigen-specific memory cells to the mucosa and relies on passive transudation of antibodies to mucosal surfaces. We reported that co-delivery of the mucosal homing chemokine CCL27 (CTACK) with a DNA plasmid encoding parental SARS-CoV-2 spike (pS) generates antigen-specific IgA in the lung, antigen-specific CD8+ T cells at mucosal surfaces, and was 100% protective in a heterologous Delta VOC challenge model. Here, we extend this work and demonstrate that co-delivery of plasmid-encoded CTACK (+pCTACK) with pS supports protection from heterologous challenge with SARS-CoV-Omicron variants. pCTACK co-delivery significantly lowered viral loads when mice were challenged with Omicron variants BA.2 and XBB.1.5. In the context of Influenza HA DNA vaccination, co-delivery of pCTACK increased HA-specific antibodies in bronchoalveolar lavage, increased antigen-specific CD8+T cells in the lung mucosa, and altered challenge outcomes in vivo. These data have broad implications for the generation of mucosal immunity with parenteral vaccination and support the use of CCL27 to augment parenteral vaccine-induced immunity at mucosal surfaces and support improved outcomes following respiratory virus infection.
Uncovering Alterations in the Murine Interferon Response due to dsRNA Mycovirus during Aspergillus fumigatus Infection
Authors: Rapp Alexander , Obar Joshua , Cramer Robert , Wang Xi , Ross Brandon , Olive Andrew , Stajich Jason 
Published: November 2024

Abstract
Aspergillus fumigatus is a mold that causes an array of diseases, the most severe form being invasive aspergillosis which occurs in immunocompromised individuals. We demonstrated that tissue-resident alveolar macrophages are critical for maintaining host resistance against Aspergillus fumigatus due to their role in the early type I/III interferon response. Specifically, CD11c-Cre x Mavsfl/fl mice have a decreased interferon response and survival outcomes when challenged with A. fumigatus, which can be functionally restored via adoptive transfer of wild-type SiglecF+ alveolar macrophages. Why alveolar macrophages are potent inducers of the type I/III interferon response after Aspergillus fumigatus challenge remains elusive. The type I/III interferon responses were partially related to the ability of alveolar macrophages to recognize mycoviral double-stranded RNA (dsRNA). Aspergillus fumigatus strains infected with polymycovirus 1 (PmV1) are shown to induce a robust type I/III interferon responses both in vitro and in vivo. Critically, Mda5/MAVS-dependent interferon response can still be induced in a fungal dsRNA independent manner, which warrants further explorations. In this study we identified key traits of alveolar macrophages which enable robust Mda5/MAVS signaling necessary for initiating the Aspergillus fumigatus induced interferon response, which maintains host resistance against this ubiquitous mold, through both fungal dsRNA-dependent and -independent mechanisms.
Phenotyping the complex human peripheral blood (PB) B cell compartment using multimodal Cellular Indexing of Transcriptomes and Epitopes (CITE-seq) analysis.
Authors: Meinhardt Nathan , Veltri Anthony , Dittel Bonnie 
Published: November 2024

Abstract
Classifying the heterogeneous human B cell compartment into discrete subsets based on surface phenotype alone remains a major challenge in the field of B cell biology. Our lab has discovered a novel mouse IgM+IgDlow/- B cell subset (BDL) that induces Treg proliferation, making it a potentially significant therapeutic target for autoimmune diseases. But due to their rarity and lack of sufficient cell surface identifiers, a more definitive phenotype is required to track this B cell subset in humans. The objective of these studies was to characterize human IgDlow/- B cells from PB using multiomic CITE-seq analysis. Here, we adapted a standardized gating scheme (CD27xIgD → CD24xCD38) that captures all the major canonical human PB B cell subsets. This scheme allowed us to analyze FACS-purified CD19+IgM+IgDlow/- PB B cells using CITE-seq by targeting these four protein markers and recreate biaxial plots similar to our flow cytometry analyses. It is increasingly understood that mRNA levels within a cell do not correlate well with cell surface protein expression. This represents a major drawback to using a transcriptome-only scRNA-seq approach because any ‘novel’ cell populations discovered based on the upregulation of surface protein transcripts may prove impossible to validate. These studies have established a workflow for a multiomic approach to identify discrete human B cell subsets based on cell surface expression that can be analyzed for transcriptional differences.
Sub-second release of Neurotransmitters from CD4+ T lymphocytes
Authors: Seibold Jordan , Ross Ashley 
Published: November 2024

