bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolic analysis of tyrosine-kinase inhibitor effect on T cell activation
Authors: Kam Yoonseok , Winer Lisa , Rogers George W , Romero Natalia , Hynes James 
Published: January 2023

Abstract
Tyrosine kinase inhibitors have been actively investigated as potential therapies for various cancers including blood tumors. However, as T cell activation can also be mediated by various tyrosine kinases, relevant inhibitors are also now being considered as pharmacological modulators of CAR-T cells; an example being dasatinib, which has been found to modulate CAR-T activity through Lck inhibition. T cell activation and subsequent differentiation are tightly associated with cellular metabolism, with T cells showing dynamic changes in two primary metabolic activities of glycolysis and mitochondrial respiration throughout the T cell life cycle. This metabolic response begins with an immediate early increase in glycolytic activity which has been shown to be an essential and signature response to activation. In this study, this fundamental biological process is exploited using the Agilent Seahorse XF technology, illustrating how activation can be measured in real-time in response to anti-CD3/CD28 activator injection, and further, how this parameter can be used to conveniently determine the modulatory effect of kinase inhibitors on T cell activation. Consistent with previous observations that kinase inhibitors can suppress naive T cell activation in vitro, here a similar kinase dependency in re-activation of resting (pre-activated) T cells was observed. Several kinase inhibitors also were able to downregulate the glycolytic rate elevated when applied post-activation. Taken together these data suggest that real-time metabolic analysis is a valuable tool in the characterization of putative T cell modulators, including tyrosine kinase inhibitors, and has the potential to inform the design of specific CAR-T modulators.
Metabolic regulation of cytokine production in leishmania resistance and susceptibility
Authors: Bodhale Neelam Prakash , Sarkar Arup , Chattopadhyay Debprasad , Saha Bhaskar , Bhattacharyya Maitree 
Published: January 2023

Abstract
Leishmania, a protozoan parasite, ensures its survival inside its mammalian host by modulating host metabolism and immune response. Recent advances in the field of immune-metabolism have shown that the metabolic enzymes and metabolites have immune-modulatory functions. Leishmania infection of a susceptible host skews immune response towards Th2-type, enhances production of pro-parasitic cytokines IL-10 and IL-4 but reduces IFN-gamma and IL-12 production. By contrast, Leishmania infection of a resistant host sets an immune response bias towards Th1-type, results in high production of pro-inflammatory IL-12 and IFN-gamma, which activates macrophages to kill the intracellular amastigotes. To understand the role of metabolic factors in Leishmania susceptibility or resistance, we used L. donovani susceptible or resistant mouse strains BALB/c or 129/Sv, respectively. The mice were first intravenously infected with L. donovani, followed by intra-peritoneal treatment with different inhibitors of metabolic pathways. The splenocytes from the treated resistant and susceptible mouse strains were studied for in-situ cytokine response and T cell subsets by FACS. Also the mouse serum was used for cytokine estimation by ELISA. The results show that different metabolic pathways regulate cytokine production in L. donovani-infected mouse suggesting their role in resistance and susceptibility to Leishmania infection.
IL-22 Effects on Osteoblast Mineralization and Metabolic Profile
Authors: Holt Vance E , Luo Yongquan , Colbert Robert A 
Published: January 2023

Abstract
Ankylosing spondylitis (AS) is an immune-mediated inflammatory disease characterized by persistent entheseal inflammation and aberrant bone formation that can lead to vertebral fusion. The Type 17 cytokine IL-17 and IL-10 family member IL-22 are involved in pathogenesis. Moreover, IL-22 has been implicated in aberrant bone formation, but its precise role remains unclear. Mesenchymal stem cells (MSCs) differentiate into osteoblasts (OBs) and are believed to reside near entheseal tissues and contribute to the observed bone formation. They express the IL-22 receptor after commitment to the OB lineage. The objective of this study was to determine how IL-22 affects OB development and mineralization in vitro. MSCs were placed in osteogenic media and then treated with IL-22 for up to 21 days after pre-conditioning with IFNg, TNF, or IFNg+TNF (I+T) for 72 hours. Mineralization was measured with Alizarin red staining, gene expression with Nanostring or RNA-seq. IL-22 significantly increases mineralization in cells pre-conditioned with I+T, compared to I+T alone or IL-22 alone, which has no effect. There was no significant acute effect of IL-22 on expression of osteogenic genes during early osteoblast differentiation in I+T treated cells. In contrast, we found that IL-22 induces a glycolytic shift within 24 hours in pre-conditioned early osteoblasts using Seahorse analysis. Given the recently recognized role of metabolic change in osteogenesis, these results suggest a novel effect of IL-22 on osteogenesis and mineralization that may be mediated by increased glycolysis. Further studies are aimed at investigating mechanisms that facilitate these effects and linking the increased matrix deposition with the metabolic shift caused by IL-22.
Progressive Immunologic Dysregulation with Exposure to Tobacco Smoke in a Model of Chronic Obstructive Pulmonary Disease (COPD)
Authors: Wu Ching-Wen , Wang Lei , Bolton Sarah , Peake Janice L. , Pinkerton Kent E. 
Published: January 2023

