bluuebunny/crossref_metadata_2025_split huggingface dataset:
DUSP8 induces TGF-β-stimulated IL-9 transcription and Th9-mediated immune responses by inactivating a transcriptional repressor
Authors: Chuang Huai-Chia , Tan Tse-Hua 
Published: August 2023

Abstract
Dual-specificity phosphatase 8 (DUSP8) is a MAPK phosphatase that dephosphorylates and inactivates the kinase JNK. DUSP8 is highly expressed in T cells; however, the in vivo role of DUSP8 in T cells remains unclear. Using T-cell-specific DUSP8 conditional knockout (T-DUSP8 cKO, DUSP8 f/f; CD4-Cre) mice, mass spectrometry, and chromatin-immunoprecipitation sequencing, we found that DUSP8 stimulated interleukin-9 (IL-9) gene expression and Th9 differentiation. TGF-β stimulated DUSP8 phosphatase activity in T cells. Mechanistically, DUSP8 dephosphorylated a transcriptional repressor upon TGF-β signaling, leading to the nuclear export of this transcriptional repressor and subsequent IL-9 transcriptional activation. Reduction of IL-9 mRNA levels in T cells of T-DUSP8 cKO mice was reversed by knocking out the transcriptional repressor in double knockout mice. Furthermore, IL-9 mRNA levels were induced in T cells lacking the transcriptional repressor. In addition, T-DUSP8 cKO mice displayed a reduction of IL-9 and Th9-mediated immune responses of allergy, autoimmune responses, and anti-tumor immunity in animal models. In contrast, the reduced allergy phenotype in T-DUSP8 cKO mice was reversed by knocking out the transcriptional repressor in double knockout mice. Collectively, DUSP8 induces TGF-β-stimulated IL-9 transcription and Th9-mediated immune responses by promoting the nuclear export of a transcriptional repressor.
Identification of a novel interleukin-9 receptor complex in non-hematopoietic cells
Authors: Cheung Cheuk Lam , Kaplan Mark H 
Published: August 2023

Abstract
Beyond cell-cell contact, communication at a distance requires the secretion of cytokines that bind to specific receptors on responsive cells. Interleukin (IL)-9 is a cytokine with pleiotropic effects not only on immune cells that contribute to the progression of tumor and autoimmunity, but also on structural cells including airway and intestinal epithelial cells involved in the development of inflammatory and allergic diseases. Typically, IL-9 receptor (IL-9R) functions as a complex of two subunits – the α-chain (IL-9Rα) and a common γ (γc) chain, that is shared with other cytokine receptors. Although structural cells express IL-9Rα, they lack expression of the γc chain. Thus, it is still unclear how IL-9 transmits a signal through a single IL-9Rα chain in non-hematopoietic cells. This study aimed to explore the possibility that like the type II IL-4Rα, the IL-9Rα chain can pair with the IL-13Rα1 chain, forming a type II IL-9R and can mediate IL-9 responses in epithelial cells. We demonstrated that the IL-9Rα/IL-13Rα1 complex was detected in situ in mouse tracheal epithelial cells and human small airway epithelial cells (SAEC) using proximity ligation assay. STAT3 was activated after exposure to IL-9 in vitro in SAEC when co-cultured with neutralizing antibody targeting γc chain but STAT3 activity returned to its basal level in the presence of anti-IL-13Rα1 antibody, providing further evidence of a type II IL-9R on epithelial cells. Strikingly, asthma-associated genes are upregulated in SAEC upon IL-9 stimulation in RNA-seq analysis. Collectively, our results identify a putative receptor complex - a type II IL-9R composed of IL-9Rα and IL-13Rα1, providing a mechanism for responses of structural cells to IL-9 and a potential role in asthma.
IL-2 rescues impaired T cell function upon loss of Itk via metabolic reprogramming
Authors: Kaul Zenia , Gomez-Rodriguez Julio , Golec Dominic P , Reilley Julie , Schwartzberg Pamela L 
Published: August 2023

