bluuebunny/crossref_metadata_2025_split huggingface dataset:
Homeostatic Rho GTPase activation governs thymopoiesis
Authors: Kinsella Sinéad , Evandy Cindy A , Cooper Kirsten , Iovino Lorenzo , Jain Reema , deRoos Paul , Dudakov Jarrod A 
Published: January 2023

Abstract
The thymus is extremely sensitive to damage, resulting in attenuated T cell reconstitution and failure to elicit appropriate responses to infection, cancer and immunization. Although the thymus has a remarkable regenerative capacity, the mechanisms underlying this endogenous regeneration remain poorly understood. Previously we have identified two critical secreted factors that drive thymic regeneration, including BMP4 from endothelial cells (ECs) and IL-22 from dendritic cells (ILCs), which can be used as therapeutic regenerative strategies. Additionally, we have shown that steady-state CD4+CD8+ thymocytes (DPs) suppress levels of IL-23 (Science 336:91), a key regulator of IL-22; and as developing thymocytes undergo high levels of apoptosis, we hypothesize that apoptotic DPs mediate suppression of regenerative factors under homeostatic conditions, and damage-induced DP depletion removes this suppression, triggering thymic recovery. Here, we demonstrate a DP-specific reduction in cl-caspase 3 levels after acute injury in mice (total body irradiation, 550 cGy), and reduced Bmp4 expression in ECs co-cultured with apoptotic thymocytes, an effect rescued by apoptosis inhibition, using the pan-caspase inhibitor zVAD-FMK. We identified that apoptotic thymocytes mediate their suppression via TAM receptors in ECs and DCs, which subsequently activate Rho GTPases. Importantly, we show that inhibition of Rac1 activation increases Bmp4 and Il23 expression in ECs and DCs, respectively, and in vivo inhibition of Rac1 with EHT1864 enhances thymus cellularity in models of acute injury and age. These data describe a novel regulatory pathway that can be therapeutically targeted to boost T cell reconstitution after damage.
Ischemic Factors Precipitate a Shift in Immune Cell Populations at the Site of Cerebrovascular Blockage
Authors: Ortega Sterling B , Zevallos Cynthia B , Orozco Darko Q , Tranel Daniel , Karandikar Nitin J. , Ortega-Gutierrez Santiago 
Published: January 2023

Abstract
The incidence of stroke has risen over the past decade and will continue to be one of the leading causes of adult disability and death worldwide. Reperfusion therapy or endovascular thrombectomy (EVT) for large vessel occlusion (LVO) remains the only proven stroke therapy during the acute phase. But a select set of patients still exhibit poor recovery of neurological function, despite active cortical reorganization. The mechanisms that underlie this neurological dysfunction is still poorly understood. Subsequent to ischemic injury, a massive inflammatory response is seen for days, which exacerbates neuropathology in stroke models. Within the draining lymph nodes of ischemic patients, innate immune cells are actively phagocytizing brain antigen and presenting them to adaptive immune cells.
The focus of this project is to determine if early changes in immune cells near the site of occlusion can be used as predictors of long-term neurofunctional decline. Acute anterior circulation LVO patients treated with EVT will be enrolled in this study. At the time of thrombectomy, blood was collected proximal and distal to the thrombus prior to recanalization. As a control, blood was also collected from the femoral artery. Deep immune profiling of innate and adaptive immune cells using 13-parameter flow cytometry was performed revealing a specific increase in post-thrombus CD4 T-cells, as compared to pre-thrombus. Cytokine analysis revealed a decrease in pre- and post-thrombus IFN-γ and IL1α. Induction of in vitro hypoxia in healthy peripheral blood mononuclear cells revealed a similar reduction in IFN-γ.
We hypothesize that ischemia precipitates a shift in immune cells at the site of cerebrovascular blockage that modulates stroke recovery.
Chlorovirus Acceleration of Motor Neuron Disease in SOD1G93A Transgenic Mice may Clarify some of the Augmented anti-Chlorovirus Serology in Amyotrophic Lateral Sclerosis Patients
Authors: Petro Thomas M , Pattee Gary L. , Guinn Zachary P. , Agarkova Irina V. , Dunigan David D. , Van Etten James 
Published: January 2023

