bluuebunny/crossref_metadata_2025_split huggingface dataset:
Examining the outcomes of enhanced CD28 signaling in vivo using ‘humanized’ mice
Authors: Brady Alexander , McGeachy Mandy 
Published: November 2024

Abstract
CD28 signaling is stronger in humans than in mice, as demonstrated by the phase one clinical trial of a CD28 superagonist antibody. In mice CD28 superagonist expanded regulatory T cells and reduced autoimmune disease, but in humans it induced life-threatening cytokine storm. The final signaling domain in human CD28 is PYAPP, while in mice it is PYAPA. We used CRISPR/Cas9 to develop a ‘humanized’ mouse with a CD28 A-&gt;P substitution (CD28A210P) to examine the outcomes of this variation on T cell-driven disease. In vitro, CD28A210P T cells had increased signaling, cytokine production, and proliferation. In vivo, after injection with CD28 superagonist, CD28A210P mice had increased weight loss, cytokine production, and less Treg expansion, supporting that humanized CD28 signaling could contribute to the excessive inflammatory response. Following LCMV cl13 infection, CD28A210P mice display enhanced early Th1 and CD8 T cell responses with exacerbated weight loss but then underwent increased exhaustion, leading to rapid recovery of weight. Similarly, CD28A210P mice show increased EAE incidence but milder symptoms, coinciding with an increase in CNS-infiltrating CD8 T cells which play a regulatory role in EAE. This study shows enhanced CD28 signaling due to an amino acid variation can alter T cell driven disease and reveals nuances in immunoregulation. This expands our knowledge of CD28, with potential implications for immunotherapeutics that target or utilize CD28 signaling.
Antigen-specific nanoparticle tolerance reduced inflammation at the target tissue and increased target protein expression in recombinant adeno-associated virus (AAV)-mediated gene therapy
Authors: Podojil Joseph , Xu Dan , Chiang Ming-Yi , Kakade Sandeep , McCarthy Derrick , Elhofy Adam , Miller Stephen 
Published: November 2024

Abstract
Recombinant Adeno-associated virus (AAV)-mediated gene therapy is an attractive approach for treating diseases where expression of an endogenous protein is defective or absent. While AAV is considered to be non-integrating and weakly immunogenic, AAV delivery elicits antigen (Ag)-specific T cells targeting transduced cells, and neutralizing antibodies inhibiting secondary administration. Effector T cell responses and vector dilution lead to decreased protein expression within target tissue, thereby requiring periodic AAV re-dosing with AAV to sustain the therapeutic benefit. Treatment with Ag-containing biodegradable poly(lactide-co-glycolide) (PLGA) nanoparticles (denoted as CNPs) represents a putative co-treatment allowing for decreased therapeutic AAV dose and frequency of AAV re-dosing to maintain therapeutic benefit. In the present studies, AAV8-eGFP was administered intramuscularly or intravenously in the presence or absence of B cell depletion. Prophylactic or therapeutic treatment with CNP(VP1 + eGFP) allowed for higher levels of eGFP expression within the target tissues following primary and secondary AAV8-eGFP dosing. Additionally, Ag-specific CNP(VP1 + eGFP) treatment inhibited the increase in effector CD8+ T cells within the target tissue following secondary AAV8-eGFP administration. Taken together, co-treatment of gene therapy patients with Ag-specific CNPs may allow for enhanced transgene expression over time, while allowing for AAV re-dosing.
Exploring cGAS-STING Pathway in Sjögren’s Disease: Driver of IFN production and Potential Therapeutic Target
Authors: Yamada Eiko , Abed Mehdi , Perez Paola , Jang Shyh-Ing , Warner Blake 
Published: November 2024

Abstract
Sjögren's Disease (SjD) is a prevalent autoimmune disorder characterized by dysfunctional lacrimal and salivary glands (SG) and lacks an effective therapy. Despite unknown pathogenic mechanisms, elevated interferon (IFN) expression is observed in SjD patients. Recent findings indicate activation of the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway in systemic lupus, prompting investigation into its role in SjD. We hypothesize cGAS-STING pathway activation in SjD contributing IFN dysregulation and symptoms. Bulk and single-cell RNA sequencing of SjD SG shows elevated IFN-related genes in epithelial and inflammatory cells. Investigation of the affected human SG by flow cytometry confirms upregulated phosphorylation status of cGAS-STING proteins in epithelial and immune cells; and epithelial cGAS-STING pathway activation directly correlates with the level of immune cell infiltration. Treatment of patient’s SG and peripheral blood mononuclear cells (PBMC) with a STING antagonist ex vivo reduces phosphorylated protein levels without cytotoxicity. These findings support our hypothesis that targeting the cGAS-STING pathway holds promise for mitigating salivary gland inflammation in SjD, preventing chronic inflammation's secondary effects and resulting tissue destruction.
Peripheral Natural Killer Cell Cytotoxicity is Suppressed in Amyotrophic Lateral Sclerosis
Authors: Murdock Benjamin , Teener Samuel , Webber-Davis Ian , Monteagudo-Caballero Alina , Famie Joshua , Piecuch Caroline , Goutman Stephen , Feldman Eva 
Published: November 2024