Abstract
IL-2 inducible T cell kinase (Itk) is a critical component of T cell receptor (TCR) signaling required for activation of PLCg and downstream transcription factors, that regulate expression of effector molecules. Loss of Itk impairs T-cell activation by modulating the strength of TCR signal. Itk−/− mice have defects in T cell development with decreased proliferation and impaired differentiation to multiple CD4 T helper lineages. Most severe defects are seen in differentiation of Th9 cells, which produce the cytokine, IL-9. Under Th9 differentiation conditions, Itk−/− cells initially appear similar to WT cells but at 72hrs fail to sustain long-term expression of effector molecules. Th9 differentiation could be rescued by inclusion of IL-2. Itk−/− CD4 T cells show decreased mTORC1 signaling and do not maintain expression of IRF4, c-Myc, pRb and multiple nutrient transporters, including the Glucose transporter 1 (GLUT1), transferrin receptor (CD71), and amino acid co-transporter (CD98), all of which were rescued by IL-2. IL-2 is not required initially but can be added as late as 48hrs into culture. Using SCENITH—a flow cytometry-based technique that profiles metabolism—we found that WT differentiated Th9 cells were primarily glycolytic. In contrast, Itk−/− Th9 cells were less synthetic and more dependent on mitochondrial respiration. Inclusion of IL-2 in Itk−/− cultures rescued metabolic profiles, comparable to WT cells. Our results suggest that metabolic flexibility is central to dictating the functional/effector fate of CD4 T cells and that the Tec kinase Itk and IL-2-mediated pathways are required for maintenance of metabolic integrity, providing novel insight into mechanisms by which IL-2 can rescue T cell functional defects.
This work was supported by the Intramural Research Program of NIAID, NIH.
Quantitation of low abundant cytokines in human blood samples using SMC® high sensitivity immunoassays
Authors: Hwang Joseph B , Walden Zara , Banerjee Munmun , Gilliam Brooke , Xiao Qiang 
Published: August 2023

Abstract
Quantitation of biomarkers in blood is crucial for the understanding of inflammatory diseases, which can lead to exploration into new therapeutic applications. Although the mechanism of disease pathogenesis may be unknown, pro-inflammatory cytokines (IL-1, IL-6, IL-17, IL-18, IL-23, IFN-γ, and TNFα) and anti-inflammatory cytokines (IL-4, IL-10, and IL-13) have been implicated in the pathogenesis of a variety of diseases. In basic research, standard immunoassays, such as an enzyme-linked immunosorbent assay (ELISA), have provided valuable understanding of both normal and pathological processes for many decades. However, in more recent decades, new immunoassay technologies have uncovered numerous biomarkers, such as cytokines, in blood that were once undetectable using traditional ELISAs. In this study, we have developed high-sensitivity immunoassays, using Single Molecule Counting (SMC®) technology, to meet this new challenge of quantifying low abundant proteins in blood. The SMC® IL-4, IL-17A and IL-23 assays consistently demonstrate LLoQ of 0.04 pg/mL, 0.03 pg/mL, and 0.1 pg/mL, respectively. More importantly, these assays were able to accurately quantify cytokines in healthy and patient samples, which were once not quantifiable using traditional ELISAs. Most notably, the dynamic range of these ultrasensitive SMC® kits allowed quantitation of both healthy and disease blood samples in a single assay. In summary, SMC® high sensitivity cytokine immunoassay kits can provide a powerful non-invasive biomarker tool in studying inflammatory diseases.
Human CCR6+ Th cells show both an extended stable gradient of Th17 activity and imprinted plasticity
Authors: Singh Satya , Parween Farhat , Edara Nithin , Zhang Hongwei H , Chen Jinguo , Otaizo-Carrasquero Francisco , Cheng Debby , Oppenheim Nicole A , Ransier Amy , Zhu Wenjun , Shamsaddini Amirhossein , Gardina Paul J , Darko Samuel W , Singh Tej Pratap , Douek Daniel C , Myers Timothy G , Farber Joshua M 
Published: August 2023