Abstract
Infections and inflammatory responses may contribute to motor neuron diseases (MND), such as Amyotrophic Lateral Sclerosis (ALS). A portion of familial ALS is caused by polymorphic Superoxide Dismutase (SOD)1 gene (SOD1G93A). As chloroviruses (CV) infect macrophages inducing inflammatory cytokines, they may contribute to ALS. Moreover, ATCV-1 is one of several chlorovirus types that encodes its own SOD1. Thus we evaluated serum from human ALS patients compared with controls for antibody against ATCV-1. In addition, we i.c. injected saline or ATCV-1 into SOD1G93A-transgenic and C57Bl/6 control mice and assessed development of ALS-like MND. Sera from ALS patients and controls had antibody to ATCV-1. However, ALS patients had significantly greater IgG1 anti-ATCV-1 compared with controls. This was accompanied by elevated IL-6, IFN-γ, and anti-Tetanus toxin antibody, but not IL-17. SOD1G93A mice inoculated with ATCV-1 had significantly accelerated development of MND, including tail paralysis, hindlimb tucking, decreased righting reflex, and latency to fall in a hanging cage lid test, compared with saline injected SOD1G93A mice. Splenic T cells of SOD1G93A and control mice showed similar IFN-γ and IL-17 responses to anti-CD3/-CD28, while Foxp3 expression overall was significantly higher in T cells of SOD1G93A compared with control mice. Inoculation with ATCV-1 decreased Foxp3 expression in SOD1G93A T cells compared with mice not given ATCV-1. These results indicate that chloroviruses, ubiquitous in fresh waters worldwide, induce antibody responses in humans that are elevated in ALS and may accelerate MND in the SOD1G93A type of familial ALS. Funding was generously provided by the Stuart Nichols ALS Research Foundation.
In utero exposure to endogenous maternal polyclonal anti-Caspr2 antibody leads to an autism-like phenotype in male mice
Authors: Bagnall-Moreau Ciara N , Huerta Patricio , Volpe Bruce , Diamond Betty , Brimberg Lior 
Published: January 2023

Abstract
Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder with a complex etiology. A strong sex bias has been observed in ASD with boys being four times more likely to be diagnosed than girls. The maternal immune system has been repeatedly implicated in the etiology of ASD. Maternal infection with fever during pregnancy has been linked to ASD. Increased levels of maternal cytokines and chemokines during gestation, including interleukin 6, have been associated with ASD in children with intellectual disabilities. Studies, including our own, have consistently supported the hypothesis that maternal brain-reactive antibodies can predispose to ASD by affecting the developing brain during a critical period. We have previously demonstrated that antibodies targeting the neuronal protein Caspr2 are present at high frequency in mothers with brain-reactive serology and a ASD child. Furthermore, male mice but not female mice exposed in utero to a monoclonal anti-Caspr2 antibody cloned from a mother of a child with ASD, develop an ASD -like phenotype. In the present study we use a new model in which female mice are immunized with the extracellular portion of Caspr2, and therefore harbor anti-Caspr2 antibodies throughout gestation to mimic the human condition. Male mice born to dams harboring polyclonal anti-Caspr2 antibodies exhibit abnormal cortical development, decreased dendritic complexity of excitatory neurons and reduced numbers of inhibitory neurons in the hippocampus, as well as repetitive behaviors and impairments in social interactions as adults. These studies will increase our understanding of the triggers of neurodevelopmental impairment and possibly suggest novel approaches to protecting the fetal brain.
Development of a platform for screening patient-derived CD8+ T cell responses against autoantigens
Authors: Johnson Renee K , Clarkson Ben D.S. , Howe Charles L 
Published: January 2023

Abstract
Cytotoxic CD8+ T cells outnumber CD4+ T cells within multiple sclerosis (MS) lesions, yet the pathogenic role for these cells and the relevant target antigen(s) involved in neuropathology remain unknown. We predict that a subset of lesion-infiltrating CD8+ T cells are specific for neuronal antigens and contribute to axon or neuron injury, thereby driving disease progression and accumulation of disability. To test this, we are developing protocols to measure MS patient-derived T cell reactivity against neuronal and control antigens. Peripheral blood mononuclear cells (PBMCs) isolated from whole blood were incubated with GM-CSF and IL-4 to selectively expand dendritic cells (DCs) or with IL-7 to promote survival and maintenance of T cells. After resting, patient T cells were combined 1:1 with autologous DCs in the presence of vaccine antigens or peptides derived from common pathogens. We tested responses against a pool of cytomegalovirus, Epstein Barr virus, and influenza peptides (CEF) and against mumps viral protein extract. We also tested responses in T cells derived from patients with autoimmune neurological disorders following incubation with autologous DCs and disease-specific proteins. Activation profiles were assessed using CD69, CD44, and LFA-1 by conventional flow cytometry. Cellular responses were also assessed using mass cytometry (cytometry by time of flight; CyTOF). Our next steps include testing MS patient-derived T cell responses against neuronal and axonal protein libraries, with the goal of identifying potential autoantigens that underlie disease progression.
Cytotoxic T cells infiltrate the brain and interact with microglia to reduce Alzheimer’s Disease pathogenesis
Authors: Sanchez Jessica Ramirez , Marsh Samual , McIntyre Laura , Davtyan Hayk , Walsh Craig , Blurton-Jones Mathew 
Published: January 2023

