bluuebunny/crossref_metadata_2025_split huggingface dataset:
MINK1 kinase suppresses TH17 differentiation and autoimmune inflammation via inhibition of the TGF-β pathway
Authors: Lu Linrong 
Published: January 2023

Abstract
TH17 cells (interleukin-17-producing helper T cells) are present at the sites of tissue inflammation and associated with the induction of multiple autoimmune diseases. The differentiation of TH17 cells is instructed and critically regulated by various cytokines in the microenvironment, which drive intracellular signaling pathways and transcription profiles that determine the cell fate. Here we show that MINK1, a germinal center kinase (GCK) family kinase, plays as a negative regulator for TH17 cell differentiation. Mink1−/− mice has higher frequency of TH17 cells in vivo and developed more severe experimental autoimmune encephalomyelitis (EAE) than wild type (WT) mice. Further experiments revealed that MINK1 directly interacts with SMAD2 and phosphorylates SAMD2 at T324 residue in its α-helix 1 region, which inhibits the C-terminal phosphorylation (activation) of SMAD2. Moreover, we found that MINK1 kinase is activated by T cell receptor stimulation in a ROS (reactive oxygen species)-dependent manner, which can be compromised by ROS scavenger N-acetyl cysteine (NAC). Accordingly, NAC treatment markedly boost the in vitro induction of TH17 cells in a MINK1-dependent manner and potentiates TH17 cells generation and development of EAE in vivo. In sum, our data not only demonstrate a regulatory role of MINK1 kinase in TH17 cell differentiation, but also provide the molecular insight into a possible mechanism, by which increased dietary antioxidants supplement might represent a health risk factor for the development of autoimmune diseases through increased induction of TH17 cells.
NFAT transcription factors promote the generation of follicular helper T cells in response to acute viral infection
Authors: Martinez Gustavo J , Hu Joyce , Pereira Renata , Crampton Jordan , Togher Susan , Bild Nicholas , Crotty Shane , Rao Anjana 
Published: January 2023

Abstract
Follicular helper T (Tfh) cells provide B cell help in germinal center reactions that support class switching, somatic hypermutation, and the generation of high-affinity antibodies. Here we show that deficiency in NFAT1 and NFAT2 in CD4+ T cells leads to impaired germinal center reactions upon viral infection due to reduced Tfh cell differentiation and defective expression of proteins involved in T:B interactions and B cell help, including ICOS, PD-1, and SLAM family receptors. ChIP-seq data suggest that NFAT proteins likely directly participate in regulation of genes important for Tfh cell differentiation and function. NFAT proteins are important TCR and Ca2+-dependent regulators of T cell biology, and here we demonstrate a major positive role of NFAT family members in Tfh differentiation.
Differential regulation of TH1, TFH, and TCM gene programs by cytokine-mediated events
Authors: McDonald Paul W , Read Kaitlin A. , Baker Chandra E. , Anderson Ashlyn E. , Powell Michael D. , Ballesteros-Tato Andre , Oestreich Kenneth J 
Published: January 2023

Abstract
During the course of an immune response, multiple effector T helper cell types emerge to combat the initial infection, with subsequent emergence of long-term CD4+ T memory cells to ensure that a second exposure to the same pathogen will not cause disease. CD4+ T helper cell populations that are critical for this process include T helper 1 (TH1) and T follicular Helper (TFH) effector cells and long-lasting central memory T (TCM) cells. Previous work has demonstrated a level of flexibility between effector TH1 and TFH-like cell populations, with the respective gene programs regulated by alterations in interleukin 2 (IL-2) signaling. Here, we demonstrate that in response to decreased IL-2 signaling, effector TH1 cells are capable of initiating the expression of both TFH and TCM-like gene programs, including cell surface expression of IL-6Ra and IL-7R. Importantly, exposure of these cells to either IL-6 or IL-7 augments or inhibits the TFH gene program, respectively, suggesting a potential mechanism whereby cytokine availability may influence TFH or TCM cell function. Consistent with this model, we have identified a novel population of IL-6Ra+IL-7R+ cells that emerge at late time points post-infection in an in vivo mouse model of influenza infection. Further research into the temporal and spatial expression of IL-2, IL-6, and IL-7 in the modulation of these cells represents an exciting avenue of research with wide-ranging therapeutic implications.
Mis-acylation of tRNAs suppresses protective immune responses which control cutaneous inflammation by infection
Authors: HWANG HyunJu , Lee Byunghyuk , Lee Jeong Woong , Hong Changwan 
Published: January 2023