Abstract
Th17 and related (type 17) cells have been investigated in mice for their contributions to autoimmune diseases and have demonstrated roles in immune-mediated disorders and host defense in humans. However, the pathways of differentiation of type 17 cells and the structure and heterogeneity of the type 17 memory population in humans are not well understood; such understanding is critical for manipulating these cells in vivo. By exploiting differences in levels of surface CCR6 and analyzing cells in bulk and as single cells, we found that human type 17 memory cells, including individual T cell clonotypes, form an elongated continuum of type 17 character along which cells can be driven by increasing the signature regulator, RORγt. On this continuum, cells are marked by extensive but progressively diminishing co-expression of genes of alternative lineages. Single-cell analysis of cytokine gene expression also revealed that Th cells initially undergo pan-activation of effector pathways rather than choose one lineage, and that these pan-activated cells persist in the memory population with multi-lineage character - reflecting both an efficient mechanism for jump starting an initial immune response and the availability of multi-potential cells in case of a repeat challenge. Along the type 17 continuum, the cells’ phenotypes and epigenomes are stable across cell divisions under homeostatic-like conditions. Nonetheless, strong activation in non-polarizing and polarizing environments can induce additional functionalities by both previously imprinted and environment-induced mechanisms that contribute differentially across the continuum to yield the unusual plasticity ascribed to even highly differentiated type 17 cells.
This study was supported by the Intramural Research Program of NIAID, NIH.
Epicutaneous Staphylococcus aureus exposure triggers lung inflammation via epithelia-intrinsic IL-36R signaling
Authors: Nolan Sabrina J , Saito Yoshine , Dikeman Dustin A , Pontaza Cristina , Ahmadi Michael , Youn Christine , Zhang Jing , Archer Nathan K 
Published: August 2023

Abstract
Allergic diseases have been increasing recently and affect nearly 20% of the human population, including 200 million people with atopic dermatitis (AD), 250 million people with food allergies, and 600 million people with allergic rhinitis or asthma. The atopic march is the concept whereby having early-in-life AD and a dysfunctional skin barrier during initial allergen sensitization increases the predisposition of allergic diseases at distant epithelial sites (e.g., asthma). We have previously shown that IL-36R signaling was required to exacerbate the progression from AD-like skin inflammation to subsequent allergic lung inflammation in mice. However, whether IL-36R acts directly on the skin and/or lung epithelia to trigger the progression towards lung inflammation is not entirely clear. To address this gap in knowledge, we performed our allergic lung inflammation model in WT, IL-36R−/−, keratinocyte-specific IL-36R deficient (K14-cre×IL-36Rfl/fl), and lung epithelial-specific IL-36R deficient (Nkx2-1-cre×IL-36Rfl/fl) mice, whereby first epicutaneous Staphylococcus aureus and cockroach antigen (CrA) exposure initiates AD-like skin inflammation, which is followed by intratracheal CrA exposure to trigger subsequent allergic lung inflammation. Evaluation of skin and lung inflammation by image, histologic and flow cytometric analyses revealed that both keratinocyte- and lung epithelial-intrinsic IL-36R signaling were required for the development of lung inflammation, weight loss, and neutrophil lung infiltration despite no differences in circulating neutrophils. Collectively, our findings suggest that IL-36R signaling in skin and lung epithelia is a critical mechanism for the progression of the atopic march.
NIAMS: R01AR073665 and LEO Foundation: LF-OC-22-000953; SJN is funded by NIAMS 1T32AR074920
Dictionary of immune responses to 86 cytokines in vivo at single-cell resolution
Authors: Cui Ang , Huang Teddy , Li Shuqiang , Ma Aileen , Perez Jorge , Sander Chris , Keskin Derin , Wu Catherine , Fraenkel Ernest , Hacohen Nir 
Published: August 2023

