bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of Interferon Genes is Influenced by Sex Hormones in SLE
Authors: Singh Ram Pyare , Hahn Bevra H 
Published: January 2023

Abstract
Environmental factors, genetic defects, miRNA dysregulation, and hormonal factors can influence susceptibility to SLE. Estradiol has been shown to increase the expression of interferon genes and pathways and thus could promote susceptibility to the disease in women. Recent evidence suggests the existence of a nexus between inflammatory pathways and 17b-estradiol, resulting in increased interferon stimulated genes (ISGs), autoantibodies and dysregulation of immune cells in SLE. The mechanisms by which 17b-estradiol interacts with IFN and various immune cells in SLE is poorly understood.
The goal of this study is to determine the role of estradiol in the regulation of candidate target genes, miRNAs, and mechanisms of regulation of autoantibody production in SLE. Women may be more susceptible than men to SLE and other autoimmune diseases in part because many healthy women have higher interferon-regulated genes. We found that plasma estradiol levels positively correlated with levels of serum/plasma IL-6 and 21/IL-23 in SLE patients. Plasma 17b-estradiol levels are significantly increased in female SLE patients compared to healthy females (p&lt;0.01). Furthermore, we found that estradiol increases pro-inflammatory cytokines, and level of estradiol positively correlated with expression of miR21, 25, and 186 in SLE patients. Expression of miR21, 25, and 186 are positively correlated with SLEDAI score in SLE.
We also found that expression of interferon genes (IL-12, 17, 21) is influenced by sex hormones in SLE. Understanding the role of 17b-estradiol in the regulation of pro-inflammatory pathways, ISGs, and microRNAs will further advance our understanding of SLE disease pathology and may identify additional novel therapeutic targets.
Characterization and Optimization of SLE Mouse Models for the Evaluation of Immunomodulation Therapeutics
Authors: Case Laure , Knowlton Hannah , Christianson Greg , Vann Lewis , Lutz Cathleen 
Published: January 2023

Abstract
Systemic Lupus Erythematosus is a chronic autoimmune disease that causes systemic damage throughout the body and is characterized by a broad spectrum of clinical manifestations. The pathogenic mechanisms underlying the disease remain unclear, but are thought to be the result of gene-by-environment interactions that lead to the breakdown of self-tolerance in susceptible individuals. Mouse models of SLE have been instrumental in our understanding of the genetic and cellular requirements for SLE induction, and represent a valuable resource for testing SLE therapies and drug candidates in vivo. No one mouse model completely recapitulates the complexity of the human disease, thus it is crucial to evaluate the mode of action and therapeutic efficacy of targeted therapeutics in multiple SLE mouse models. Here we demonstrate an effective program for screening therapeutic candidates using two well-established SLE mouse models, NZBWF1/J mice and MRL/MpJ-Faslpr/J mice, which mimic the genetic and pathophysiological aspects of the human disease. We characterize the disease course in each model in order to optimize the treatment schedule for either prophylactic or therapeutics drug efficacy studies. Using the immunosuppressant drug cyclophosphamide, we longitudinally compare the clinical measures of the disease to immune cell profiles during both in-life and at study terminus. This program provides a valuable resource for the investigation into the specific effects novel therapeutics have on immune homeostasis.
MAP4K3/GLK-induced AhR-RORγt complex is a novel therapeutic target for GLKhighIL-17Ahigh subpopulation of SLE patients
Authors: Chuang Huai-Chia , Tan Tse-Hua 
Published: January 2023