Abstract
The neuroimmune system is complex and is regulated by small chemical messengers released from resident immune and neuronal cells. Catecholamines like dopamine and norepinephrine are important chemical messengers responsible for initiating and propagating messages between neurons and immune cells; however, the mechanism and dynamics of this signaling are relatively unknown. The presence of the intracellular machinery to synthesize and interact with catecholamines in T lymphocytes has been known for over a decade; however, direct measurement of catecholamine secretion in real time from immune cells has not been possible. Here, we have discovered that Naïve CD4+ T cells are capable of releasing catecholamines on a sub-second time scale. We have used a revolutionary technique, often used in the neuroscience field, called fast-scan cyclic voltammetry (FSCV) to measure sub-second fluctuations of catecholamines from isolated CD4+ T cells. This work provides direct evidence that T cells are capable of releasing catecholamines; potentially as a feedback communicator to resident sympathetic neurons in host immune organs. These results also prove that immune cells can signal on the same time scale as neurons. This work will hopefully open the door to enable more sophisticated studies at the neuron-immune synapse in the future and the understanding of neuro-immune based diseases.
Induction of CXCL3 in airway epithelial cells by a novel IL-9 receptor complex
Authors: Cheung Cherry Cheuk Lam , Abdelaal Ahmed , Kaplan Mark 
Published: November 2024

Abstract
Cytokine signaling is critical for cell-to-cell communication in development, homeostasis and disease. Interleukin (IL)-9 signaling promotes allergic diseases by acting on both immune and structural cells. In hematopoietic cells, IL-9 receptor (IL-9R) consists of two subunits – the α-chain (IL-9Rα) and a common γ (γc) chain. However, IL-9-responsive epithelial cells express IL-9Rα but not the γc chain raising a question on how these cells respond to IL-9 in the absence of the established IL-9R complex. Here, we discovered that in airway epithelial cells, IL-9Rα pairs with IL-13Rα1, forming a novel type II IL-9R. Using proximity ligation assay, we show that the IL-9Rα /IL-13Rα1 chains are in IL-9-dependent proximity in both mouse tracheal epithelial cells (mTEC) and human small airway epithelial cells (SAEC). IL-9-induced STAT3 activation in SAEC was blocked by anti-IL-13Rα1 but not by anti-γc. Using RNA-seq, in vitro IL-9 treatment of SAEC induced the expression of genes including CXCL3, a key chemokine implicated in asthma. Importantly, intranasal administration of IL-9 increased Cxcl3 expression in mTECs and CXCL3 protein in the lung, that corresponds with an expansion in the number of CXCR2+ immune cells in the lung. Thus, we identify a novel type II IL-9R which is composed of IL-9Rα and IL-13Rα1 and show that IL-9 signaling via type II IL-9R induces CXCL3 expression in epithelial cells to contribute to airway inflammation.
Tumor expressed BCAM impedes anti-tumor T cell immunity and can be targeted therapeutically
Authors: Flies Dallas , Tian Linjie , O'Neill Rachel , Fitzgerald Daniel , Sharee Sasan , Shaik Jahangheer , Bosiacki Jason , Paucarmayta Ana , Prajapati Kushal , Langermann Solomon , Mrass Paulus 
Published: November 2024

Abstract
The significant albeit limited clinical benefit of therapeutic targeting of checkpoint inhibitors highlights the need for immunological understanding of cancer escape from anti-tumor immunity to develop novel and better therapeutics. We surmise that many protein-protein interactions that regulate T cell immunity have yet to be identified. Using human gain-of-function genetic screens, thousands of proteins were screened for their ability to inhibit primary human T cell function. B Cell Adhesion Molecule (BCAM) was identified as a potent inhibitor of T cell proliferation and cytotoxic T cell (CTL) ability to kill human tumors in vitro. Tumor cell surface BCAM inhibited anti-tumor immunity promoted tumor growth in vivo. Blockade with decoy soluble BCAM protein fused to a either a silenced or functional Fc domain (BCAM Fc fusion protein) was able to promote T cell anti-tumor immunity and reduce tumor growth in syngeneic and humanized CDX tumor models. Moreover, tumor BCAM interaction with known ligand Laminin-a5 (LAMA5) resulted in increased production of LAMA5 that was associated with ECM modifications and potent exclusion of T cells from the tumor microenvironment (TME). BCAM Fc disruption of BCAM-LAMA5 interactions resulted in decreased LAMA5 production, increased T cell infiltration and decreased tumor growth. These data support a multi-pronged role for BCAM in anti-tumor T cell suppression and exclusion that can be targeted therapeutically.
A protective role of CD4+ T cells during Staphylococcus aureus craniotomy infection
Authors: Kak Gunjan , Van Roy Zachary , Horn Christopher , Heim Cortney , Kielian Tammy 
Published: November 2024

