bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anti-TIGIT antibodies promote immune activation relevant to targeting stem-like and tumor-specific T cells in combination with anti-PD-1
Authors: Sivick Gauthier Kelsey E , Piovesan Dana , de Groot Amber E , Reiner Gabrielle L , Schweickert Patrick G , Soriano Ferdie , Chen Ada , Singh Hema , Zhao Xiaoning , Seitz Lisa , Reddy Anita , Young Stephen W , Walker Nigel , Walters Matthew J 
Published: January 2023

Abstract
TIGIT is an inhibitory receptor expressed primarily on NK and T cell subsets, and binding to its cognate receptor ligand, CD155, results in multiple mechanisms of immunosuppression. Blocking the TIGIT-CD155 interaction in the context of cancer promotes anti-tumor immunity. We characterized cellular subsets that TIGIT blockade may impact and the pharmacology of two anti-TIGIT antibodies - representing functional (AB308) and non-functional (AB154) Fc domain classes - undergoing clinical evaluation. Surrogate antibodies were leveraged to interrogate TIGIT biology in mouse syngeneic tumor models. Human tumor-infiltrating lymphocytes from a variety of cancer types expressed appreciable levels of TIGIT on relevant immune populations, including Tregs and CD8+ T cells with tumor reactive or pre-dysfunctional stem-like phenotypes. Both antibodies potently bound TIGIT and blocked the TIGIT-CD155 interaction as well as displayed the predicted phenotypes in terms of Fcγ receptor (FcγR) engagement. In line with FcγRIII binding, AB308 demonstrated a capacity to induce ADCC against TIGIT-expressing target cells. Combination of anti-TIGIT antibodies with other therapeutic approaches that promote T cell activation resulted in enhanced immune responses. In mice, while combining Fc-silent or Fc-enabled anti-mouse TIGIT antibody with anti-PD-1 resulted in greater tumor growth inhibition than with anti-PD-1 alone, the activity of Fc-enabled anti-TIGIT was associated with intratumoral Treg depletion. These data provide a rationale for combination with immune-activating agents and support ongoing clinical evaluation of AB154 and AB308 with biomarker strategies focused on understanding the role of Fc functionality.
Integrated immune networks in SARS-CoV-2 infected pregnant women reveal differential NK cell and unconventional T cell activation
Authors: Habel Jennifer R , Chua Brendon Y , Kedzierski Lukasz , Selva Kevin J , Damelang Timon , Haycroft Ebene R , Nguyen Thi H O , Koay Hui-Fern , Nicholson Suellen , McQuilten Hayley , Jia Xiaoxiao , Allen Lilith F , Hensen Luca , Zhang Wuji , van de Sandt Carolien E , Neil Jessica A , Amanant Fatima , Krammer Florian , Wragg Kathleen , Juno Jennifer A , Wheatley Adam K , Tan Hyon-Xhi , Pell Gabrielle , Audsley Jennifer , Reynaldi Arnold , Thevarajan Irani , Denholm Justin , Subbarao Kanta , Davenport Miles P , Hogarth Mark , Godrey Dale I , Cheng Allen C , Tong Steven Y C , Bond Katherine , Williamson Deborah A , James Fiona , Holmes Natasha E , Smibert Olivia C , Trubiano Jason A , Gordon Claire L , Chung Amy W , Whitehead Claire , Kent Stephen J , Lappas Martha , Rowntree Louise C , Kedzierska Katherine 
Published: January 2023