Abstract
The cytokine IL-17A plays critical roles in the pathogenesis of autoimmune diseases. The frequencies of MAP4K3 (also named GLK)-overexpressing T cells are correlated with disease severity of systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and adult-onset Still’s disease. T-cell specific GLK transgenic mice develop spontaneous autoimmune responses. GLK signaling selectively stimulates IL-17A production in murine T cells through inducing AhR-RORγt complex formation. Here, we investigated whether GLK-induced AhR-RORγt complex in T cell is a therapeutic target for human SLE or RA. The population of GLK+IL-17A+ T cells was enhanced in the peripheral blood from SLE patients compared to that of healthy controls using flow cytometry. The ROC curve analyses showed that increased GLK+IL-17A+ T cell population in peripheral blood reflected an active stage of SLE. In addition, peripheral blood T cells from SLE or RA patients displayed induction of RORγt phosphorylation and the AhR-RORγt complex. Moreover, we identified a small-molecule GLK inhibitor that inhibited GLK kinase activity and AhR-RORγt interaction. The GLK inhibitor suppressed the disease severity in autoimmune mouse models and IL-17A production in T cells from SLE or RA patients. Collectively, GLK-induced AhR-RORγt complex is a therapeutic target for the GLKhighIL-17Ahigh subpopulation of SLE or RA patients.
Development of citrullinated-vimentin-specific CAR for targeting Tregs to treat autoimmune rheumatoid arthritis
Authors: Raffin Caroline , Muller Yannick , Barragan Jessica , Zhou Yu , Piccoli Luca , Lanzavecchia Antonio , Tareen Semih U , Fontenot Jason D , Bluestone Jeffrey A 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by an aberrant inflammation of the synovial membrane that causes irreversible joint and bone damage. Regulatory T cells (Tregs) are defective in RA patients while adoptive transfer of expanded autologous Tregs efficiently reverse disease in collagen-induced arthritis, an animal model of RA. Moreover, providing an antigen (Ag) specificity to Tregs render them more potent to suppress abnormal inflammation by increasing specific activity and promoting selective migration to the Ag-expressing tissue, reducing the risk of pan-immunosuppression in the meantime. These studies strongly suggest that Treg therapy may be effective in the treatment of RA patients by reducing joint inflammation and inducing immune tolerance and may be improved by using Ag-specific Tregs. Therefore, we aimed to engineer Ag-specific Tregs from polyclonal Tregs using a chimeric antigen receptor (CAR) specifically targeting an Ag present in the joint of RA patients. We generated a CAR directed against a post-translationally modified intermediate filament protein, citrullinated vimentin (CV), an Ag found, abundantly and almost exclusively, in the extracellular matrix of the inflamed synovial tissue of the RA patients. CV-specific CAR was transduced into human Tregs that were efficiently activated, expanding and suppressing following CAR-mediated stimulation. Importantly, CV-CAR Tregs were shown to react with CV expressed in RA patient synovial fluid. Studies are underway to demonstrate the functional activity of these CV-CAR Tregs in in vivo mouse models, a validation step in the development of a promising therapeutic tool to treat RA and potentially other autoimmune disorders.
Phenotypical and functional analysis of natural killer cells from primary human head and neck squamous cell carcinomas
Authors: Korrer Michael J , Kim Young 
Published: January 2023

Abstract
The purpose of this study was to determine if natural killer (NK) cells from primary human head and neck squamous cell carcinomas (HNSCC) expressed altered phenotypical and functional capabilities compared to matched NK cells from the periphery. NK cells are a critical part of the host defense against cancer through direct cytotoxicity of transformed cells and the release of pro-inflammatory cytokines. To facilitate the targeting of NK cells for immunotherapy we phenotyped human HNSCC NK cells using flow cytometry and RNA-SEQ analysis of matched flow-sorted PBMC and tumor infiltrating NK cells.
We found that tumor infiltrating NK cells of HNSCC patients, significantly downregulated activating receptors NKG2D, DNAM-1, NKp30, CD16 and 2B4 compared to matched blood NK cells (n=13). Surprisingly, the majority of inhibitory KIR receptors on tumor-infiltrating NK cells were decreased. However, the inhibitory receptors NKG2A and PD-1 were found to be highly overexpressed in tumor-associated NK cells. In addition, RNA-SEQ analysis of flow sorted NK cell populations from matched blood and TIL of HNSCC patients (n=9) demonstrated significant differential expression of multiple immune pathways. Functional assays confirmed that tumor infiltrating NK cells have reduced cytotoxicity and produce significantly less IFNγ after ex-vivo stimulation (n=10).
We believe that this study provides the first in-depth look at NK cells from primary HNSCC tumors. We found that tumor associated NK cells express a profoundly different phenotype than NK cells from the blood of the same patients, with multiple activating receptors downregulated and the inhibitory receptors NKG2A and PD-1 overexpressed.
Exosomal communication by metastatic osteosarcoma cells contributes to the modulation of alveolar macrophages to an M2 tumor-promoting phenotype
Authors: Wolf-Dennen Kerri K , Gordon Nancy , Kleinerman Eugenie S 
Published: January 2023