Abstract
Craniotomy is a neurosurgery performed to resolve severe neurological complications. Despite prophylaxis, nearly 1-3% of craniotomies are complicated by infection, with approximately half attributed to Staphylococcus aureus (S. aureus). Using our novel mouse model, we established a protective role for CD4+ T cells during S. aureus craniotomy infection. Rag1 knock out (KO) mice or animals treated with anti-CD4 Ab showed significantly elevated bacterial burden, suggesting an important role for adaptive immunity in controlling infection. These findings were corroborated in mice treated with VLA-4 and LFA-1 Abs, underscoring the importance of peripheral CD4+ T cell responses. scRNA-seq revealed phenotypic heterogeneity within brain CD3+ infiltrates with CD4+ cells being most predominant that were typified by robust inflammatory and glycolytic profiles. CD4+ T cells displayed attributes of both Th1 and Th17 phenotypes, whose functional importance was validated in vivo as adoptive transfer of either Th1 or Th17 cells prevented S. aureus outgrowth in Rag1 KO mice. Interestingly, milder phenotypes were observed in IL-17A/F or IFN-γ KO mice, suggesting a redundant and cooperative role of both Th1 and Th17 driven responses. This was confirmed by treatment of IFN-γ KO mice with IL-17A/F neutralizing Ab, which recapitulated phenotypes of Rag1 KO animals. Collectively, our results implicate a critical role for CD4+ T cells in S. aureus containment during craniotomy infection.
106
Authors: Jamsandekar Minal 
Published: November 2024

Abstract
Major histocompatibility complex (MHC) genes are among the most polymorphic and fastest evolving genes in vertebrate genomes with more than 1,000 alleles found at some human MHC loci. In this study, MHCII genes are explored in Atlantic herring, a species that comprises one of the biggest populations among vertebrates and occurs in huge shoals providing a potential target for pathogens using whole genome, long read sequencing of 14 individuals from three different geographic regions. The analysis identified a total of nine MHC class II loci present on three chromosomes. Some of the genes show extremely high nucleotide diversity (π = 10-13%) compared with the genome average (0.3%) as well as strong signatures of positive selection (dN/dS &gt;&gt; 1). Furthermore, we showed a highly significant nonrandom association of alpha and beta genes, strongly suggesting coevolution of alpha and beta alleles forming the peptide binding domain of MHC class II molecules. To the best of our knowledge this is the first genetic study of MHC polymorphism in a non-model organism based on whole genome, long-read sequencing. It demonstrates that the Atlantic herring MHC class II genes are among the most polymorphic so far described. The very high degree of polymorphisms is most likely explained by the combination of the large population size minimizing loss of genetic diversity due to drift and pathogen-driven selection.
Deciphering the Role of LIGHT/TNFSF14 in Driving Inflammatory Activity in Human Lung Fibroblasts
Authors: Ghosh Nandita , Gupta Rinkesh , Fung Kai , Croft Michael 
Published: November 2024

Abstract
LIGHT/ tumor necrosis factor superfamily member 14 (TNFSF14), is a protein that we previously found was central to lung inflammation and tissue fibrosis in mouse models of severe asthma and systemic sclerosis. We have hypothesized that one of its cellular targets involved in airway remodelling is the lung fibroblast given that both of its receptors, LTβR and HVEM are strongly expressed by these cells. With RNA-seq, we investigated the activity of LIGHT in these cells compared to other important inflammatory cytokines involved in remodeling in lung diseases, namely IL13, IL17A, and TGFβ. LIGHT induced a distinct set of inflammatory gene transcripts in human lung fibroblasts, including the adhesion molecules ICAM-1 and VCAM-1; cytokines IL32, IL33 and IL34; chemokines CXCL1, CXCL5, CXCL6 and CXCL10; and matrix metalloproteinases MMP9 and ADAM8. LIGHT with IL13 synergistically induced CCL2, CCL26, IL32 and ADAM8. LIGHT with IL17A also strongly boosted expression of several chemokines CXCL1, CXCL2, CXCL5, and CXCL6. Lastly, in fibroblasts differentiated into myofibroblasts by TGFβ pre-treatment, LIGHT enhanced the expression of many inflammatory and profibrotic transcripts such as ITGA1, ITGA11, MYH11, and Col1A1. This study highlights the unique as well as synergistic activities of LIGHT on these cells and suggests it could drive strongly inflammatory and remodeling effects in these cells relevant for several diseases dependent on the cytokine environment found in the lungs.
Immunomodulation of Dendrobium fimbriatum against blue light-induced ocular inflammation
Authors: Chang Wei-Chin , Liu Chuan-Teng , Song Ying-Chyi , Heinrich Michael , Yen Hung-Rong 
Published: November 2024