Abstract
Although pregnancy poses a greater risk for severe COVID-19, the underlying immunological changes associated with SARS-CoV-2 infection during pregnancy are poorly understood. We defined immune responses to SARS-CoV-2 in pregnant and non-pregnant women during acute and convalescent COVID-19 up to 258 days post symptom onset, quantifying 217 immunological parameters. Additionally, matched maternal and cord blood were collected from COVID-19 convalescent pregnancies. Although serological responses to SARS-CoV-2 were similar in pregnant and non-pregnant women, cellular immune analyses revealed marked differences in key NK cell and unconventional T cell responses during COVID-19 in pregnant women. While NK cells, γδ T cells and MAIT cells displayed pre-activated phenotypes in healthy pregnant women when compared to non-pregnant age-matched women, activation profiles of these pre-activated NK and unconventional T cells remained unchanged at acute and convalescent COVID-19 in pregnancy. Conversely, activation dynamics of NK and unconventional T cells were prototypical in non-pregnant women in COVID-19. In contrast, activation of αβ CD4+ and CD8+ T cells, T follicular helper cells and antibody-secreting cells was similar in pregnant and non-pregnant women with COVID-19. Elevated levels of IL-1β, IFN-γ, IL-8, IL-18 and IL-33 were also found in pregnant women in their healthy state, and these cytokine levels remained elevated during acute and convalescent COVID-19. Collectively, our study provides key insight to innate T cell and NK cell perturbations occurring in pregnant women with COVID-19, which will potentially inform patient management and education for those with COVID-19 during pregnancy.
Supported by National Health and Medical Research Council, Australia
Role of innate lymphoid cells on experimental sepsis-induced cognitive decline.
Authors: Giridharan Vijayasree Vayalanellore , Tripathi Ashutosh , Madeshiya Amit , Pillai Anilkumar , Petronilho Fabricia , Dal-Pizzol Felipe , Barichello Tatiana 
Published: January 2023

Abstract

Background
Sepsis is a disordered host immune reaction to pathogen invasion that leads to organ dysfunction, high mortality, and long-term consequences. More than 50% of sepsis survivors often suffer from severe and long-term cognitive deficits. We previously demonstrated in vivo microglial activation and neuroinflammation associated with cognitive deficits in an experimental model of sepsis. Recent studies have demonstrated the role of innate lymphoid cells (ILCs) in aging and neurodegeneration-induced cognitive loss. ILCs are enriched at barrier surfaces and regulate inflammation and tissue homeostasis in a disease condition. Here we aimed to explore the role of ILCs subtypes on experimental sepsis-induced cognitive deficits.

Methods
Male, 50-day-old C57BL/6 mice were subjected to cecal ligation and perforation (CLP) surgery to induce sepsis or sham surgery as a control group. At 10 days after sepsis, assessment of cognition was performed using novel object recognition (NOR) task. The ILCs subtypes ILC1, ILC2, and ILC3 were evaluated in the spleen, meninges, and choroid plexus (CP) using multicolor flow cytometry.

Results
Sepsis groups demonstrated cognitive decline as measured by decreased recognition index as compared to the control group. In the periphery, the spleen from the sepsis group demonstrated increased ILC1 and ILC3 levels as compared to control groups. In brain barriers, we observed increased accumulation of ILC2 subtype in CP in the sepsis group. However, there was no difference in the meningeal accumulation of ILCs between control and sepsis groups.

Conclusion
Targeting group 2 innate lymphoid cells may provide a new strategy to combat sepsis-associated long-term cognitive deficits.
Supported by Alzheimer’s Association grant to TB - AARGDNTF-19-619645. NIH/NIA grant to TB - 1RF1AG072491-01.
Role of Macrophage accumulation activates NLRP3 inflammasome induced IL-18 pathway in EoE pathogenesis
Authors: Sekhar Yadavalli Chandra , Venkateshaiah Sathisha Upparahalli , Lokanatha Oruganti , Mishra Anil 
Published: January 2023

Abstract

Objective
Most studies implicated eosinophilic esophagitis (EoE) to the induced esophageal eosinophilia mediated inflammation and fibrosis. However, yet the initial mechanism of allergen-induced eosinophils generation is yet not understood. Our current study provides stepwise pathway operating following the interaction of allergen and antigen presenting cells to induce EoE pathogenesis.

Methods
We performed immunostaining, RT-PCR measure transcript levels and Masson trichrome staining for collagen accumulation.