Abstract
Osteosarcoma (OS) metastasizes to the lung and there is a link between tumor-promoting M2 macrophage content and poorer patient survival. Conversely, high M1 content correlates with improved survival. M2 macrophages are immunosuppressive, contribute to tumor progression and metastasis, and can be induced by various stimuli in the tumor microenvironment, including exosomes (Exo). Exo are small 40–150 nm vesicles that are involved in inter-cellular communication, and contribute to tumor progression, increased tumor survival and immune evasion. Because of the link between M2 content and inferior patient survival, we evaluated the effect of Exo from non-metastatic K7 OS cells and the metastatic subline K7M3 on macrophage phenotype and function. Incubating MHS mouse alveolar macrophage cells with K7M3 Exo induced mRNA expression of IL10, TGFB2, IL1rα, and CCL22, (markers of M2 macrophages), and decreased phagocytosis, efferocytosis and macrophage-mediated tumor cell killing. In contrast, Exo from non-metastatic K7 or 3T3 normal fibroblast cells did not inhibit phagocytosis, efferocytosis or macrophage-mediated tumor cell killing. Our data suggest that the Exo from metastatic OS cells can modulate cellular signaling of tumor associated macrophages, thereby promoting the M2 phenotype and inhibiting their phagocytic and cytotoxic abilities. Identifying the factors within Exo from metastatic OS cells and the mechanisms responsible for Exo communication between metastatic OS cells and macrophages could reveal new therapeutic targets and opportunities to improve the treatment of patients with metastatic disease in the lung.
Frequency of Myeloid Derived Suppressor Cells in Spleen and Peripheral Blood from Cancer Patients
Authors: Talmadge James E , Cole Kathryn , Ly Quan , Hollingsworth Michael , Cox Jesse 
Published: January 2023

Abstract
The peripheral blood (PB) of cancer patients is frequently neutrophilic and lymphopenic with few studies focused on tissue immunity. In this study we examined eight paired sets of PB and spleen populations to assess the frequency and sub-populations of myeloid derived suppressor- (MDSC) and T-cells. These studies observed a significant decrease in the frequency of granulocytic (G), monocytic (M) and immature MDSC’s in the spleen vs PB (4.8±1.6 vs. 54.4±6.1; 0.2±0.1 vs. 0.9±0.4; and 0.6±0.1 vs. 2.3±0.8 % respectively) but not T-regulatory cells (0.8±0.5 vs. 0.1±0.04 percent). The difference in the G vs M MDSC was consistent with arginase and nitric oxide synthase expression. In contrast, the % of splenic T-cells was increased relative to the PB including CD3+ (35±5 vs. 16±4%), CD4+ (19±2 vs. 10±2%) and CD8+ (10±1 vs. 5±2%). The frequency of transitional memory T-cells (Ttm) was significantly higher in the spleen vs. PB (37.5±6 vs. 26±3.0%) as was the frequency of PD1+CD8+ T-cells (8±2 vs 1±0.5% respectively). The increase in Ttm with their heightened proliferation, effector potential and tumor specific cytotoxicity of PD1+CD8+ cells support the superiority of spleen vs PB cells for adoptive cellular therapy. In support of this both the PD1+ cells were LAG3− and TIM3−, and thus not exhausted. In summary, the spleen cells of cancer patients have an increase in tumor specific effector cells with proliferative potential supporting their utility in therapeutic protocols.
Non-small cell lung cancer tumor microenvironment induces Arginase1 in neutrophils via the Toll-like receptor 2 signaling pathway
Authors: Zhang Huajia , Kargl Julia , Zhu Xiaodong , Wang Xiaoxia , Cao Jianhong , Houghton A. McGarry 
Published: January 2023