Abstract
The mouse sticky mutation which is a missense mutation in editing domain of the alanyl-tRNA synthetase gene impairs the proofreading activity during amino-acylation of tRNAs. An editing-defective tRNA synthetase induces the accumulation of mis-folded proteins in the neuron which is associated with neuro-degenerative disorder. Although the sticky mice were characterized by the rough, unkempt, sticky appearance, it is not clear whether the sticky mutation is associated with inflammatory disease. The protective role of T and B cells during cutaneous infection by a variety of pathogens has been known well. We therefore investigated how sticky mutation affects immune responses and linked to sticky phenotype. First of all, we examined immune cell numbers in sticky mice. LN cell number in sticky mice was significantly reduced compared to Sti/+ and WT mice, while LN profiles of sticky mice were not big different from control groups. Reduced cellularity by sticky mutant results from impaired proliferation, not by IL-7 signaling which is critical in T cell development and homeostasis. Interestingly, we found that mutation in tRNA processing reduced IFNγ and IL-17 production from stimulated T cells. And it also reduced Immunoglobulin production in B cells, particularly IgG1 and IgE which has an important role in resistance to bacterial and parasite infection. These findings indicate that disruption of translational fidelity induces deterioration of immune functions such as impaired cytokine and antibody production, resulting in reduction of the resistance to various infections. Furthermore, it provides a novel mechanism which the alteration of tRNA pool in inflammatory diseases affects the cellular and humoral immune responses.
IL-22 Increases Metastatic Properties in Androgen-Independent Prostate Cancer
Authors: Shafran Jordan , Andrieu Guillaume , Denis Gerald V 
Published: January 2023

Abstract
New evidence is emerging that suggests that a subpopulation of individuals characterized as metabolically abnormal obese (“MAO”; i.e.: co-morbid Type 2 diabetes (T2D), hypertension and systemic inflammation) have a much greater risk for cancer mortality compared to obese but metabolically healthy individuals (“MHO”; i.e.: non-diabetic, lack hypertension, non-inflamed). Because MAO individuals have systemic and local adipose inflammation associated with obesity co-morbidities, it is now hypothesized that alterations to signal transduction pathways in the tumor microenvironment (which come as a result of increased T2D-associated Th17 cytokines (IL-17A, IL-17F, IL-21, IL-22, etc.)), lead to a more advanced, pro-metastatic cancer phenotype. Furthermore, additional observations have also shown that Bromodomain and Extraterminal domain (BET) proteins (BRD2, BRD3 and BRD4) act as transcriptional effectors for these Th17 cytokines. Based on these data, we sought to determine whether (i); T2D-associated Th17 cytokines increase metastatic properties in androgen-independent prostate cancer cells, (ii) signaling pathways utilize Th17 cytokines to exacerbate metastatic properties and (iii) BET proteins act as effectors for Th17 cytokine signaling. Here we report that IL-22 increases migration and invasion in androgen-independent prostate cancer. This phenotype is achieved, in part, through the activation of STAT3 in a BRD4-dependent manner. Moreover, BRD2 opposes BRD4 in this process. Overall, these results identify a novel mechanism that is critical for prostate cancer metastasis.
Unique roles of TECs, DCs and B cells in Treg repertoire formation
Authors: Watanabe Masashi , Breen Michael , Alkhaleel Farrah , Hodes Richard J 
Published: January 2023