Results
We show that allergen-challenge promotes induced macrophage accumulation and epithelial cells hyperplasia in esophagus. Further, we observed that NLRP3 is induced in both macrophage and epithelial cells. The NLRP3 is the source of IL-18 and we show induced levels of IL-18 in allergen challenge mice that generate tissue eosinophilia in experimental model of EoE. Further, we observed a similar induced NLRP3 in the accumulated macrophage and epithelial cells along with induced IL-18 in the esophageal biopsies of human EoE. Mechanistically, to substantiate the role of macrophages, epithelial cells, and IL-18 role in EoE pathogenesis, we experimentally induced EoE in wild type, GM-CSF−/−, IL-18−/− mice. We show a similar NLRP3 expression with no IL-18 induction in IL-18−/− mice compare to induced NLRP3 and IL-18 in wild-type and significantly reduced NLRP3 and IL-18 in GM-CSF−/− mice. The NLRP3 regulated IL-18 induction or reduction is consistent with the tissue accumulated eosinophila and promoting esophageal remodeling and fibrosis in murine model of EoE.

Conclusions
The findings show that epithelial cells and macrophage derived NLRP3 regulated IL-18 is critical in promoting EoE pathogenesis.
Supported by NIH grant R01 AI080581
Complementary target engagement and functional assays to probe NLRP3 inflammasome pathway antagonism
Authors: O’Brien Martha A , Robers Matt , Teske Kelly , Corona Cesear , Vasta Jim , Wilkinson Jennifer , Lam Ngan , Hoffman Kaitlin Dunn , Cali James , Lazar Dan 
Published: January 2023

Abstract
Inflammasomes are critical to the innate immune system and implicated in numerous chronic and acute inflammatory diseases. Having tools to screen inflammasome modulatory drugs is critical, especially for the NLRP3 inflammasome that is triggered by a wide array of stimuli. We have developed three cell-based methods for quantifying inflammasome function: a NanoBRET™ NLRP3-specific target engagement assay, a caspase-1 activity assay and an IL-1β release assay. For the NLRP3 target engagement assay, cells are transfected with a NLRP3-Nanoluc construct and incubated with a cell-permeable, fluorescent NLRP3 binder (tracer), generating a bioluminescent resonance energy transfer (BRET) signal. The homogeneous caspase-1 and IL-1β assays can be run directly with cells in culture or with transferred culture medium, enabling application of both luminescent assays to the same cell wells via split-sample analysis. Using all three assays, we tested a series of reported NLRP3 inhibitors, including MCC950, oridonin, NBC6, NBC19, CY09, OXSi2 and OLT1177™ (dapansutrile). MCC950 inhibition of caspase-1 activity and IL-1β release was demonstrated in human and mouse cell models. We used LPS-primed THP-1 cells treated with nigericin for testing the series of inhibitors. The NLRP3 target engagement assay used HEK293 cells in suspension in non-binding plates. All three assays demonstrated good inhibition for MCC950, oridonin, NBC6, NBC19 and CY09 with rank order potency consistent with published results. We did not detect inhibition with OxSi2 or OLT1177™. These three assays enable screening for inflammasome antagonists and provide a mechanistic platform for characterizing NLRP3 inflammasome inhibitors.
CD28 signaling strength regulates the cell fate of TCF-1+ PD-1+ CD8 T cells
Authors: Humblin Etienne , Van der Heide Verena , Filipescu Dan , Lu Ashley , Selvan Myvizhi , Gumus Zeynep , Bernstein Emily , Chipuk Jerry , Homann Dirk , Kamphorst Alice O 
Published: January 2023