Abstract
The role of the adaptive immune system, specifically T cells, remains poorly understood in Alzheimer’s disease (AD) pathogenesis research. Our lab generated an immune-deficient mouse model of AD (RAG2ɣc−/−5xfAD) which showed a dramatic increase in amyloid plaque load, an activated microglial neuroinflammatory state, and a decrease in the ability of microglia to phagocytose beta-amyloid. Restoration of adaptive immunity via bone marrow transplantation led to a corresponding decrease in amyloid plaques, indicating a fundamental role for adaptive immunity in AD pathology. Further adoptive transfers into these aged immune-deficient mice showed a significant increase in memory CD8+ T cells and microglia in the RAG2ɣc−/−5xfAD brain parenchyma via flow cytometry. Interestingly, T cells were observed adjacent to plaque-associated microglia in these brains via Immunohistochemistry. To determine whether T cell infiltration also occurs in immune-intact AD mice, flow cytometry was conducted on 5xfAD mice that develop amyloid plaques, PS19 mice that develop neurofibrillary tangles, PS19-5xfAD bigenic mouse model, and wild-type mice. This analysis validated the immune-deficient observations, demonstrating a significant increase in memory CD8+ T cells in 5xfAD mice brains but also revealed a further significant increase in the bigenic PS19-5xfAD model. Therefore, I hypothesize that cytotoxic T cells respond to the inflammatory AD environment by infiltrating the brain and directly interacting with microglia to decrease amyloid plaques. My initial data clearly demonstrate that further examination of the crosstalk between adaptive and innate immunity is needed to determine the mechanisms by which T cells influence AD pathogenesis.
Single-Cell RNA Sequencing Identifies a Novel Population of CD4+CD8+ T Cells that are Regulated by COX-2 During Allergic Lung Inflammation
Authors: Li Hong , Bradbury J Alyce , Xu XiaoJan , Edin Matthew L , Li Jianliang , Katen Kevin , Gruzdev Artiom , Graves Joan P , Duval Caroline N , Zeldin Darryl C 
Published: January 2023

Abstract
To better understand the relative contributions of different cell types during allergic lung inflammation and the role of cyclooxygenase (COX)-derived eicosanoids in this process, we compared vehicle (control) and ovalbumin (OVA)-exposed lungs from wild type (WT) and COX-2−/− mice by single-cell RNA sequencing (scRNA-Seq). Twenty-two unique cell clusters were identified in allergic lungs based on cell signature markers. In addition to known cell types, we identified 2 clusters of CD4+CD8+. Both populations were mitotically active (high % in S/G2/M phases) and exhibited a unique molecular identifier (UMI) index. FACS analysis confirmed the presence of these two distinct cell populations (CD4+HCD8+H and CD4+LCD8+H) that were increased after OVA-induced allergic lung inflammation. Interestingly, isolated CD4+HCD8+H T cells stimulated with phorbol ester expressed IL-4, IL-5 and IL-13, but not IFNγ. In contrast, isolated and stimulated CD4+LCD8+H T cells primarily expressed IL-5. Mass cytometry (CyTOF) using a 37-antibody panel confirmed the increase in CD4+CD8+ T cells after OVA sensitization/exposure, as well as the induction of IL-4, IL-5 and IL-13. Importantly, CD4+CD8+-1 and CD4+CD8+-2 cell populations identified by scRNA-Seq were increased in lungs from OVA-treated COX-2−/− mice compared to lungs from OVA-treated WT mice. Furthermore, COX-2−/− CD4+CD8+ T cells preferentially produced more IL-4 and IL-5 than WT CD4+CD8+ T cells. Thus, using scRNA-Seq, CyTOF and FACS analyses, we identified two populations of CD4+CD8+ T cells that produce immunoregulatory cytokines and are regulated by COX-2. These cells may play an important, previously unappreciated role in the pathogenesis of allergic lung disease.
Blockade of RANKL/RANK interaction prevents allergic airway inflammation in a mouse model of asthma
Authors: Maslanka Tomasz , Gregorczyk Izabela , Jasiecka-Mikolajczyk Agnieszka 
Published: January 2023