Abstract
Foxp3+ CD4+ regulatory T cells (Treg) are an essential component in maintenance of immunological tolerance against self. Treg are generated in the thymus by relatively strong TCR recognition of self-peptide-MHCII complex (pMHCII) on antigen presenting cells (APCs). In addition to pMHCII dependent TCR signaling, B7 expression on APC is critical for Treg generation through CD28 co-stimulation. APCs that might be capable of providing such TCR/CD28 signaling to developing thymocytes include thymic epithelial cells (TECs), dendritic cells (DCs) and B cells. To date, however, how each APC contributes to the generation of the Treg repertoire specific for self-antigen under physiological condition is not fully understood. To address this question, we generated B7 conditional knockout mice enabling selective deletion of B7 from each APC type and assessed the effect of this cell type-specific perturbation of APC function on Treg generation. We first observed that B7 deletion from DCs, but not TECs or B cells, resulted in a substantial decrease in total Treg numbers. Interestingly combined B7 deletion from two (TECs + DCs) or three (TECs + DCs + B cells) APCs showed additive effects in decreasing the total Treg population size. In contrast, we found that the pMOG-specific Treg population depended on B7 on DCs, with no individual or additive effect of B7 deletion from TECs or B cells. These results suggested that TECs, DCs and B cells have unique and non-redundant roles in Treg repertoire formation. Further studies are currently underway to determine the role of TECs, DCs and B cells for the generation of Treg repertoires with distinct self-antigen specificities by utilizing panel of pMHCII-tetramers.
Early differences in TCR signal strength modulate effector and memory differentiation of CD4+ T cells responding to viral infection.
Authors: Snook Jeremy , Williams Matthew A 
Published: January 2023

Abstract
Heterogeneity in TCR-dependent activation signals influences both effector differentiation and memory formation of CD4+ T cells. Our previous studies found that the formation of CD4+ memory T cells specific for lymphocytic choriomeningitis virus (LCMV) was associated with the selection of TCRs with sustained antigen binding properties. The goal of the current study was to define early TCR-dependent activation events that promote memory differentiation. We found that early in vitro and in vivo activation of distinct T cell clones specific for the same LCMV antigen demonstrated highly variable NFAT transcriptional activity and expression of early activation markers such as CD25. Clones with enhanced memory potential had reduced NFAT activity in vitro and rapid down-regulation of CD25 in vivo. We further observed decreased CD3ζ and Zap-70 phosphorylation, as well as a decrease in NFAT-dependent gene transcription, in virus-specific CD4+ T cells that displayed decreased CD25 surface expression early after LCMV infection. Adoptive transfer of CD25hi or CD25lo effector T cells (d3 or d5 post-infection) into infection-matched hosts revealed that the vast majority of both Th1-like and Tfh-like memory cells derived from CD25lo effector cells. RNASeq analysis further confirmed enrichment of genes associated with enhanced memory development within CD25lo effector cells, including Tcf-1 and OCA-B, but decreased expression of genes associated with enhanced effector activity. We conclude that differences in TCR signal strength account for clonal differences in CD4+ memory T cell differentiation, with weaker early TCR signals permitting long-term memory formation without terminal effector differentiation.
Human memory CD8 T cell effector potential is epigenetically preserved during in vivo homeostasis
Authors: Abdelsamed Hossam Aly 
Published: January 2023

Abstract
Maintenance of memory CD8 T cell quantity and quality through antigen-independent homeostatic proliferation is vital for sustaining long-lived T cell-mediated immunity, yet the underlying mechanisms that preserve memory T cell functions during homeostasis remain largely unexplored. Here we show that preservation of effector-potential among human memory CD8 T cells during in vitro and in vivo homeostasis is coupled to maintenance of memory-associated DNA methylation programs. Whole-genome bisulfite sequencing of primary human naïve, short-lived effector memory (Tem), and longer-lived central memory (Tcm) and stem cell memory (Tscm) CD8 T cells identified demethylated promoters of effector molecules that are poised for rapid expression among all memory cell subsets. Effector-loci demethylation was heritably preserved during IL-7 and IL-15 mediated in vitro cell proliferation. In contrast to the effector-potential, antigen-independent proliferation induced a phenotypic conversion of Tcm and Tscm memory cells into Tem cells that was coupled to increased methylation of the CCR7 locus. Furthermore, in vivo proliferation of haploidentical donor memory CD8 T cells in lymhodepleted recipients resulted in a similar preservation of effector-associated methylation programs while enriching for Tem-associated programs. These data demonstrate that long-lived human memory CD8 T cells retain the ability to undergo antigen-independent epigenetic reprogramming during their developmental conversion into other memory subsets while at the same time preserving the poised effector state utilized by all memory T cells.
Defining the role of CD69 in the formation of resident memory CD8+ T cells
Authors: Walsh Daniel , da Silva Henrique Borges , Hamilton Sara , Masopust David , Jameson Stephen 
Published: January 2023