Abstract
During chronic infections and cancer, CD8 T cells that recognize pathogens/tumors can persist in a dysfunctional state characterized by high expression of the inhibitory receptor Programmed Cell Death (PD)-1. T cell factor (TCF)-1+ PD-1+ progenitor exhausted cells (Tpex) maintain the antigen-specific T cell pool by self-renewal and differentiation into effector-like TCF-1neg PD-1+ CD8 T cells. Previous studies described that among PD-1+ CD8 T cells, only Tpex respond to PD-1 blockade therapy. However, the molecular mechanisms that support Tpex self-renewal or differentiation are not understood. Tpex have high CD28 expression and we demonstrated that CD28 costimulation is required for response to PD-1 targeted therapy. To better understand the role of CD28 on Tpex, we evaluated how reduction or abrogation of CD28 signaling impacted virus-specific CD8 T cells in mice infected with lymphocytic choriomeningitis virus (LCMV). During established chronic infection, deletion of both Cd28 alleles resulted in reduction of Tpex and TCF-1neg subsets. In contrast, when CD28 signaling was reduced (deletion of one allele), TCF-1neg PD-1+ CD8 T cells decreased, but the number of Tpex was not affected. RNA sequencing and metabolic assays revealed that sustained CD28 signaling during persistent antigen stimulation is required to maintain mitochondrial fitness of Tpex. Our work supports the novel hypothesis that CD28 signaling strength modulates Tpex fate decision through metabolic regulation. Our data also suggest continuous CD28 signaling is required for long-term maintenance of antigen-specific PD-1+ CD8 T cells. These findings have important implications for checkpoint therapy in cancer.
Newly identified role for the transcription factor Foxp1 in natural killer cells
Authors: Canaday Laura M , Koty Amos , del Valle Kamila Andino , Cai Mingjun , Seelamneni Harsha , Krishnamurthy Durga , Waggoner Stephen 
Published: January 2023

Abstract
Natural killer (NK) cells are innate lymphocytes critical for defense against pathogenic microbes and tumors. To understand dysfunction of NK cells in disease and to facilitate effective use of NK cells as therapeutic agents in the clinic, there is a critical need to ascertain the molecular mechanisms that regulate acquisition of optimum functionality of these cells. The functional maturation of NK cells is controlled by an incompletely defined network of transcription factors, which was recently expanded to implicate numerous factors, including Foxp1, that have never been studied in these cells. We show that mice with conditional deletion of the transcription factor Foxp1 in the NK cell lineage have substantially reduced frequencies of NK cells. The remaining NK cells in these mice exhibit an immature phenotype with reduced capacity to kill tumor target cells. The identification of a role of Foxp1 in development of highly functional NK cells is significant for the development of new strategies to promote or engineer highly functional NK cells to treat patients with tumors or infections.
Supported by R01 AI148080
Defining the transcriptional regulation of NR2F6 in CD4+ T helper cell responses
Authors: Wilson Adrianna N , Campbell Sean , Lu Qun , Mosure Sarah A , He Yuanjun , Bdiri Bilel , Kamenecka Theodore M , Solt Laura A 
Published: January 2023

Abstract
NR2F6, an orphan member of the nuclear receptor superfamily of ligand-regulated transcription factors, plays an important role in CD4+ T cell differentiation and effector function. As a transcriptional repressor, NR2F6 can directly repress expression of the pro-inflammatory cytokines IL-2, IL-17A, IL-21, and IFNγ. Our overexpression data is consistent with this, demonstrating NR2F6 represses IL-17A production in TH17 cells, indicating it also may play a role in autoimmune regulation. Furthermore, genetic experiments have demonstrated the ability of NR2F6 to reduce tumor burden and develop host-protective immunological memory, a consequence of it acting as an immune checkpoint in effector T cells. Collectively, these data suggest that modulation of NR2F6 activity may have important clinical applications for autoimmune and cancer immune therapy. To date, no synthetic or endogenous ligands have been identified that modulate its activity and little is known about its transcriptional function at the molecular level. Here through integrating NR2F6 overexpression and cytokine profiling studies with -omics based approaches, such as RNAseq using T cell specific NR2F6 knockouts, we are working to elucidate NR2F6’s transcriptional function in T cells. Additionally, efforts to identify NR2F6 selective small molecules is underway so we can address how ligand modulation of NR2F6 alters its function. These experiments will help define the molecular mechanisms of NR2F6, which are poorly understood, and determine its niche in cancer immunotherapy and autoimmunity.
Supported by R01 CA225890
IL-35 suppresses pathogenic T cell response in atopic dermatitis by inhibiting PSAT1 activation
Authors: Lim Hyo Jung , Lee Jong Yeong , Choi Jin kyeong 
Published: January 2023