Abstract
Our research objective is to examine the role of dysregulated immune responses in disease progression and changes in lung physiology in a rat model of COPD. Spontaneously hypertensive rats were exposed to tobacco smoke (TS) (90 mg/m3) or filtered air (FA) for up to 12 weeks. Necropsy timepoints of 4, 6, 8, 10 and 12 weeks were chosen to define the immunological and pathophysiological changes in the lung. Rats exposed to TS showed significant increases in macrophage, neutrophil and lymphocyte number in bronchoalveolar lavage compared to FA at all weeks examined. Proinflammatory cytokines IL-1β, IL-6 and IFN-γ in whole lung homogenates were significantly increased following exposure to TS compared to FA controls. Physiological airway obstruction due to TS exposure was noted by significant elevations in central airway resistance and tissue damping. Central airway resistance was significantly increased at 4 weeks, decreased at 6 weeks, but returned to significantly elevated levels at 10 and 12 weeks following TS exposure. Tissue damping showed a peak at 4 weeks, then a decreasing trend, although remaining significantly elevated through 12 weeks of TS exposure. Strong positive correlations were noted between proinflammatory cytokines IL-1β, IL-6, IFN-γ, TNF-α and the degree of tissue damping. These changes were associated with increases in central airway squamous epithelial metaplasia and distal airway mucous metaplasia. These findings suggest progressive airway obstruction with an abnormal inflammatory response to TS exposure in this rodent model of COPD. We conclude a dysregulated immunologic response, along with airway inflammation are strongly associated with the impairment of physiological lung function in TS-exposed rats.
GPR65, a Potential Therapeutic Target for Th17-mediated Autoimmune Diseases, is Regulated by the CREB/CRTC2 Pathway
Authors: Hernandez Jeniffer B , Cave Si , Decker Carl , Swanson Jack , Yang Qiyuan , Zheng Ye , Montminy Marc 
Published: January 2023

Abstract
The cAMP response element-binding (CREB) protein has emerged as an important regulator of immune function. We have previously shown that CREB, along with its co-activator CRTC2, modulates autoimmune disease by promoting differentiation of the pro-inflammatory T cell, Th17. Th17 cells have been linked to the development of autoimmune diseases including multiple sclerosis, inflammatory bowel disease, psoriasis, rheumatoid arthritis, and asthma. The CREB pathway is induced by a variety of inflammatory signals, growth factors, and hormones that leads to the transcription of genes with cAMP-response elements. Through RNAseq, we identified multiple genes that may be regulated by CREB in Th17 cells. Here we show that the G-Protein-Coupled Receptor (GPCR), GPR65 is highly expressed in Th17 cells and is regulated by the CREB/CRTC2 pathway. The development of GPR65 antagonists may be a novel therapeutic avenue for the treatment of Th17-mediated autoimmune diseases.
Negative regulation of Ifng expression by MLL4 through CNS-28
Authors: REN GANG , CUI KAIRONG , LIU Chengyu , Zhu Jinfang (Jeff) , ZHAO KEJI 
Published: January 2023

Abstract
Interferon-g (IFN-g) is a key cytokine produced by type 1 T helper (Th1) cells in response to viral or intracellular bacterial infections in mammals. While a number of positive regulatory elements have been well studied, no negative regulatory elements at the Ifng locus for its expression have been identified. Here we report a negative regulatory mechanism of Ifng expression by MLL4, a histone methyltransferase responsible for H3K4me1. Deletion of Mll4 resulted in loss of H3K4me1 signals at the previously unrecognized CNS-28 site and led to enhanced IFN-g expression during Th1 differentiation. GATA3 bound to the CNS-28 site and deletion of Gata3 resulted in a similar upregulation of IFN-g with Mll4 deletion. Furthermore, deletion of Mll4 compromised GATA3 binding to CNS-28 and led to a substantial reorganization of the 3D chromatin structure at the Ifng genomic locus in both naïve CD4+ T cells and Th1 cells. Thus, we conclude that MLL4 negatively regulates Ifng expression by facilitating GATA3 binding to the CNS-28 site and maintaining a special local chromatin organization.
Melatonin inhibits Th17 differentiation
Authors: Davis Gwendolyn K. , Madhur Meena S. 
Published: January 2023