Abstract
Resident memory CD8+ T cells (TRM) occupy parenchymal tissues and have a well-defined role in protecting from reinfection. CD69 is often considered a definitive marker of TRM, and CD69 can regulate T cell trafficking through its interaction with the G-protein-coupled receptor S1PR1. However, the functional requirement for CD69 in controlling the generation and maintenance of CD8+ TRM is unclear. We tested this using co-transfer of CD69 sufficient and deficient P14 CD8+ T cells specific for gp33/Db in the context of acute LCMV and Flu-GP33 infections. Surprisingly, we found that CD69 was not necessary for TRM establishment in most tissues. However, we found that depending on the infectious agent, CD69 could promote TRM localization in certain sites. We propose that the requirement for CD69 is context dependent, and that a combination of factors including the tissue microenvironment and infectious agent dictate its influence on development of resident memory.
ATP receptor P2rX7 direct the generation of long-lived memory CD8+ T cells
Authors: da Silva Henrique Borges , Wang Haiguang , Fonseca Raissa , Block Katharine E. , Masopust Davd , Hogquist Kristin Ann , Jameson Stephen C. 
Published: January 2023

Abstract
P2rX7 is an ATP-sensitive purinergic receptor often known for its role in neuropathic pain and innate immunity. P2rX7 is also known to mediate CD4+ T cell death upon stimulation. To evaluate whether CD8+ T cell immune responses are also regulated by P2rX7, we infected mice with LCMV and tracked gp33-specific P14 CD8+ T cells. Unexpectedly, we found that P2rX7 expression on memory precursors (MPEC) positively correlated with generation of central memory CD8+ T cells (TCM). P2rX7KO CD8+ T cells also had reduced memory generation, especially TCM and resident memory (TRM). This defect was cell-intrinsic, as revealed by bone marrow chimeras and co-adoptive transfer of P2rX7KO and WT P14 T cells. Defective memory generation by P2rX7KO CD8+ T cells was preceded by impaired MPEC generation as well as diminished Tcf1 expression. Strikingly, P2rX7KO CD8+ T cells had an early and sustained impairment of proper mitochondrial function, as observed by flow cytometry (lower membrane potential and mitochondrial mass) or seahorse assays (lower SRC, higher proton leak). These defects were observed specifically on TCM, but not on effector memory (TEM) or long-lived effector cells (LLEC). P2rX7KO CD8+ T cells also showed reduced recall proliferation following secondary antigen stimulation. Importantly, treatment with P2rX7 pharmacological inhibitors during the immune response effector phase lead to similar defects in TCM and TRM. These findings show that P2rX7 is critical for development of a long-lived memory CD8+ T cell response. Also, they have important implications for the immunological effects of P2rX7 inhibitors used for pain control, and the impact of loss-of-function P2rX7 polymorphisms on immunity and the response to vaccines.
ADAMs10/17 deficient mast cells exhibit decreased IgE-mediated activation in vitro.
Authors: Luker Andrea , Conrad Daniel 
Published: January 2023

Abstract
Allergies and asthma are common ailments that are on the rise around the world. Mast cells play a direct role in the signs and symptoms characteristic in allergic patients. The family of A Disintegrin And Metalloproteinases (ADAMs) are involved in regulating many cellular processes by cleaving surface receptors, ligands, and signaling molecules. We sought to determine the role of ADAMs10/17 in bone marrow-derived mast cell (BMMC) activity. In dual inhibitor-based studies, percent degranulation and cytokines released by IgE-mediated activation were significantly reduced. Interestingly, ionomycin-activation was unchanged with inhibitor treatment, suggesting that ADAMs10/17 may be involved in IgE-mediated mast cell activation upstream of calcium influx. Preliminary results with ADAM17MC−/−BMMCs have confirmed the inhibitor data and provided interesting results regarding increased mast cell survival and proliferation. These data suggest that targeting ADAMs10/17 could attenuate mast cell-related TH2 responses.
Mechanistic insight into the suppressive effects of stem cell factor during IgE-mediated activation in bone marrow-derived mast cells.
Authors: Lyons David O , Pullen Nicholas A 
Published: January 2023