Abstract
Cytokines mediate cell-cell communication in the immune system and represent important therapeutic targets. A myriad of studies have underscored their central role in immune function, yet we lack a global view of the cellular responses of each immune cell type to each cytokine. To address this gap we created the Immune Dictionary – a compendium of single-cell transcriptomic profiles of over 20 cell types in response to each of 86 cytokines in murine lymph nodes in vivo, representing over 1,500 cytokine-cell type combinations. A cytokine-centric view of the dictionary revealed that most cytokines induced highly cell type-specific responses. For example, the inflammatory cytokine IL-1β induced distinct gene programs in almost every cell type. A cell type-centric view of the dictionary identified multiple cytokine-driven cellular polarization states in each immune cell type, including previously uncharacterized states such as an IL-18-induced polyfunctional NK cell state. Based on the dictionary, we developed companion software, Immune Response Enrichment Analysis (IREA), for assessing cytokine activities and immune cell polarization from gene expression data, and applied it to reveal cytokine networks in tumors following immune checkpoint blockade therapy. Our dictionary generates new hypotheses for cytokine functions, illuminates pleiotropic effects of cytokines, expands our knowledge of activation states of each immune cell type, and provides a framework to deduce the roles of specific cytokines and cell-cell communication networks in any immune response.
This work was supported by the NIH Grant RM1HG006193 and an Adelson Medical Research Foundation Grant to N.H. This work was also supported by a Natural Sciences and Engineering Research Council of Canada (NSERC) Doctoral Fellowship, Whitaker Health Sciences Fund Fellowship, and Wellington and Irene Loh Fund Fellowship to A.C., and NCI Research Specialist Award (R50CA251956) to S.L.
GRK3 Promotes Ligand-mediated Endocytosis and Limits Signaling of C-C Chemokine Receptor 7 (CCR7)
Authors: Sanchez Anahi , Bassuk Daniel W , Bill Colin A , Vines Charlotte M 
Published: August 2023

Abstract
C-C chemokine receptor 7 (CCR7) binds to two different ligands, which are differentially internalized, and mediate distinct signaling pathways. G protein coupled receptor (GPCR) kinases (GRKs) can phosphorylate ligand activated GPCRs. Therefore, we sought to determine if GRKs functioned differentially following binding of CCR7 ligands, C-C Chemokine Ligand 19 (CCL19) or CCL21 to promote distinct signaling events. Both the CRISPRCas9-modified human T-cell line, (CEM-GRK3−/−), in which both loci were disrupted, and primary murine T-cells homozygously deleted for GRK3 (T-GRK3−/−), were unable to fully internalize ligand bound CCR7. To determine if homozygous loss of GRK3, affected migration we examined chemotaxis to CCL19 and CCL21. Primary T-GRK3−/− were unable to migrate in response to CCL19 or CCL21. To examine signaling we measured mobilization of Ca2+ in primary T-GRK3−/−. In contrast to Ca2+ mobilization in CEM parental cells, which revealed sustained release of approximately 10% Ca2+ for both ligands over a 5-minute span, we found that both ligands induced a three-fold increase in Ca2+ signaling, suggesting that GRK3 plays a role in limiting the Ca2+ mobilization in T-cells to allow for migration. Overall, these observations reveal a novel role for GRKs in CCR7 signaling pathways.
Supported by grant from NIH SC1-GM111172
The Ikaros zinc finger transcription factor Aiolos as a novel regulator of T cell migration
Authors: Leonard Melissa , Read Kaitlin A , Jones Devin M , Pokhrel Srijana , Warren Robert T , Oestreich Kenneth J 
Published: August 2023