Abstract
Neutrophils dominate the tumor microenvironment of non-small cell lung cancer. Previous studies show neutrophils suppress T cells and promote immunosuppression. The objective of the current study is to identify how tumor associated neutrophils (TAN) adopt the T-cell suppressive entity after being recruited into the tumor microenvironment.
In the murine lung squamous cell carcinoma autochthonous tumor model (AdCre-Lkb1fl/fl, Ptenfl/fl; LP) and subcutaneous tumor model (Lewis lung carcinoma cell line; LLC), we found that Arginase 1 (Arg1) mRNA level in TAN is drastically higher than neutrophils from the bone marrow, spleen and peripheral blood. Arg1 suppresses T cell through Arginine depletion. Whole tumor lysates from LP tumor and LLC tumor induce Arg1 level in bone marrow-derived and peripheral neutrophils from healthy B6 mice in vitro, suggesting that the tumor microenvironment alters neutrophil properties. To identify the putative molecule(s) that induce Arg1 in TAN, we used fast protein liquid chromatography (FPLC) and mass spectrometry to analyze the whole tumor lysates from LP tumor and LLC tumor. Annexin A1 and 2 are among the common molecules that appear in Arg1-inducing FPLC fraction in both LP and LLC tumor lysates. Annexin A1, A2 are involved in toll-like receptor signaling. Flow cytometry showed more than 70% of Arg1+ TAN are either TLR2 or TLR4 positive. Interestingly, only TLR2 but not TLR4 inhibition blocked Arg1 induction by whole tumor lysates in naïve neutrophils. Taken together, tumor microenvironment induces Arg1 level in neutrophils in a TLR2 dependent manner. TLR2 is a putative therapeutic target for blocking tumor associated neutrophil-mediated immunosuppression in non-small cell lung cancer.
Single cell RNA sequencing reveals distinct gene expression signatures of myeloidderived suppressor cells in breast cancer
Authors: Alshetaiwi Hamad , Pervolarakis Nicholas , McIntyre Laura L. , Ma Dennis , Nguyen Quy , Nee Kevin , Rath Jan , Evans Katrina , Torosian Leona , Silva Anushka , Walsh Craig , Kessenbrock Kai 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells with potent immune suppressive activity. MDSCs regulate anti-tumor immunity by suppressing T cell proliferation. MDSCs can be further classified into granulocytic MDSCs (G-MDSCs) and monocytic MDSC (M-MDSCs). However, there is a lack of MDSC-specific markers since MDSCs, normal neutrophil granulocytes, and monocytes are defined by the same flow cytometry markers (CD11b+Gr1+). We used a breast cancer mouse model (MMTV-PyMT) to define cellular and molecular properties of MDSCs in single cell resolution. To test the capacity of MDSCs to inhibit immune responses, CD11b+Gr1+ cells from PyMT and wild type (WT) mice were sorted by fluorescence-activated cell sorting (FACS) from bone marrow, lung, and spleen, and then subjected to a T cell activation assay in co-culture with T cells. We found that predominantly spleen-derived CD11b+Gr1+ cells from PyMT significantly suppressed CD4 and CD8 T cell proliferation, while CD11b+Gr1+cells from bone marrow and lung showed no effect on T cell proliferation. Thus, we focused on spleen-derived CD11b+Gr1+ cells and utilized single cell RNA sequencing (scRNAseq) to compare breast cancer derived MDSCs to the respective cell populations harvested from normal, and non-tumor bearing hosts. Our computational analysis of 14,646 single cell transcriptomes revealed a separate G-MDSCs cluster from normal neutrophils and a M-MDSCs cluster from normal monocytes and a distinct gene expression signature for the entire MDSCs population. Our studies provide crucial insights into the biology of MDSCs, which may ultimately form the basis for novel marker and therapeutic avenues to improve cancer immunotherapy.
PLD3 and PLD4 are single-stranded acid exonucleases that regulate endosomal nucleicacid sensing
Authors: Nemazee David , Gavin Amanda L , Huang Deli 
Published: January 2023

Abstract
Genome-wide association studies have linked the gene encoding phospholipase D3 (PLD3) to Alzheimer’s disease and have linked PLD4 to rheumatoid arthritis and systemic sclerosis. PLD3 and PLD4 are endolysosomal proteins that we recently discovered are nucleases. PLD4-deficient mice have an inflammatory disease, marked by elevated levels of interferon-γ (IFN-γ) and splenomegaly. These phenotypes are blocked in the absence of TLR9. PLD3−/− mice have a relatively benign phenotype. Mice deficient in both PLD3 and PLD4 developed lethal liver inflammation in early life, which was blocked on a Unc93b-3d/3d background. These data suggest that PLD3 and PLD4 enzymes play important roles in the regulation of endosomal TLR signaling.
Mechanisms for secretion of antibodies with “anti-self” reactivity by the human obese adipose tissue
Authors: Frasca Daniela , Diaz Alain , Romero Maria , Blomberg Bonnie 
Published: January 2023