Abstract
Hypertension is a leading risk factor for cardiovascular disease, which is a major cause of death worldwide. T cells are required for the development of hypertension. Specifically, we showed that T helper 17 (Th17) cells produce the pro-inflammatory cytokine, IL-17A, which leads to sustained elevations of blood pressure and renal and vascular dysfunction in experimental hypertension. T cell differentiation and trafficking is regulated by circadian clocks. High fat and high salt intake, typical of Western diets, disrupt circadian rhythms and are associated with increased risk for developing cardiovascular disease. These diets are also associated with an increase in Th17 cell differentiation. Melatonin is a major circadian hormone with anti-inflammatory and anti-oxidant actions. To determine the interplay between circadian rhythms and Th17 cell differentiation, we tested the hypothesis that in vitro treatment of Th17 cells with melatonin would inhibit IL-17A cytokine production. Splenic naïve CD4+ T cells from wild type C57Bl/6J male mice were cultured for 72 hours on anti-CD3/anti-CD28 antibody coated plates in the presence or absence of Th17-polarizing cytokines and treated with melatonin (2ng/ml; Sigma Aldrich) or vehicle (0.02% DMSO). IL-17A cytokine levels were analyzed by ELISA. In cells treated with Th17 polarizing cytokines, IL-17A cytokine production was significantly reduced by melatonin. Thus, this study highlights the involvement of circadian control of Th17 cells via the actions of melatonin to suppress Th17 differentiation and IL-17A production. Future studies will determine the effect of melatonin treatment in vivo on development of hypertension and renal/vascular dysfunction.
Opioid-induced protein coding and noncoding transcriptome of human CD8+ T cells reveals opioid receptor subclass specific responses
Authors: Mazahery Claire , Llorens Braulio , Peczak Anna , Valadkhan Saba , Levine Alan D. 
Published: January 2023

Abstract
Opioid peptides are released at sites of injury; their cognate G-protein coupled opioid receptors (OR) are expressed on immune cells. Conflicting reports attribute immunostimulatory and immunosuppressive activity to opioids. From a cohort of methadone patients and controls, we found that chronic opioid use disrupts CD8+ T cell subset balance, via decreased T Effector Memory RA+ cells. Exposure of CD8+ T cells ex vivo to a μ-OR or δ-OR agonist differentially regulates thousands of protein coding genes. Gene set enrichment analysis reveals that μ-OR more strongly regulates cellular processes, including immune response via interferon, IL-2, and mTOR signaling pathways. A striking finding is the linkage of μ-OR, but not δ-OR, signaling to the upregulation of lipid, cholesterol, and steroid biosynthesis. OR-specific gene signatures associate with different transcription factors, supporting our hypothesis that transcriptional regulation underlies cellular outcomes ascribed to individual ORs. The noncoding transcriptome of opioid exposed cells is largely unique to each OR; μ-OR regulates 53 long noncoding RNAs, while δ-OR regulates 59, with only three shared transcripts, one of which is inversely regulated. The noncoding RNA upregulated by both ORs enhances integrin expression and binding; the noncoding RNA downregulated by both ORs represses STAT signaling and apoptosis. Comparison of CD4 vs. CD8 T cell responses to ligation of all three ORs (μ, δ, κ) reveals lineage specific regulation of TNF-signaling and fatty acid metabolism in response to μ-OR, G protein and cytokine receptor signaling in response to δ-OR, and REDOX pathways in response κ-OR, indicating that opioid induced immune regulation is both cell type and OR specific.
Developmental relationships of four exhausted CD8 T cell subsets reveals underlying transcriptional and epigenetic control mechanisms
Authors: Beltra Jean-Christophe , Manne Sasikanth , Hakeem Mohamed S Abdel , Kurachi Makoto , Giles Josephine R , Chen Zeyu , Casella Valentina , Ngiow Shin , Khan Omar , Huang Yinghui Jane , Yan Patrick , Nzingha Kito , Huang Alexander C. , Wherry E. John 
Published: January 2023

