bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of Interleukin-6 in age-related frailty syndrome
Authors: Jergovic Mladen , Thompson-Ryder Heather L , Asghar Arveen , Nikolich-Zugich Janko 
Published: January 2023

Abstract
Aging is accompanied by chronic low-grade inflammation – a term which relates to increase in serum levels of cytokines such as interleukin 6 (IL-6), TNF-α and acute phase proteins such as C-reactive protein. Multiple studies in humans have shown that IL-6 most reliably increases with aging. Frailty is a geriatric syndrome characterized by decrease in physical ability and increased vulnerability to stressors. One of the most consistent findings in frail subjects is an elevation of serum IL-6 but we lack research which delineates is this a causational relationship. We developed a mouse model with inducible IL-6 expression (IL-6TET-ON/+ mice) that enables overexpression of IL-6 following stimulation with doxycycline (dox) administrated in food. IL-6 induction was dose dependent and led to increase in frailty as measured by 30-point murine clinical frailty index. Mice induced to express high levels of IL-6 quickly displayed an increase in frailty index, decrease in muscle grip strength and loss of fat. Mice induced to express 3–4 fold increase in IL-6 similar to frail humans displayed similar changes after months of induction. We measured IL-6 levels in serum and various tissues (gut, muscle, adipose, spleen) of aged (28-month-old) frail mice and adult controls as well as dox fed IL-6TET-ON/+ mice. We observed that IL-6 levels were increased in serums and spleen homogenates of aged mice and IL-6TET-ON/+ mice but not in other tissues. We determined that neutrophils were the main producers of IL-6 and their numbers were higher in spleens of aged and IL-6TET-ON/+ mice compared to adult controls. We conclude that elevated IL-6 serum levels are directly associated with age-related frailty and that spleen neutrophils are likely the main producers of IL-6.
JAK2V617F expressing macrophages stimulate IL-6 production through PKM1 mediated STAT3 activation
Authors: Ikezoe Takayuki , Li Rongqing , Sun Na , Chen Xin , Yang Jing 
Published: January 2023

Abstract
The JAK2V617F mutation is intimately involved in the pathogenesis of myeloproliferative neoplasms (MPNs) including polycythemia vera and essential thrombocytosis. The thrombotic events are serious complications occurring in individuals with MPNs which affect their prognosis. Interestingly, JAK2V617F is detected in some individuals who developed thrombosis without MPNs and thrombotic risk factors. These observations prompted us to hypothesize that JAK2V617F arising in myeloid cells could contribute to cardiovascular disease by promoting the production of proinflammatory cytokines. To test this, we established JAK2V617F expressing murine macrophages (JAK2V617F macrophages) and explored their function. We found that the levels of p-STAT3, one of downstream targets of JAK2, were markedly elevated in JAK2V617F macrophages in associated with an increase in the production of IL-6 at baseline and following stimulation with LPS, as measured by Western blotting and ELISA, respectively. While, inhibition of STAT3 by C188-9 significantly decreased the production of IL-6 in the culture medium harvested from JAK2V617F macrophages. In further experiments, JAK2V617F endowed macrophages with elevated glycolytic phenotype in parallel with aberrant expression of PKM1. Interestingly, silencing of PKM1 inactivated STAT3. On contrary, ectopic expression of PKM1 activated STAT3. Importantly, JAK2V617F contributed to PKM1 protein stabilization via blockade of lysosomal-dependent degradation via chaperone-mediated autophagy (CMA). All of these observations indicated that JAK2V617F could protect PKM1 from CMA-mediated degradation, leading to activation of STAT3, which promoted IL-6 production.
Nrf2 differentially regulates pro- and anti-inflammatory function of Th17 cells through RORγT, SOD3 and Ahr
Authors: Lin Xun , Tawch Suzanne , Gaudino Stephen , Roy Suyasha , Castillo Patricia , Elsegeiny Waleed Abdel Wahab , Wakabayashi Nobunao , Oury Tim , Pociask Derek , Chen Kong , Syritsyna Olga , Coyle Patricia , McLinskey Nancy , Melville Patricia , Awasthi Amit , Kolls Jay K. , Kumar Pawan 
Published: January 2023