Abstract
As electronic device use increases, concerns about the harmful effects of blue light on eye health have grown. Past studies support the idea that Dendrobium species offer benefits such as reducing inflammation, providing antioxidants, and protecting against cataracts. We aim to investigate the immunomodulatory effect of Dendrobium fimbriatum Hook. extracts (DFs) in blue light-induced ocular damage. DFs decoctions were prepared from the fresh stems of 1-year, 2-year, and 3-year-old DFs. In vitro, cell viability, apoptosis, pro-inflammation cytokines and reactive oxygen species (ROS) were assessed. In vivo, ophthalmological and histological examination, systemic and local immunological changes were evaluated. Results showed that 2-year-old DF (DF-2yD) presents the maximal alleviation of blue light-induced cell death, apoptosis, interleukin (IL)-6 and 8, and ROS in retinal and corneal cells. In blue light-induced ocular damage mouse model, DF-2yD potently attenuated the fundus abnormality and retinal thickness. Furthermore, DF-2yD decreased IL-6 and TNF-alpha production in the draining lymph nodes. Collectively, the results indicate that DF hold promise as potential agents to counteract blue light-induced eye damage through the immunoregulation.
T cell responses in pediatric patients with severe influenza disease.
Authors: S. Menon Smrithi , Souquette Aisha , Allen E Kaitlynn , St. James Deryn , Mettelman Robert , Clark Brandi , Chase Crawford Jeremy , Novak Tanya , Randolph Adrienne , Thomas Paul 
Published: November 2024

Abstract
Influenza virus infections impose a high risk of disease-related complications in children leading to hospitalization, and increased mortality. However, it is still unclear why only some children become critically ill. The Pediatric Intensive Care Influenza 2 study enrolled children and young adults with influenza-related critical illness, aiming to identify predictive and prognostic biomarkers of severe disease (SD). To this end, we characterized T cell populations and functional responses to influenza, to understand how adaptive immunity fails to protect patients in this cohort from SD. Our preliminary studies indicate an increased level of ICOS expressing circulating T follicular helper cells (cTfh) in patients with SD while an elevation in the levels of effector memory CD4 and CD8 T cell populations in patients with moderate disease. Moreover, an enhanced level of IL21, CXCR3, ICOS and PD1 producing cTfH cells was observed in SD patients within the age group of 9-19 yrs. Age dependent varying expression of IL2, granzyme B producing CD4 T cells, CXCR3, ICOS producing and IL21 expressing cTfH cells was also observed. Additional T cell characterization is underway to integrate the influence of patient parameters such as vaccination status. T cell repertoire profiling and assessment of epitope-specific T cell populations using single-cell sequencing is being conducted to understand how pre-existing anti-influenza immunity affects disease severity and outcome.
IL-17A-mediated JMJD3 regulation of Cxcl1 gene expression in diabetic keratinocytes increases neutrophil recruitment and impairs wound repair.
Authors: Moon Jadie , Wolf Sonya , Audu Chris , Melvin William , Joshi Amrita , Mangum Kevin , Barrett Emily , Bauer Tyler , Shadiow James , Mazumder Purba , Rocco Sabrina , Erturk Ilkim , Saldana de Jimenez Gabriela , Moore Bethany , Davis Frank , Obi Andrea , Gallagher Katherine 
Published: November 2024