Abstract
Treatment of bone marrow derived mast cells (BMMCs) with stem cell factor (SCF) is common practice when differentiating mast cells for experimentation. The enhanced growing potential from including SCF during mast cell culture and differentiation allows for easier maintenance and quicker experiments. Recent publications have begun to identify potentially negative effects of treatment with SCF regarding mast cell activation. The mechanism of SCF inhibition on mast cell activation is mostly unknown, but BMMCs derived with the addition of SCF function differently than untreated cells when mast cells are primed and subsequently activated. This study provides insight into the specific targets of SCF treatment and the impact SCF has on the ability of mast cells to activate. Running ELISAs in conjunction with real-time qPCR, we identified differentially expressed proteins involved in FCɛRI signaling and mast cell activation in BMMCs exposed to SCF. We examine the effects of SCF on Fyn, Lyn, Syk and Stat5 protein expression compared to mast cells raised without SCF. These data are also placed in the context of other immunosuppressive mechanism, (e.g. TGFβ). The presented data allow researchers to better understand the implications that may result from SCF treatment of BMMCs, especially with respect to examining IgE-mediated functions.
Suppression of mast cell activation by exopolysaccharide produced by Bacillus subtilis.
Authors: Swartzendruber Julie A , Tolia Sanika , Knight Katherine L 
Published: January 2023

Abstract
The incidence of allergies is on the rise, and to prevent or treat allergic diseases, we need a means of inhibiting immune pathways leading to allergic responses. Reduced exposure to microbes, increased use of antibiotics and changes in diet all lead to an increase in allergies. The consequences of these changes in lifestyle include an alteration of intestinal bacteria that are important for development of the immune system. Our work seeks to utilize the beneficial effects of gut microbiota to manipulate allergic responses. A key cell responsible for the symptoms of allergies is the mast cell. We are using the probiotic bacterium Bacillus subtilis which produces an exopolysaccharide (EPS) with anti-inflammatory activity. We found that EPS treatment alone was unable to alter mast cell degranulation following IgE-mediated activation, however treatment with peritoneal cells from EPS-injected mice prevented mast cell degranulation. We hypothesize that EPS induces a suppressive cell in the peritoneal cavity that leads to the suppression of mast cell activation and could be used to suppress the immune response against common food or environmental factors.
The E3 ligase Itch suppresses the development of pathogenic Basophils
Authors: Rathinam Chozha Vendan , Thummar Keyur 
Published: January 2023

Abstract
Basophils are one of the major players in the pathophysiology of allergy related immune disorders. In spite of their key roles in autoimmune and inflammatory disorders in humans, molecular mechanisms that control basophils remain largely unknown. Therefore we have been involved in elucidating the biochemical and molecular mechanisms through which development and functions of basophils are controlled in the immune system. In this study, we utilized the Itch deficient mouse as a model system for understanding the biology of Basophils. Itch is an E3 ubiquitin ligase that belongs to the HECT family. Mice deficient for itch develop severe autoimmune defects (‘itchy’ phenotype) and patients with ITCH mutations show characteristic clinical features, including multisystem autoimmune disease. Our preliminary analysis indicated that itch deficiency leads to Basophilia, hyperactivation of basophils, elevated IL4 expression and Th2 priming by basophils, in a cell intrinsic manner. Strikingly, depleting basophils in itch deficient mice resulted in rescue of the ‘itchy’ phenotype. Our mechanistic studies demonstrated that notch signals are elevated in itch deficient basophils, due to reduced ubiquitylation of notch. Interestingly, ablation of notch signals in itch deficient mice resulted in normal frequencies of basophils and rescue of the ‘itchy’ phenotype. At this annual meeting, we would be discussing the precise involvement of the signaling pathways and molecular mechanisms in the restriction of pathogenic basophils.