Abstract
IL-17A and IL-22 derived from CD4+ T cells are significant for both inflammatory and tissue protective responses. However, it remains unclear whether IL-17A and IL-22 can be regulated differently to achieve targeted functional outcome.
Here, we showed that Nrf2 selectively regulated IL-17A and IL-22 responses in CD4+ T cells. Nrf2−/− mice challenged with Ovalbumin (Ova) + LPS displayed reduced IL-22 but enhanced IL-17A response. Interestingly, CDDO-Im, a selective Nrf2 activator, induced IL-22 but suppressed IL-17A response in CD4+ T cells polarized under Th17 cell condition. CDDO-Im-activated CD4+ T cells had lower Rorc, but increased Cybb, Sod1 and Sod3 transcript. Luciferase reporter assay showed that IL-17A promoter activity is inhibited by Nrf2 at the presence of RORγT, suggesting that inhibition of IL-17A by Nrf2 is RORγT-dependent. CDDO-Im-impaired IL-17A response was also abolished in SOD3−/−, showing that SOD3 is involved in CDDO-Im-mediated inhibition of IL-17A. Furthermore, we found that CDDO-Im induced aryl hydrocarbon receptor (Ahr) and its downstream Cyp1a1 and Cyp1b1 gene expression. Additionally, CDDO-Im induced Nqo1 expression, an Nrf2-activated gene, in WT mice but not in Ahr−/− mice. CDDO-Im-mediated induction of IL-22 production in CD4+ T cells was abrogated in Ahr−/− mice, supporting that Ahr is involved in the induction of IL-22 by Nrf2. Finally, upon the stimulation of anti-CD3 and anti-CD28, CDDO-Im also mediated the inhibition of IL-17A but induction of IL-22 production in PBMCs from relapsing-remitting multiple sclerosis patients.
Collectively, our data show that Nrf2 promotes IL-22 production via Ahr, whereas RORγT and SOD3 are involved in Nrf2-mediated inhibition of IL-17A expression in Th17 cells.
The role of IL-29 in obesity-induced inflammation and insulin resistance
Authors: HSU YU HSIANG , CHIU CHIAO JUNO , LIN TIAN YU 
Published: January 2023

Abstract
Adipocyte-macrophage crosstalk plays a critical role in regulating adipose tissue microenvironment and causes chronic inflammation in the pathogenesis of obesity. Interleukin-29 (IL-29), a member of type 3 interferon family, plays a role in host defenses against microbes. The macrophage is one of the cellular sources of IL-29; however, the role of IL-29 in metabolic disorders is still unclear. We explored the function of IL-29 in the pathogenesis of obesity-induced inflammation and insulin resistance. We found that serum IL-29 level was significantly higher in obese patients. IL-29 upregulated IL-1β, IL-8, and monocyte chemoattractant protein-1 (MCP-1) expression and decreased glucose uptake and insulin sensitivity in human SGBS adipocytes through reducing glucose transporter 4 (GLUT4) and AKT signals. Inhibition of IL-29 could reduce inflammatory cytokine production in the macrophage-adipocyte co-culture system, which mimics an obese microenvironment. In vivo, IL-29 reduced insulin sensitivity and increased the number of peritoneal macrophages in high-fat diet (HFD)-induced obese mice. IL-29 increased M1/M2 macrophage ratio and enhanced MCP-1 expression in adipose tissues of HFD mice. Therefore, we have identified a critical role of IL-29 in obesity-induced inflammation and insulin resistance, and we conclude that IL-29 may be a novel candidate target for inhibiting obesity-induced inflammation and insulin resistance in patients with metabolic disorders.
A Novel Viral Signaling Complex Containing Host Chemokine Receptor CXCR4, Interleukin-10 Receptor, and Human Cytomegalovirus pUS27
Authors: Spencer Juliet , Tu Carolyn 
Published: January 2023