Abstract
Diabetic keratinocytes display an overexpression of chemokines that contribute to prolonged inflammation and non-healing. Using sequencing, we identified that Th17 cells and an IL-17A signature predominate in human diabetic wound tissue compared to non-diabetic controls. The effects of increased IL-17A in diabetic skin on keratinocyte function is unknown. We subjected isolated murine keratinocytes to a 5hr rIL-17A stimulation and performed bulk RNA-seq. We observed increased neutrophil chemoattract genes Cxcl1, Cxcl3, Cxcl5 compared to their controls. We analyzed epigenetic enzymes known to increase gene expression and identified that JMJD3, an activating demethylase, was increased both in human and murine diabetic keratinocytes. IL-17A stimulation increased Jmjd3 expression in murine keratinocytes, and keratinocytes from mice with a keratinocyte-specific IL-17RA deficiency were unable to increase Jmjd3 with rIL-17A. Treatment with a JMJD3-specific inhibitor decreased chemokine genes in IL-17A-stimulated keratinocytes. Keratinocytes from mice with a keratinocyte-specific JMJD3-deficiency displayed decreased chemokines compared to their littermate controls with rIL-17A, suggesting that JMJD3 is relevant to IL-17A-mediated regulation of chemokines. We then analyzed immune cells recruited to the wounds of normal and diabetic mice via flow cytometry and found that Ly6G+ neutrophils were significantly increased in diabetic mice on day 5, a timepoint at which IL-17A is elevated.
Cellular landscape of the esophageal epithelium in systemic sclerosis
Authors: Dapas Matthew , Clevenger Margarette , Makinde Hadijat , Therron Tyler , Wei Cenfu , Carns Mary , Aren Kathleen , Carlson Dustin , Muhammad Lutfiyya , Pandolfino John , Perlman Harris , Winter Deborah , Tetreault Marie-Pier 
Published: November 2024

Abstract
Systemic sclerosis (SSc) is a rare autoimmune disease characterized by vasculopathy and progressive fibrosis of the skin and internal organs. Individuals with SSc often suffer from chronic acid reflux and dysphagia due to loss of esophageal motility, but this pathogenesis is poorly understood.
Recently, studies have suggested that esophageal epithelial cells (EECs) may play a central role in the pathogenesis of SSc esophageal dysmotility. In this study, we performed a thorough transcriptomic investigation of the SSc esophageal epithelium in humans to determine whether distinct changes in EECs contribute to esophageal impairment in SSc.
We performed single-cell RNA sequencing of paired proximal and distal esophageal mucosa biopsies from 10 individuals with SSc, 4 comparator individuals with gastroesophageal reflux disease (GERD), and 6 healthy controls (HCs), yielding 230,720 EECs across 40 samples. SSc and GERD samples had significantly fewer terminally differentiated, superficial cells than HCs, and differential gene expression between conditions was primarily limited to superficial EECs. Gene dysregulation in SSc was highly correlated with GERD, but was relatively greater in the proximal region, including unique dysregulation of immune mediators that correlated with esophageal dysmotility.
This work sheds light on the cellular roots of esophageal dysfunction in SSc and serves as an atlas to guide future efforts to identify actionable targets in the esophagus in SSc.
Cross-tissue single-cell landscape of neutrophil heterogeneity and immunity in health and disease
Authors: Zheng Hong , Maestri Evan , Khatri Purvesh 
Published: November 2024

Abstract
Neutrophils, the most abundant leukocytes in human blood, play multifaceted roles in health and disease. They serve as essential components of immune defense against infection while also contribute to pathogenesis in a variety of diseases including cancer and autoimmunity. Many studies have identified functionally diverse neutrophil subsets in specific diseases and used varied nomenclatures. A comprehensive landscape of neutrophil heterogeneity and immunity is not yet fully elucidated.
To fully characterize neutrophils in health and disease, we collected and curated the transcriptomic profiles of 910,791 neutrophils from over 14 tissues across 24 single-cell RNA-seq datasets to generate a neutrophil single-cell compendium, which covered multiple diseases including viral and bacterial infection, cancer, diabetes, cardiovascular disease, and autoimmune diseases. We identified multiple neutrophil subsets exhibiting distinct heterogeneity along the development trajectories from pro- and immature neutrophils to mature neutrophils. We focused on pro- and immature neutrophils as well as mature neutrophil subsets displaying immunosuppressive transcriptional programs shared across diseases such as cancer and infections. We also inferred the cell-cell interaction between neutrophils and other immune cells in various disease contexts. Together, our systematic meta-analysis of neutrophils supported the development of neutrophil-based diagnostic tools and immunotherapy strategies.