Abstract
Chemokine signaling is an integral component of lymphocyte migration, activation, and survival. The chemokine receptor CXCR3 is highly expressed on CD4+ T helper 1 (TH1) cells, which play a critical role in the adaptive immune response against intracellular pathogens. While CXCR3 helps direct TH1 cells to sites of inflammation or infection, it has also been implicated in a multitude of autoimmune diseases and cancers. Thus, understanding the molecular mechanisms that regulate CD4+ T cell migration is imperative for preventing immunopathology. Here, we identify the Ikaros zinc finger (IkZF) transcription factor Aiolos (IkZF3) as a positive regulator of CXCR3 expression in CD4+ T cells in a murine influenza model. Initially, we show decreased numbers of Aiolos-deficient antigen-specific CD4+ T cells in the lung, which correlates with decreased CXCR3 expression. RNA-seq and ATAC-seq analyses of in vitro-generated WT and Ikzf3−/− TH1 cells revealed decreased Cxcr3 transcript and reduced chromatin accessibility at regulatory regions of Cxcr3 in the absence of Aiolos, respectively. Collectively, these data imply a novel role for Aiolos in positively regulating CXCR3 expression in TH1 cells. Given these findings, we seek to define the functional effects of Aiolos deficiency on T cell migration and the molecular mechanism(s) by which Aiolos directly and/or indirectly regulates migration. Future work is aimed at determining whether Aiolos affects CXCR3 expression through direct association or indirectly via impacts on other factors.
K. J. O. is supported by a grant from The National Institutes of Health (NIH) AI134972, as well as from The Ohio State University College of Medicine and The Ohio State University Comprehensive Cancer Center.
Single-cell RNA sequencing identifies distinct transcriptomic signatures between PMA/Ionomycin and aCD3/aCD28 agonistic antibodies stimulation on primary human T cells
Authors: Lee Jung-Ho , Lee Brian H , Nam Hyo Jeong , Jeong Soyoung , Kim Hyun Je , Park Chung-Gyu 
Published: August 2023

Abstract
Immunologists have activated T cells in-vitro using various stimulation methods including PMA/Ionomycin and anti-CD3/CD28 agonistic antibodies. PMA stimulates protein kinase C, which is followed by activation of NF-κB, and Ionomycin translocates to the plasma membrane and increases intracellular calcium levels, resulting in activation of NFAT. On the other hand, anti-CD3/CD28 agonistic antibodies can activate T cells through ZAP-70, which phosphorylate LAT and SLP-76 to activate T cells. Despite the two different in vitro T cell activation methods that have been utilized for decades, the differential effects on primary human T cells following chemical-based or antibody-based activation have not been comprehensively described. As single-cell RNA sequencing (scRNA-seq) technologies enable the dissection of gene expression at single-cell resolution unbiasedly, we decided to compare the transcriptome profiles of the non-physiological and physiologic activation methods on four independent human PBMC-derived T cells. Remarkable differences in expression of cytokine and their receptor’s transcriptomes were identified. Also we identify activated CD4 T cell subsets (CD55+) enriched specifically by PMA/Ionomycin activation. We found that expression of CD40L gene was elevated on PMA/Ionomycin-treated CD4 T cells while expression of OX40 gene was elevated on aCD3/aCD28 agonistic antibody-treated CD4 T cells. We believe this human T cell transcriptome atlas derived from two different activation methods will enhance our understanding of the in vitro T cell activation assays.
Prenatally Educated Friendly NK Cells regulate Allo-specific T cell Responses through expansion of Foxp3+CD4+Treg Cells and suppression of Type 1 and 17 CD8+T cell responses
Authors: Kang Hee Kap , Papastefan Steven T , Turner Lucas , Redden Catherine , Langereis Morgan , Alhajjat Amir , Shaaban Aimen 
Published: August 2023