Abstract
Interleukin 35 (IL-35) is a member of the IL-12 family and immune-suppressive cytokine and a critical regulator of immunity during autoimmune and infectious diseases. However, it is unclear whether IL-35 can control skin inflammation by regulating the biosynthesis of biomolecules. Here, we demonstrated that IL-35 ameliorated atopic dermatitis (AD), a Th2-mediated inflammatory disease, by inhibiting activated phosphoserine aminotransferase 1 (PSAT1). We DNCB/DFE-induced AD mouse model, monitored progression by ear thickness and histology, and investigated whether IL-35 treatment would suppress AD. Mice treated with IL-35 developed mild or low skin inflammation and disease protection correlated with expansion of Foxp3+ Treg cells with concomitant suppression of Th2 and Th1/2 responses. We further show that IL-35 regulates alterations in glycolytic metabolism in AD skin inflammation. Importantly, the IL-35 mediated suppression of Th2 and Th1/2 cells and its amelioration of AD correlated with inhibition of serine biosynthesis pathways such as PSAT1 in the skin. These results suggest that IL-35 inhibits Th2 and Th1/2 cell differentiation and proliferation by directly regulating PSAT1 immune metabolites in atopic dermatitis. Therefore, the development of targeted therapy for serine-dependent metabolic pathways via IL-35 would be an effective therapeutic strategy for AD.
Supported by 21st Century Medical Science Creative Human Resource Development Center
Ameliorative role of AhR ligands in concanavalin-induced liver injury is due to increased miR-100 expression leading to mTOR downregulation and promoter hydroxymethylation
Authors: Cannon Alkeiver S , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
The promiscuous aryl hydrocarbon receptor (AhR) has long been the target of immunological studies due to its expression on many immune cells. However, the specific mechanisms underlying the role of AhR activation in inflammatory disease has not been elucidated. We employed a murine model of immune cell-mediated liver injury, comparable to autoimmune hepatitis (AIH) in humans, by intravenously injecting 12.5 mg/kg of Concanavalin A (ConA) and treating one hour later via intraperitoneal injection with an AhR ligand: 10 μg/kg 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) or 50 μg/kg 6-formylindolo[3,2-b] carbazole (FICZ). Our studies showed that both ligands improved the clinical parameters of AIH in this model. 24 hours after challenge, liver mononuclear cells were harvested, and total RNA and DNA were isolated. We examined the underlying mechanisms by conducting small RNA-sequencing and discovered dysregulation of many miRNAs. Of note, miR-100 was significantly upregulated, and we found this molecule to contain a dioxin-responsive element (DRE) in its promoter region. Further, we conducted methylated and hydroxymethylated DNA immunoprecipitation sequencing on CD45+ cells in this model and found distinct methylation profiles between each of the groups. Specifically, we observed increased hydroxymethylation in both treatment groups as compared to the ConA+Vehicle group in the promoter of mTOR, a direct target of miR-100. These data suggest that AhR activation reduces liver injury by upregulating miR-100 and reducing mTOR activity. Additionally, hydroxymethylation of the mTOR promoter may contribute to the changes observed upon AhR activation, thus lending credence for a role of hydroxymethylation in cell differentiation.
Supported by NIH grants P01AT003961, P20GM103641, R01ES030144, R01AI129788 and R01AI123947, R01AI160896 and R01AI123947-04S1
Genome-wide CRISPR screens reveal metabolic and transcriptional regulation of BTN3A and cancer susceptibility to Vγ9Vδ2 T cell targeting
Authors: Mamedov Murad R , Vedova Shane , Freimer Jacob W , Sahu Avinash Das , Chen Peixin Amy , Ramesh Amrita , Hanspers Kristina , Nguyen Vinh Q , Adams Erin J , Marson Alexander 
Published: January 2023