Abstract
Amyotrophic lateral sclerosis (ALS) is a deadly neurodegenerative disease where the immune system contributes to progression, though the underlying immune mechanisms are unclear. Natural killer (NK) cells likely play a critical role in disease progression: they are responsible for dispatching damaged cells, and during ALS they accumulate in the CNS and increase the rate of neuronal death. Thus, we hypothesize that NK cells are more cytotoxic in ALS patients than in healthy controls. To test this, primary NK cells were isolated from the blood of control and ALS patients and co-cultured for four hours at a 1:1 ratio with K-562 cancer cells. K-562 cell death was measured via e500 viability dye staining and used to quantify NK cell cytotoxicity. Surprisingly, we found that NK cell cytotoxicity was significantly reduced in ALS NK cells compared to healthy controls. Next, using flow cytometry, we measured a battery of peripheral inflammatory markers and correlated these measures with NK cell cytotoxicity to identify immune pathways impacting NK cell cytotoxicity. Unsurprisingly, in healthy control patients there was significant correlation between NK cell markers and NK cell cytotoxicity. However, these correlations were absent in ALS patients. Together, these data suggest that NK cell cytotoxicity is downregulated in ALS, likely as a mechanism to slow NK cell-mediated damage to motor neurons, though further studies will be required to elucidate the underlying mechanisms.
Isolation and characterization of epidermal Langerhans cells from human skin
Authors: Yamada Minami , Hasegawa Tatsuya 
Published: November 2024

Abstract
The skin is the largest organ that acts as the protective interface between the body and the environment. The immune system of the skin provides the first line of defense against continuous exposure of various external insults and pathogens. In the outermost layer of the skin, epidermal Langerhans cells (LCs) play a sentinel role as tissue-resident antigen presenting cells to confront the environmental insults. Ontogeny and functional studies of murine LCs have provided fundamental insights into LC biology, however, the nature of LCs and the immunological microenvironment surrounding LCs are different between mice and human skin. Therefore, it is important to study the LC biology in human skin. Here, we optimized a protocol for the isolation of epidermal LCs from sun-protected normal human skin tissues. Flow cytometry and immunocytochemistry staining analysis demonstrated that CD1a and CD207, which are human LC markers, were expressed in isolated epidermal LCs. In addition, we observed that epidermal LCs isolated from human skin highly expressed CD1a and CD207 in comparison with LCs derived from CD34+ hemopoietic stem cells. Our optimized protocol for isolating human epidermal LCs may help to fully capture the nature and role of human LCs in healthy and diseased condition.
CSF1R blockade in fructose-fed rhesus macaques improves clinical parameters of prediabetes.
Authors: Stahr Natalie , Phou Amy , Glover Colin , Perkins Jeanne , Haupt Erin , Palmer Clovis , Ardeshir Amir , Kim Woong-Ki 
Published: November 2024

Abstract
We have previously found that depletion of CSF1Rhi macrophages (Mθ) using a rhesus IgG1 recombinant antibody against CSF1R (α-CSF1R) reduced total cholesterol and improved insulin sensitivity in naturally prediabetic rhesus macaques (RM). High fructose diet (HFD)-fed RMs have been established as a model of metabolic syndrome, with increased insulin resistance (IR) and increased monocyte chemoattractant protein 1 (MCP-1). We hypothesize that depletion of CSF1Rhi tissue Mθ using a monoclonal α-CSF1R and replacement with freshly differentiated Mθ will alleviate inflammation in tissues and ameliorate prediabetic disease in HFD-fed RMs.
We induced IR and heightened hbA1C, indicating prediabetes in eight aged Indian RMs through administration of a HFD for 6 months. RMs were treated with either α-CSF1R or an IgG1 control administered intravenously (15 mg/kg) once every 2 weeks for 6 weeks. Clinical parameters such as plasma levels of total cholesterol, glucose, insulin, adiponectin, CSF1, IL1β, MCP-1 were monitored throughout treatment and up to 8 weeks after the 3rd injection. Mθ subsets were quantified by immunofluorescent (IF) staining.
Treatment with α-CSF1R resulted in decreased total cholesterol and insulin, increased adiponectin and CSF1. IF staining confirmed the depletion of CSF1Rhi macrophages in insulin sensitive tissues. Treatment with α-CSF1R resulted in improvement in multiple clinical parameters associated with prediabetes.
DUSP8 Promotes IL-9 Transcription by Inactivating Pur-α leading to asthma and atopic dermatitis
Authors: Chuang Huai-Chia , Tan Tse-Hua 
Published: November 2024