Abstract
Prenatal alloantigen exposure during embryonic development permanently tolerizes the early gestational fetus and provides a tremendous advantage over transplantation after birth. However, the influence of prenatally educated NK cells on alloantigen-specific T cell immune responses is not well elucidated. In previous experiments, we found that prenatally-educated friendly NK (fNK) cells exert suppressive effects on allo-specific T cell responses in vitro. We hypothesized that prenatally-educated fNK cells exert regulatory effects on allo-specific T cells in vivo.
To challenge this hypothesis, we examined the regulatory potential of prenatally educated fNK cells through gain and loss of function experiments in stable prenatal Balb/c --&gt; B6.Thy1.2+ chimeras (engrafter mice). We depleted both (Ly49AFG+) fNK cells and Thy1.2+T cells or Thy1.2+T cells alone in host engrafter mice and then adoptively transferred naïve responder Thy1.1+ T cells along with donor-specific allogeneic or syngeneic target cells into the engrafter hosts. We found that the fNK cell-replete animals exhibited a higher frequency of foxp3+Tregs and a lower frequency of allospecific T-bet+ and RORγt+ CD8 T cells compared to the fNK cell-depleted controls. Furthermore, the neutralization of TGF-β1 decreased the fNK cell-induced expansion of Treg cells in vitro.
From these studies, we conclude that prenatally educated fNK cells exert regulatory effects on allo-specific T-cell responses through the TGF-β1 mediated expansion of foxp3+Treg cells. Prenatal antigen experience drives the emergence of antigen-specific suppressors that regulate T cell responses.
National Institutes of Health Grant R01HL103745 and Lurie Children’s Hospital Research Foundation (to A.F.S.)
Zinc-c-di-AMP nanoparticles target and suppress tumors via endothelial STING activation and tumor associated macrophage reinvigoration
Authors: Yang Kaiting , Han Wenbo , Jiang Xiaomin , Piffko Andras , Bugno Jason , Han Chuanhui , Li Sirui , Liang Hua , Xu Ziwan , Zheng Wenxin , Wang Liangliang , Wang Jiaai , Huang Xiaona , Ting Jenny P , Fu Yang-Xin , Lin Wenbin , Weichselbaum Ralph R 
Published: August 2023

Abstract
Mounting evidence indicates that the stimulator of interferon genes (STING) pathway is critical for the effectiveness of standard cancer treatments. However, systemic delivery of STING agonists remains a challenge due to poor tumor-targeting and unwanted toxicities. Here we describe a robust tumor-targeted STING agonist, ZnCDA, formed by the encapsulation of bacterial-derived cyclic dimeric adenosine monophosphate (CDA) in nanoscale coordination polymers. Intravenously injected ZnCDA prolonged CDA circulation and efficiently targeted tumors, mediating robust antitumor effects in a diverse set of preclinical cancer models. Moreover, endothelial STING activation promotes the accumulation of ZnCDA in the tumor tissues by amplifying the enhanced permeability and retention (EPR) effect. In this study, we discover that type I IFN is not required for the antitumor effects of STING agonism by ZnCDA, indicating that differentially formulated STING activators may suppress tumor growth in different ways. Using metal- or dye-labelled nanoparticles, we identified a group of tumor-associated macrophages (TAMs) as the cellular target of ZnCDA in the tumor immune microenvironment. RNA-seq analysis indicated the modulation of TAMs for antigen processing and presentation via STING activation. We further showed that TAMs with ZnCDA uptake promoted an antitumor T cell response. Lastly, we utilized two immunologically “cold” tumor models, a fragment Panc02-SIY pancreatic cancer model and an intracranial GL261 glioma model, both of which represent intractable fatal human cancers, to provide a proof-of-concept strategy for combining ZnCDA with immune checkpoint blockade or radiotherapy in aggressive, difficult-to-treat tumors.
a grant from the Ludwig Foundation and an NIH grant (R01CA262508) to R.R.W., NIH grants (R01CA223184 and R01CA216436) to W.L. and NIH grants (R35CA232109 and R01AI141333) to J.P.Y.T. A.P. is the recipient of a Walter Benjamin scholarship provided by the German Research Foundation (Deutsche Forschungsgemeinschaft). J.B. was supported by a Clinical Therapeutics Training Grant (T32GM007019).