Abstract
CXCR4 is a chemokine receptor that plays an essential role regulating cell movement during development and immune responses. Human cytomegalovirus (HCMV) is a widespread herpesvirus causing little disease except in immune compromised hosts, especially transplant recipients. HCMV establishes latency in myeloid cells and maintains lifelong infection by manipulating chemokine signaling, especially the CXCR4 pathway. HCMV US27 encodes a viral chemokine receptor (pUS27) that constitutively upregulates host CXCR4 expression, while UL111A encodes cmvIL-10, a homolog of cellular interleukin-10 (IL-10) that binds the IL-10 receptor (IL-10R). Downstream effects of cmvIL-10 include activating Stat3 and enhancing CXCR4 signaling responses to chemokine CXCL12. We used the proximity ligation assay (PLA) to examine the cellular distribution of host CXCR4 and IL-10R, and the results show the two receptors colocalized to discrete clusters on the cell surface. Exposure to CXCL12 caused increased receptor clustering and triggered calcium mobilization. We found that pUS27 was also closely associated with both CXCR4 and IL-10R in the same PLA clusters in transfected HEK293 cells and in HCMV-infected fibroblasts. The addition of CXCL12 and cmvIL-10 resulted in significantly higher receptor clustering, calcium flux, and cell migration of transfected and virus-infected cells. These results indicate that CXCR4, IL-10R, and pUS27 all associate to form a novel virus-host signaling complex that enhances signaling and chemotaxis to CXCL12 during HCMV infection.
CCR4 and CCR7 make distinct contributions to the spatial distribution and negative selection of different thymocyte subsets
Authors: Li Yu , Lancaster Jessica N , Nam Seo Hee , Seo Hyemin , Selden Hilary J. , Ehrlich Lauren Ilyse Richie 
Published: January 2023

Abstract
Central tolerance mechanisms, including clonal deletion and Treg selection, are required to remove autoreactive T cells generated during thymocyte differentiation. To ensure T cell tolerance to diverse self-antigens, the majority of the peptidome is displayed in a sparse mosaic by medullary thymic epithelial cells and dendritic cells in the medulla. Thus, thymocytes must efficiently enter the medulla and scan multiple antigen presenting cells to ensure complete tolerance. We and others have previously reported that the chemokine receptors CCR4 and CCR7 are important regulators of medullary enrichment, rapid motility, and negative selection of post-positive selection thymocytes; however, it is not known whether CCR4 and CCR7 play distinct or redundant roles in central tolerance. Here, we report that early post-positive selection thymocytes undergo chemotaxis towards CCR4 ligands, while progressively mature cells migrate towards CCR7 ligands. Synchronized positive selection experiments reveal that CCR4 is upregulated within hours of receiving a positive selection signal, while CCR7 upregulation is delayed for about 24h. 2-photon microscopy reveals that CCR4 facilitates entry of early post-positive selection cells into the thymic medulla. These findings suggest that CCR4 and CCR7 promote distinct stages of negative selection. Consistent with this possibility, flow cytometric analysis reveals that CCR4 deficiency results in a selective decline in early-stage negative selection, while CCR7 deficiency impairs only late-stage negative selection. Altogether, our data suggest a new model in which CCR4 and CCR7 mediate medullary entry of distinct thymocyte subsets and contribute differentially to central tolerance.
Reduced level of CX3CR1 positive T-cells and monocytes in children with, newly diagnosed, Type 1 diabetes
Authors: Dadfar Elham , Ghaderi Amineh , Moshfegh Ali , Barcenilla Hugo , Casas Rosaura , Samuelsson Ulf 
Published: January 2023

Abstract

Background
The chemokine CX3CL1 is associated with many autoimmune diseases. Type 1 diabetes (T1D) is an autoimmune disease characterized by an insufficient insulin production due to T-cells mediated destruction of pancreatic beta cells. CX3CL1/CX3CR1 plays an important role in recruitment and extravasation of T-cells and monocytes to the inflammatory site.

Aim
To investigate the role of CX3CL1 and its receptor CX3CR1 in children with T1D in acute phase (day 0), 10 days and 3 months after treatment compared to healthy subjects (HS).

Methods
Blood samples from children (range 2–18 years) with new onset type 1 diabetes (day 0 (n=14), day 10 (n=9) and 3 month (n=14)) and 24 age matched HS was collected. The concentrations of CX3CL1 in plasma (T1D day 0, day 10 and HS) was measured by ELISA. Receptor expression of CX3CR1 on monocytes and T-cells was analyzed by flow cytometry.

Results
The study showed significant higher concentrations of CX3CL1 in serum from T1D day 0 compared to HS (p&lt;0.05) and T1D day 10 (p&lt;0.01). No significant difference was observed between T1D at day 10 and HS. Furthermore, the level of CX3CR1 positive T-cells was significantly lower in T-cells from T1D day 0 compared to HS (p&lt;0.003) and T1D 3 months (p&lt;0.007). In addition, the amount CX3CR1 positive monocytes were significant lower in T1D day 0 compared to HS (p&lt;0.003).