Abstract
Dual-specificity phosphatase 8 (DUSP8) is a member of the DUSP family. DUSP8 dephosphorylates and inactivates the kinase JNK. DUSP8 protein levels are predominantly expressed in CD4+ T cells and platelets. DUSP8 dephosphorylates and inactivates specifically JNK in Jurkat T cells; however, the in vivo role of DUSP8 in T cells remains unclear.
Using T-cell-specific DUSP8 conditional knockout (T-DUSP8 cKO, DUSP8f/f;CD4-Cre) mice, mass spectrometry, and chromatin-immunoprecipitation sequencing, we found that DUSP8 stimulated IL-9 gene expression and Th9 differentiation. TGF-β stimulated DUSP8 phosphatase activity in T cells. Mechanistically, DUSP8 dephosphorylated the transcriptional repressor Pur-α upon TGF-β signaling, leading to the nuclear export of Pur-α and subsequent IL-9 transcriptional activation. Furthermore, Il-9 mRNA levels in murine T cells were induced in Pur-α knockout. Reduction of IL-9 levels in T cells of T-DUSP8 cKO mice was reversed by Pur-α heterozygous knockout. Consistently, T-DUSP8 cKO mice displayed reduction of IL-9 and Th9-mediated immune responses in allergy, autoimmune responses, and anti-tumor immunity. Remarkably, DUSP8 overexpression and DUSP8-Pur-α interaction indeed occurred in the peripheral blood T cells of patients with asthma or atopic dermatitis, contributing to IL-9 overproduction in patients’ T cells. Thus, DUSP8 plays a critical role in the pathogenesis of asthma and atopic dermatitis, as well as autoimmune disease and cancer.
Requirement for Toll-like Receptor 4 (TLR4) in LPS-induced lung inflammation caused by atypical Cyanobacteria Lipopolysaccharide
Authors: Swanson-Mungerson Michelle , Monroy Del Toro Rosalinda , Incrocci Ryan , Williams Philip , Mayer Alejandro , Swartzendruber Julie 
Published: November 2024

Abstract
Cyanobacterial blooms found in freshwater sources cause multiple human diseases, including lung inflammation. Cyanobacteria produce atypical lipopolysaccharides (LPS) that can act as either weak TLR4 agonists or even antagonists in in vitro and in vivo models. However, our laboratory demonstrated that LPS from the cyanobacteria Geitlerinema sp. activates immune cells, despite containing altered LPS sugar side chains and inducing modified TLR4 signaling. Additionally, Geitlerinema sp. LPS induced lung inflammation in vivo based on significant neutrophil infiltration 24 hours post LPS exposure. In the current study, we extend our initial findings and asked whether the atypical LPS from an additional cyanobacterial strain, Brasilonema sp., also induces lung inflammation and if TLR4 is required. Our data demonstrate that in vitro, Brasilonema sp. LPS stimulates both Interleukin (IL)-6 and IL-8 production by immortalized airway epithelial cells that is TLR4-dependent, since the addition of the SPA-4 peptide blocks LPS-mediated production of these cytokines. More importantly, Brasilonema sp. LPS induced lung inflammation in wildtype mice but did not induce lung inflammation in TLR4-deficient mice, suggesting that the atypical LPS from cyanobacteria binds to TLR4 to induce lung inflammation. These findings help us to understand the impact of cyanobacteria LPS in neutrophil-dependent airway inflammation in individuals living near freshwater sources contaminated with cyanobacteria.
Food allergies as a promoter of transient anxiety-like behavior
Authors: Bustamante Hernandez Leslie 
Published: November 2024