Abstract
Exhausted CD8 T cells (TEX) are essential during chronic viral infections and cancer. Two TEX subpopulations including a progenitor and a more terminally exhausted subset cooperate to maintain an active immune response during antigen persistence. However, non-overlapping delineations of these populations have suggested a more complex developmental biology. Here, using the LCMV mouse model of chronic viral infection, we identify four distinct TEX subsets based on Ly108 (Slamf6) and CD69 expression revealing a novel stepwise developmental framework. We reveal the transcriptional and epigenetic control mechanisms and associated biological changes underlying each TEX subset transition. Two TCF1+ progenitors were identified along with a novel TCF1-intermediate subset that re-engaged some aspects of effector biology. This subset depended on T-bet and was re-invigorated upon PD-L1 blockade. Ultimately, Tox coordinated loss of T-bet and differentiation into a fourth, terminally exhausted subset. These data define a new developmental hierarchy of TEX and reveal distinct biological properties with direct relevance to immunotherapy. Defining the control mechanisms of this TEX subset hierarchy provides novel opportunities to manipulate TEX biology for clinical goals.
USP22 controls iNKT immunity partially through MED1-mediated suppression of target-specific histone H2A monoubiquitination
Authors: Zhang Yana , Fang Deyu 
Published: January 2023

Abstract
The development and functions of CD1d-restricted iNKT cells are critical for both innate and adaptive immunity. While the ubiquitin pathway has been shown to regulate iNKT cell development and functions, the deubiquitinase involved in this process has not been identified. Herein we found that the ubiquitin-specific peptidase 22 (USP22) is highly expressed in iNKT cells during their early developmental stage I. USP22-deficiency blocked the transition from stage I to II during iNKT cell development in a cell-intrinsic manner. In addition, genetic USP22 suppression largely diminishes IL-17-producing iNKT17 and IFN-g-producing iNKT1 differentiation but favors IL-4 producing iNKT2 polarization. In contrast, genetic USP22 suppression did not alter conventional CD4 T cell clonal expansion and differentiation. At the molecular level, USP22 interacts with the Mediator of RNA polymerase II transcription subunit 1 (MED1), a transcription co-activator involved in iNKT cell development. Interestingly, while interacting with MED1, USP22 does not function as a deubiquitinase to suppress MED1 ubiquitination for its stabilization. Instead, USP22 enhances MED1 functions for IL-2Rb and T-bet gene expression through deubiquitinating histone H2A but not H2B mono-ubiquitination. Therefore, our study revealed USP22-mediated histone H2A deubiquitination fine-tunes MED1 transcriptional activation as a previously unappreciated molecular mechanism to control iNKT development and functions.
mTORC1 inhibition promotes human Treg differentiation via privileged mRNA translation
Authors: Baos Sandra Perez , Volta Viviana , Ernlund Amanda , Gadi Abhilash , Schneider Robert J 
Published: January 2023

Abstract
Maturation of regulatory T cells (Tregs) in peripheral sites is known to require TGFβ exposure and inhibition of protein kinase mTORC1. It is well known that mTOR inhibition is associated with repression of cap-dependent mRNA translation, which represents the major mechanism for protein synthesis, leaving unanswered how Tregs carry-out essential translation for development and immune suppression activity. To answer this question, we performed genome-wide transcription and translation profiling in CD4+CD127dim/− CD25+Tregs derived from anti-CD3/CD28-activated human naïve CD4+ T cells, treated with the mTORC1 inhibitor RAD001 and/or TGFβ.
We found that TGFβ activated both Treg differentiation and immune suppression genes, while mTORC1 inhibition selectively blocked translation of most T cell mRNAs except those induced by TGFβ, including FOXP3, CTLA-4, CD101 or CD103, locking in Treg lineage commitment and immune suppression function. These canonical Treg fate-determining mRNAs were resistant to mTORC1 inhibition, an effect mediated in part by their 5′-untranslated regions through an alternate form of appears to be cap-dependent, eIF4E-independent mRNA translation.
In conclusion, TGFβ transcriptional reprogramming together with mTORC1-independent translational reprogramming enable a privileged translation mechanism by which activated CD4+ T cells become Tregs.
Targeting c-Rel O-GlcNAcylation to control hyperglycemia-dependent transcription in T lymphocytes
Authors: Ramakrishnan Parameswaran , Centore Joshua , de Jesus Tristan , Church Derek , Pokorski Jonathan 
Published: January 2023