Abstract
We have shown that the human obese subcutaneous adipose tissue(SAT)contributes to systemic and B cell intrinsic inflammation and increased secretion of autoimmune antibodies. Immune cells are recruited to the SAT by chemokines released by both adipocytes and infiltrating immune cells. We have identified several mechanisms responsible for the release of “self” antigens in the human obese SAT, induction of class switch and production of autoimmune antibodies, such as hypoxia, NK cell cytotoxicity and DNA damage, which induce cell death and further release of pro-inflammatory cytokines. All these processes, in the context of an inflammatory environment like the obese SAT, stimulate the production of autoimmune IgG antibodies which have been described to be pathogenic. We have identified “self” antigens that stimulate the secretion of autoimmune antibodies in cultures of SAT-derived immune cells, using a Protein G immunoprecipitation protocol to purify the autoantibodies, combined with mass spectrometry. Results show that around 25% of these “self” antigens are found at high frequency in all the individuals evaluated so far in independent experiments. Moreover, &gt;90% of these “self” antigens (lipid and protein antigens, DNA, RNA) are cytoplasmic, nuclear or membrane-associated. These results allow the identification of novel pathways to manipulate in order to decrease pathogenic immune responses during aging.
The unfolded protein response contributes to antiproliferative and proapoptotic effects of IFN-λ4 in human hepatic cells
Authors: Onabajo Olusegun O , Wang Fang , Florez-Vargas Oscar , Obajemu Adeola , Vargas Joselin , Prokunina-Olsson Ludmila 
Published: January 2023

Abstract
IFN-λ4 is a type-III interferon with strong clinical significance in humans. Only individuals who carry the ΔG allele of a genetic variant rs368234815-TT/ΔG are genetically able to produce IFN-λ4 protein. Carriers of the ΔG allele have impaired ability to clear hepatitis C virus (HCV) infection, however the mechanisms are not fully understood. Here, we used RNA-seq to evaluate the effects of IFN-λ4 and IFN-λ3 expression in inducible hepatic cell lines (HepG2) with and without the receptor (IFNLR) required for signaling of both interferons. Pathway analysis showed that despite being poorly secreted, IFN-λ4 expression induced a stronger ISG response compared to IFN-λ3. IFN-λ4 expression was also associated with reduced proliferation and increased expression of genes involved in the unfolded protein response (UPR). Subcellular localization analysis revealed that IFN-λ4 was accumulated in the Golgi and lysosomes, suggesting it is misfolded and targeted for elimination. Furthermore, intrinsic but not extrinsic expression of IFN-λ4 inhibited cell proliferation via cell cycle arrest leading to apoptosis; both indicative of UPR outcomes. In conclusion, our results show that in addition to inducing ISGs, IFN-λ4 expression induces UPR, contributing to inhibition of cell proliferation. This could have important consequences for conditions such as HCV and fibrosis, where IFN-λ4 expression has been implicated.
Plasmacytoid dendritic cells contribute to pro-inflammatory and pro-fibrotic milieu in lung fibrosis
Authors: Singh Ram Raj , Valera Isela 
Published: January 2023

Abstract
Fibrosis, the end-result of tissue injury in a wide range of diseases, contributes to &gt;40% of mortality in the US. Plasmacytoid dendritic cells (pDC) are reduced in the peripheral blood, but increase in the lungs and skin of patients with systemic sclerosis, a prototypic fibrotic disease (JCI Insight 2018). The frequency of pDCs in the lungs of patients correlated with the severity of lung disease and with levels of proteins implicated in inflammation and fibrosis. Importantly, depletion of pDCs ameliorated lung and skin fibrosis in the bleomycin-induced animal model. pDC depletion also altered the expression levels of proteins and genes implicated in chemotaxis, inflammation, and fibrosis in the lungs. To test if pDCs directly contribute to inflammatory/profibrotic milieu in fibrosis, we injected C57Bl/6 mice with bleomycin, harvested lungs, and analyzed lungs for proteins by Western blot, ELISA, and flow cytometry. CD36 that is implicated in platelet-collagen adhesion, oxidative stress and inflammation was increased on pDCs but not on mDCs, other myeloid cells, B cells and T cells in the lungs of bleomycin-injected mice as compared to control mice. TLR7 was also increased more on pDCs than on all other immune cells examined, whereas TLR2 was increased more on mDCs and other myeloid cells but not on pDCs and B cells. TGFβ-latent peptide was higher on all immune cells examined in the lungs of bleomycin-injected animals than in controls. Among chemokine receptors, CCR2, CCR3, CCR6, CCR9 and CXCR4 were higher on pDCs than other cells. In summary, pDCs elicit profibrotic milieu in the development of systemic fibrosis. These observations identify the increased trafficking of pDCs’ to the affected organs as a therapeutic target in fibrosis.