Abstract
W are constantly looking for new therapeutic options for asthma treatment. In this study, we investigated whether the signaling pathway of the receptor activator of nuclear factor (NF)-κB (RANK), RANK ligand (RANKL), and its decoy receptor osteoprotegerin (OPG) may be considered as a novel drug target for therapy of allergic asthma. Therefore, we tested the effect of blockade of RANK-L with OPG on the development of ovalbumin (OVA)-induced airway allergic inflammation (AAI) and accompanying immune response. The absolute counts of CD11chighCD11b+CD103− (CD11b+) dendritic cells (DC), ST2+Foxp3−CD4+ (Th2) cells and BrdU+(i.e., proliferating)CD4+ T cells were determined in the mediastinal lymph nodes (MLNs) and lungs of untreated non-immunized mice, untreated OVA-immunized mice and OPG-treated OVA-immunized mice. Immunization with OVA induced a significant increase in the mean scores of lung histologic lesions and administration of OPG completely prevented this effect. The absolute counts of CD11b+ DC, Th2 cells and BrdU+CD4+ T cells were increased in the MLNs and lungs of untreated OVA-immunized mice. Treatment with OPG attenuated, but did not abolish, the OVA-induced increase in the count of CD11b+ DC in the MLNs. In turn, administration of this agent prevented the OVA-induced increase in the count of Th2 cells and BrdU+CD4+ T cells in the MLNs and lungs. In conclusion, blockade of RANK/RANKL interaction prevents OVA-induced AAI and this effect is mediated through inhibition the allergen-induced clonal expansion of CD4+ T cells in the MLNs. Clinical implication: the local inhibition of RANK/RANK-L interaction within the respiratory system can be considered as a novel therapeutic strategy in treatment of allergic asthma.
IgM regulates airway hyperresponsiveness in allergic asthma through acetyl choline receptors
Authors: Hadebe Sabelo , Khumalo Jermaine , Ngomti Amkele , Mangali Sandisiwe , Scibiorek Martyna , Brombacher Frank 
Published: January 2023

Abstract
Allergic asthma is a disease driven by T helper 2 (TH2)-type cells secreting interleukin (IL-) 4, 5 and 13. It is characterized by eosinophils, airway hyperesponsiveness (AHR) and IgE secreting B cells. B cells play a role in allergic asthma in an allergen load dependent manner. IgM isotype secreted by naïve B cells is important for class switching. It is currently unclear how IgM isotype contributes in the development of allergic asthma. We investigated the role of IgM isotype in a house dust mite (HDM)-induced TH2 allergic asthma model. We sensitised wild type (wt), IgM-deficient (IgM−/−) and B cell-deficient (uMT−/−) mice with high (&gt;10 ug) and low (&lt;3 ug) dose of HDM extracts. We found IgM to be essential in antibody class switching and in AHR, but not in TH2 airway inflammation or eosinophilia. RNA seq suggested that IgM regulated AHR through modulating muscarinic receptor genes, particularly M3 and this was independent of microbiota. Transfer of wild type serum, but not B cells into IgM−/− mice could restore class switching but not AHR. These unprecedented findings suggest that IgM has a unique function in allergic asthma and regulates AHR by interacting with non-immune genes.
Resp-Aid in Ova-challenged mice decreased airway inflammatory cell infiltration, AHR and IL-13 cytokine expression levels
Authors: Nascimento Amanda AN , Sandini Thaisa Meira , Thompson Brooke , Schneberger David , Bernardi Maria Martha , Singh Baljit , Kirychuk Shelley 
Published: January 2023

Abstract

Introduction
Asthma is an inflammatory disease caused by repeated immediate phase hypersensitivity and late-phase allergic reactions in the lungs. Resp-Aid is a formula composed of 9 medicinal herbs that supports in the resolution of respiratory disorders (Coltsfoot, Marshamallow, Plantain, Mullein, Horehound, Licorice, Lobelia, Ginger, and Myrrh). These herbs, alone, have shown positive effects in reducing symptoms associated with asthma. However, the effects of the combination of these herbs on asthma is not well understood. Therefore, we conducted a series of studies to explore the mechanism of action of Resp-Aid.