Abstract
The NF-kappaB protein c-Rel plays a critical role in controlling regulating T cell function and T regulatory cell development. We found that hyperglycemia increases O-GlcNAcylation of c-Rel at serine residue 350 and enhances the transcription of c-Rel-induced CD28RE-dependent pro-autoimmune cytokines, interleukin-2 (IL-2), interferon gamma (IFNG) and granulocyte macrophage colony stimulating factor (GM-CSF). Our recent results show that the regulatory effect of c-Rel O-GlcNAcylation is gene dependent and it suppresses the transcription of forkhead box P3 (FOXP3) expression in T cells. Mechanistically, we found that O-GlcNAcylation enhances the DNA binding of c-Rel at CD28RE and decreases it’s binding at the FOXP3 promoter. To specifically block O-GlcNAcylated c-Rel function, we developed a peptoid, called peptoid3 that specifically binds to O-GlcNAcylated c-Rel by molecular modeling and de novo synthesis. We found that peptoid3 treatment significantly decreased T cell receptor-induced, O-GlcNAcylation-dependent expression of proautoimmune cytokines and enhanced FOXP3 expression in T cells. This study reveals c-Rel S350 O-GlcNAcylation as a novel molecular mechanism inversely regulating proautoimmune gene expression and immunosuppressive FOXP3 expression in T cells with potential therapeutic implications in conditions such as type 1 diabetes.
SELENOI is important for maintenance of phosphatidylethanolamine and glycosylphosphatidylinositol anchor protein during T cell activation and immune response
Authors: Ma Chi , Hoffmann Fukun , Hoffmann Peter 
Published: January 2023

Abstract
Selenoproteins are important for both innate and adaptive immune responses. As one of the selenoprotein members, SELENOI is characterized by its ethanolamine phosphotransferase activity in the Kennedy pathway of phosphatidylethanolamine (PE) synthesis and PE is also the donor of phosphoethanolamine moiety in glycosylphosphatidylinositol anchor protein (GPI-AP). To explore how SELENOI is involved in the immune response, we generated three mouse models with different SELENOI expression levels, including defined selenium diets, doxycycline inducible SELENOI knockdown (KD) and T cell specific SELENOI knockout (KO) mice models. We analyzed both the quantity of PE species on activated CD3 T cell membrane by LCMS and GPI-AP expression on activated T cell surface by FACS in all these three models. The SELENOI KO T cells exhibit a lower level of PE species on T cell membrane compared to WT T cells, and reduced expression of GPI-anchored proteins including CD52, CD73, CD48, CD109 and CD44 on cell surface upon activation as well. We also detected the same pattern of reduced PE species and GPI-anchor proteins on defined selenium diets and doxycycline-inducible SELENOI KD mouse models. Furthermore, the SELENOI KO T cells exhibited a reduced proliferation rate upon CD3/CD28 co-stimulation. Vaccination with LCMV peptides in SELENOI deficient mice results in less CD8+ and CD4+ LCMV specific T cells and reduced IFN-γ production, indicating an impaired immune response when SELENOI is absent. These results indicate that SELENOI is crucial for both PE and GPI-AP synthesis during T cell activation and proliferation due to the importance of PE during the activation process.
Sex bias in maternal immune activation-induced neurodevelopmental disease begins at the maternal-fetal interface
Authors: Frost Elizabeth L , Lammert Catherine R. , Johanson David M. , Zunder Eli R. , Lukens John R. 
Published: January 2023

Abstract
Systemic maternal inflammation during pregnancy is increasingly thought to be a risk factor for development of autism spectrum disorder (ASD), which is diagnosed at a rate 4-fold higher in males than in females. Administration of the viral mimic polyI:C to pregnant mice at mid-gestation leads to an ASD-like phenotype in the offspring, consisting of deficits in socialization and communication as well as repetitive behaviors. In this model of maternal immune activation (MIA), elevated production of maternal serum cytokines, specifically IL-6 and IL-17, contributes to alterations in fetal neurodevelopment. Although male and female littermates are exposed to the same maternal inflammation, we show that behavioral deficits manifest only in the male offspring, mirroring the sex bias observed in human ASD. Because the placenta is derived from fetal cells and is the first site of fetal exposure to hematogenous maternal inflammation, we hypothesized that sex-specific reactions to MIA that have deleterious impacts on fetal neurodevelopment originate there. Our preliminary findings show that MIA leads to sexually dimorphic alterations in placental pathology. To further characterize responses to MIA at the maternal-fetal interface over the course of gestation, we conducted bulk RNA-sequencing of the placenta/decidua from polyI:C- and saline-treated embryos at several time points post-injection. Sex of the embryos was determined by PCR genotyping for comparison of male and female placental transcriptomes. We find previously undescribed signatures related to myeloid cells in placentas from MIA males, as well as sex-based differences under homeostasis. This data set will shed light on the immune mechanisms that impact fetal brain health.