Abstract
Food allergies impact human health, including mood. Allergen challenges in mice promote central amygdala activation, the integrative hub for anxiety and other mood disorders. Indeed, food allergic individuals show higher prevalence of anxiety. We hypothesize that food allergies induce anxiety. Female BALB/c mice received OVA/alum sensitization (or PBS/alum for control) subcutaneously on days 0 and 7, followed by five OVA oral challenges every three days from days 14 to 28. Behaviors were assessed using standard anxiety tests (open field test, OFT, elevated plus maze, EPM). Development of allergy was confirmed by increased specific IgE, anaphylaxis, and diarrhea. Allergic mice showed avoidance to open and elevated areas (EPM) when compared with the control group (p= 0.0425). We found that allergic animals showed less time spent in the center of the arena (p= 0.0008, OFT) compared with controls, suggesting the development of anxiety-like behavior. Because the locomotor activity (total distance and speed) was not different between allergic and controls, we confirmed a true anxiety-like phenotype induced by food allergy. Notably, we found that only one allergen challenge was enough to induce anxiety. However, this behavior was transient as there was no difference two weeks after the last allergen exposure. Our data suggest that this anxiety is dependent on allergen challenge and we propose that allergic inflammation might contribute to mental disorders.
The role of Per2 in atopic dermatitis experimental model
Authors: Kang Tae Jin 
Published: November 2024

Abstract
Atopic dermatitis (AD) is a chronic inflammatory skin disorder associated with defective skin barrier. The Clock gene Per2, one of circadian genes, has vital role in inflammation and it is up regulates inflammatory cytokines. In this study we studied the Per2 role in RAW 264.7 by transfecting with Per2 siRNA for knockdown specific Per2 gene and atopic dermatitis animal model using establishing DNCB induced Per2-/- mice. RAW 264.7 cells were simulated with LPS for 12 hours, then measured circadian and cytokine gene expressions. We found that PER2 induce the inflammation and enhance the expression of pro-inflammatory cytokines such as TNF-a and IL-6 by suppressing Bmal-1, which has anti-inflammatory activity, and PER2 is inducing nitric oxide (NO) level too. RAW 264.7 cells transfected with Per2 siRNA was not stimulated by LPS treatment and produced lower levels of inflammatory cytokines and NO by Bmal-1 up-regulation. We also examined activity of Per2 in DNCB induced atopic dermatitis animal model by using Per2+/+ and Per2-/- mice. We found Per2+/+ mice has higher number of Th2 cells, up-regulated Th2 cytokines and GATA3 expression, suppressed BMAL1 expression. Per2 -/- mice showed low levels of Th2 cytokines and IgE levels, less Th2 cells. We hypothesized PER2 has prominent role to induce inflammation and atopic dermatitis
162
Authors: Srivastava Neetu , Hu Hao , Peterson Orion , Vomund Anthony , Stremska Marta , Zaman Mohammad , Giri Shilpi , Li Tiandao , Lichti Cheryl , Zakharov Pavel , Zhang Bo , Abumrad Nada , Chen Yi-Guang , Ravichandran Kodi , Unanue Emil , Wan Xiaoxiao 
Published: November 2024

Abstract
Autoimmune disorders are characterized by destruction of specific tissues through invasive, antigen-specific responses. Yet, the influence of intrinsic tissue properties on susceptibility to autoimmunity remains uncertain. This study explored whether the proinflammatory features of resident macrophages in pancreatic islets could create an environment conducive to diabetic autoimmunity. Our analysis revealed elevated expression of CXCL16, a chemokine and scavenger receptor for oxidized low-density lipoproteins (OxLDL), in islet macrophages from both mice and humans, irrespective of autoimmune predisposition. Deleting Cxcl16 in NOD mice significantly suppressed the development of T1D. Mechanistically, the loss of Cxcl16 significantly impaired the capacity of islet macrophages to take up OxLDL. This coincided with OxLDL accumulation in pancreatic islets, leading to a reduction in intra-islet transitory (Texint) CD8 T cells displaying proliferation and effector signatures. Ferroptosis contributed to the reduction of these intra-islet Texint CD8 T cells, which are susceptible to oxidative stress. PD-1 blockade, however, rescued this cell population in the islets and reversed diabetes resistance in Cxcl16-deficient mice. Therefore, OxLDL scavenging by islet macrophages contributes to the local differentiation of pathogenic CD8 T cells. These findings suggest that the normal tissue microenvironment can facilitate autoimmune pathogenesis in predisposed individuals.
Oxidative stress is a shared characteristic of ME/CFS and Long COVID
Authors: Shankar Vishnu , Wilhelmy Julie , Curtis Ellis , Michael Basil , Cervantes Layla , Mallajosyula Vamsee , Davis Ronald , Snyder Michael , Younis Shady , Robinson William , Shankar Sadasivan , Mischel Paul , Bonilla Hector , Davis Mark 
Published: November 2024