Material and Methods
C57Bl/6 male mice (n = 7 each group) were challenged with or without ovalbumin (OVA). After the challenge, they were divided into 4 groups where 2 groups received 1 drop of Resp-Aid twice daily for 7 days and 2 groups acted as control. On day 5 of the treatment with Resp-Aid the airway hyperresponsiveness (AHR) was measured. After the treatment with Resp-Aid, lungs and bronchoalveolar lavage (BAL) fluid were collected and the cytokines (IL-4, IL-5, IL-13, IL-17a, IFNy), MPO and EPO were quantified.

Results
All OVA-challenged mice had increased AHR, leukocytes and cytokine levels compared to their respective controls showing the development of an asthmatic response. The Resp-Aid asthmatic groups had decreased AHR, IL-13, neutrophils and EPO levels compared to asthmatic mice.

Conclusion
This data show that Resp-Aid changed the immune response related to asthma, and it may be a potential target for therapeutic intervention in allergic lung disease or asthma.
Paneth cells regulate allergic sensitization in the gastrointestinal tract
Authors: Kim Eunsoo , Burnett Drew J. , Joldrichsen Marisa R , Mori-Akiyama Yuko , Cormet-Boyaka Estelle , Boyaka Prosper N 
Published: January 2023

Abstract
The gut immune homeostasis plays a major role in controlling sensitization to food allergens and the development of allergic responses in the gastrointestinal tract, but also to skin or lung. Epithelial cells sensing of commensal microbes is crucial for the maturation of the gut immune system and antibiotic treatment during early life is believed to have contributed to the increased incidence of food allergy. Paneth cells secrete antimicrobial products and cytokines in the crypts of the small intestine, but their role in food allergy remains elusive. We found that SOX9ΔIEC mice, which lack of Paneth cells, display dysbiosis with increased microbial diversity. Interestingly, Paneth cell deficiency have increased numbers of ILC2 in the intestinal lymphoid tissues. Upon oral sensitization with a model food antigen in the presence of cholera toxin, SOX9ΔIEC mice developed significantly higher allergen-specific serum IgE responses, and enhanced signs of allergic responses (i.e., swelling, hypothermia, impairment of lung functions) after nasal allergen challenge. Transplantation of fecal materials from SOX9ΔIEC mice was able to transfer the hyper-allergenic phenotype to control wild-type mice and bacteria-free fecal extract from SOX9ΔIEC mice increased the number of IgE producing B cells than fecal extract from control mice with treatment of anti-CD40 and IL-4 in vitro. Ongoing studies are assessing the metabolites present in the intestinal lumen of SOX9ΔIEC mice and their effect of intestinal epithelial cells and myeloid cells to elucidate the mechanisms by which impaired or defective anti-microbial functions of Paneth cells regulate allergic sensitization of the GI tract.
Tracking the dynamics of the peanut-specific CD4+ T cell response in humans reveals novel disease-associated subsets
Authors: Lozano-Ojalvo Daniel , Berin M. Cecilia 
Published: January 2023

Abstract
Peanut allergy is associated with the presence of CD154+ Th2A and Tfh13 cells. The use of a single activation marker such as CD154 at a single time-point after stimulation, may bias the detection of terminal effector cells. Our objective was to track the dynamics of T cell in response to peanut to obtain a more comprehensive picture of allergen-specific immunity associated with disease.
PBMCs from peanut allergic (n=65), food allergic but peanut tolerant (n=5), and healthy controls (n=3) were incubated with a peanut extract for 6–48 h. Cell activation was monitored by conventional and high dimensional flow cytometry (30-marker panel) and cytokine production by cytokine capture assay. Analysis was performed by 2-D gating and unsupervised clustering (FlowSOM).
Peanut induced an upregulation of CD154/CD69 on mature effector memory cells that was observed at 6h and maintained for 48h. Cytokine capture showed secretion of IL-2, -4, -13, and -5 from these cells, which peaked at 6 h. The use of additional timepoints and activation markers identified two additional subsets specifically associated with disease: CD137+ and OX40/CD25 on other two cell subsets after 24 h. CD137+ cells secreted IFN-g, while OX40/CD25 cells secreted none of the cytokines tested. OX40/CD25 cells were reduced by IL-2 neutralization and likely include bystander activated cells. Clustering by FlowSOM analysis confirmed the identity of these three unique clusters, and provided additional phenotypic markers for CD154+ (CCR4+/CCR6+), CD137+ (CXCR3+), and CD25/OX40 (Foxp3+CD27−).
Analysis of the dynamics of peanut-induced T cell activation revealed novel subsets including Th1 and regulatory cells co-existing with previously described highly differentiated Th2 cells.