Abstract
γδ T cells are potent anti-cancer effectors with potential to target tumors broadly, independent of neoantigens or HLA-background. γδ T cells can sense conserved cell stress signals prevalent in transformed cells, although the mechanisms governing how γδ T cells sense and kill stressed target cells remain poorly characterized. Vγ9Vδ2 T cells – the most abundant subset of human γδ T cells – recognize a protein complex containing butyrophilin 3A1 (BTN3A1), a ubiquitously expressed cell surface protein that is activated by phosphoantigens abundantly produced by tumor cells. Here we performed genome-wide CRISPR screens in target cancer cells to identify pathways that regulate: (1) γδ T cell activity with a functional co-culture killing screen, and (2) BTN3A1 cell surface expression in a phenotypic screen. Multilayered regulation of BTN3A1 expression was uncovered: transcriptional regulation (IRF1, CTBP1, ZNF217, RUNX1), intracellular trafficking, sialylation, oxidative phosphorylation (OXPHOS), purine metabolism, and other metabolic pathways. Consistent with these results, we found upregulated BTN3A1 on cells undergoing an energy crisis due to glucose deprivation, glycolysis inhibition, or OXPHOS inhibition. Furthermore, we discovered that this BTN3A1 upregulation was induced by activation of the AMP-activated protein kinase (AMPK). Also, CRISPR screen-derived gene expression signatures correlated with higher survival in low-grade glioma patients whose tumors had high Vγ9Vδ2 T cell infiltration. Uncovering this AMPK-dependent mechanism of metabolic stress-induced ligand activation deepens our understanding of γδ T cell stress surveillance and suggests new avenues to enhance γδ T cell anti-cancer activity.
M.R.M. is a Cancer Research Institute (CRI) Irvington Fellow supported by CRI and was funded by the Human Vaccines Project Michelson Prize for Human Immunology.
An Antibody Neutralization Determinant on Domain III and the First α-Helical Domain in the Stem-Anchor Region of Tembusu Virus Envelope Protein
Authors: Yang Baolin , Meng Runze , Feng Chonglun , Huang Jingjing , Li Qiong , Wang Xiaoyan , Zhang Dabing 
Published: January 2023

Abstract
Previous studies identified three neutralizing epitopes on domains I, II, and III of the Tembusu virus (TMUV) envelope (E). More evidence is needed to understand the molecular basis of Ab-mediated neutralization and protection against TMUV. In this study, we observed a neutralizing mAb, 6C8, that neutralized TMUV infection primarily by inhibiting cell attachment. In immunofluorescence assays, 6C8 recognized the premembrane and E proteins coexpressed in HEK-293T cells, but failed to react with premembrane or E expressed individually. Epitope mapping identified nine E protein residues positioned on BC/EF loops and F/G strands in domain III and the first α-helical domain in the stem region. Further investigation with mutant viruses showed that 6C8 pressure resulted in mutations at residues 330 of BC loop and 409 of the first α-helical domain, although 6C8 only exhibited a moderate neutralizing activity in BHK-21 cells and a weak protective activity in BALB/c mice and Shaoxing duck models. Mutations A330S and T409M conferred high- and low-level 6C8 resistance, respectively, whereas the combination of A330S and T409M mutations conferred moderate-level 6C8 resistance. As a result, a quasispecies comprising three groups of antigenic variants appeared in BHK-21 cell–derived viral stocks after repeated passages of TMUV strain Y in the presence of 6C8 treatment. Taken together, these findings have raised a concern about Ab-induced antigenic variations in vivo, and they have revealed information concerning the conformational structure of the 6C8 epitope and its role in constraint on antigenic variations. The present work contributes to a better understanding of the complexity of the TMUV immunogen.