Abstract
More than 65 million individuals worldwide are estimated to have Long COVID (LC), a complex multisystemic condition, wherein patients of all ages report fatigue, post-exertional malaise, and other symptoms resembling myalgic encephalomyelitis / chronic fatigue syndrome (ME/CFS). With no current treatments or reliable diagnostic markers, there is an urgent need to define the molecular underpinnings of these conditions. By studying bioenergetic characteristics of peripheral blood lymphocytes in over 16 healthy controls, 15 ME/CFS, and 15 LC donors, we find both ME/CFS and LC donors exhibit signs of elevated oxidative stress, relative to healthy controls, especially in the memory subset. Using a combination of flow cytometry, bulk RNA-seq analysis, mass spectrometry, and systems chemistry analysis, we observed gender-specific aberrations in ROS clearance and damage pathways. While ME/CFS females exhibit higher total ROS and mitochondrial calcium levels, ME/CFS males have normal ROS levels, but larger changes in lipid oxidative damage. The higher ROS levels correlate with hyperproliferation of T cells and can be attenuated by metformin, suggesting this FDA-approved drug may be an effective treatment. Thus, we report that both ME/CFS and LC are mechanistically related and can be diagnosed with quantitative blood cell measurements. We also suggest that effective, patient tailored drugs might be discovered using standard lymphocyte stimulation assays.
Pharmacological activation of LANCL2 provides immunometabolic support for the restoration of cognitive function markers in a mouse model of Alzheimer’s disease
Authors: Leber Andrew , Hontecillas Raquel , Tubau-Juni Nuria , Fitch Sarah , Bassaganya-Riera Josep 
Published: November 2024

Abstract
LANCL2 is a member of a three-protein family of receptors that acts as the mammalian receptor for abscisic acid (ABA). LANCL2 expression is significantly diminished both with the brains of 3xTg mice as well as human patients with Alzheimer’s disease. Immunologically, LANCL2 expression is closely associated with regulatory CD4+ T cells and their suppressive capacity as well as within microglia. Treatment of mice with ABA improved spatial memory and reduced amyloid beta load. To study the effects of LANCL2 activation in vivo, we created a novel agonist. After 12 weeks of daily oral treatment at 20 mg/kg, LANCL2 activation significantly reduced inflammatory immune cells including neutrophils, Th17 and Th1 cells in 3xTg mice relative to vehicle control. Further, treatment with LANCL2 agonists reduced the expression of key markers in the brain associated with impaired cognitive function to levels observed within age-matched, negative control WT mice. Activation of LANCL2 prevents the polarization of microglia into a reactive phenotype and supports the metabolic demands of amyloid beta phagocytosis. LANCL2 drugs can combat Alzheimer’s disease progression, given that reactive microglia polarization not only diminishes microglial phagocytic capacity but also increases the production of inflammatory cytokines. The LANCL2 pathway’s transformative potential merits development as a promising therapeutic mechanism for Alzheimer's disease and diverse age-related cognitive impairment.
Impact of monocytes on the proliferation of highly activated NK-like cells (GAIA-102)
Authors: Yuan Shuyue , Harada Yui , Morodomi Yosuke , Nagata Hiroko , Yonemitsu Yoshikazu 
Published: November 2024

Abstract
[Background] There has been a growing clinical application of therapies using autologous and allogeneic natural killer (NK) cells. Researchers across the field have been developing various methods to amplify NK cells, highlighting the increasing importance of NK cell amplification techniques. We have already developed a technique to amplify highly active NK-like cells (GAIA-102) from allogeneic peripheral blood mononuclear cells (PBMCs) with several cytokines, and currently conducting three clinical trials utilizing this technology. However, it remains unclear which components of PBMCs contribute to the proliferation of NK cells, a question that is central to further advancing this field.[Method] We utilized magnetic beads to deplete various blood cell components from PBMCs to investigate the effects of the presence or absence of each component on the cultivation of GAIA-102. We then evaluated the dynamics of cell proliferation and surface marker expression changes.[Results] The removal of specific components did not impact the cytotoxicity or the surface marker profile (ITGA1+/CXCR3+/CCR5+/CCR6+) of the original GAIA-102. Interestingly, eliminating monocytes from the culture markedly decreased GAIA-102 cell proliferation, a more substantial effect than when B cells were removed.[Conclusions] Our study suggests that monocytes may play a crucial role of GAIA-102 proliferation. Unraveling the impact of monocytes on this process will be a vital direction for future research.