Conclusion
The low amount of CX3CR1 positive T-cells and monocytes in peripheral circulation in acute phase T1D, despite the high level of s-CX3CL1, indicate that the T-cells and monocytes might have transmigrated to the inflammatory site. This study indicates that CX3CL1/CX3CR1 axis could play a role in the pathogenesis of T1D.
CRISPR-based identification of factors influencing IL-22 and IL-17 expression in type III innate lymphoid cells
Authors: Brown Rachel Anne , Dangel Andrew W , Saini Ankita , Oltz Eugene M. 
Published: January 2023

Abstract
Cytokine regulation of immune responses is influenced by multiple factors, including the target cell type and its microenvironment. Type III innate lymphoid cells (ILC3s) rapidly produce cytokines, including interleukin (IL)-22 and IL-17, in response to pathogens or injury. Dysregulation of IL-22 or IL-17 contributes to numerous pathologies, including autoimmune diseases and malignancies. Thus, a deeper understanding of the factors that regulate productive expression of IL-17/22 by ILCs or by their type III helper T cell counterparts is warranted.
We have used a genome-wide CRISPR interference screen to elucidate genes that drive IL-22/17 expression by ILC3s. We engineered the murine ILC3-like line, MNK3, to inducibly express catalytically dead Cas9 (dCas9) linked to the transcriptional repressor KRAB. Following lentiviral transduction of a sgRNA library, cells were stimulated with IL-23 and IL-1b. Guides from cell populations sorted by levels of IL-17/22 expression were sequenced and analyzed for depletion (target may be necessary for cytokine expression, e.g., il17, jak3, and il23r) and enrichment (target’s suppression may promote cytokine expression). We are currently validating “hits” that are components of several biological pathways, including transcriptional regulation, chromatin modification, notch/wnt and cadherin signaling, and RNA splicing. Results will also be presented for a complementary CRISPR activation screen. These data will contribute new insights into the biological processes necessary for proper expression of type 3 cytokines by innate and adaptive lymphoid cells.
IL-17 promotes protective antibody during gut infection through IκB-ζ-dependent LN stromal cell activation
Authors: Wu Dongwen , Majumder Saikat , McGeachy Mandy J 
Published: January 2023

Abstract
Fibroblastic reticular cells (FRCs) provide critical regulation of T and B cell. Our recent study shown metabolic reprogramming through Interleukin 17 (IL-17) was critical for the FRCs expansion and autoantibody production. Although studies shown IL-17 is important for B-cell activation and induction of IgA responses, the mechanisms have been unclear. We further hypothesized IL-17 signaling in FRCs is critical to drive protective antibody responses, and prior FRCs activation through a distinct inflammatory episode could enhance this response. We employed Dextran Sulfate Sodium (DSS) induced-colitis followed by Citrobacter rodentium infection model. Mice pretreated with DSS shown increased FRCs expansion and germinal center (GC) formation and enhanced anti-Citrobacter antibody production, leading to a quicker Citrobacter clearance, less colon inflammation and better outcome. IL17a neutralization in DSS phase blocked both the expansion of FRCs and the enhanced B cell response. We further confirmed FRCs failed to expand during Citrobacter infection in FRC-restricted IL-17RA deletion mice. These mice had reduced antibody production in both serum and feces, along with higher Citrobacter burden. We previously shown IL-17 induction IκB-ζ was required for metabolic reprogramming in vitro. Now, we found FRC-intrinsic IκB-ζ expression was required for in vivo Cpt1a (a rate-limiting transporter for fatty acid oxidation) expression, GC formation and antibody responses. RNAseq data suggest GC survival factors such as IL-6 and BAFF were increased by IL-17 signaling in FRCs. In conclusion, IL-17 indirectly promotes GC formation and antibody responses by inducing IκB-ζ expression in activated LN stromal cells during gut infection.
WGCNA identification of CXCL13 and associated genes involved in the Tumor Immune Microenvironment (TIME) of lung adenocarcinoma
Authors: Young Corey D , Dammer Eric , Nunez Sha’Kayla , Griffen Tiara , Carey Kaylin , Dill Courtney Denise , Lillard James W 
Published: January 2023

Abstract
Lung cancer is the second most commonly diagnosed cancer, causing the most cancer-related deaths in the United States. Non-small cell lung cancer (NSCLC) accounts for over 75% of lung cancer cases of which lung adenocarcinoma (LUAD) is the most common. Chemokines and their receptors have been shown to be highly expressed in a number of cancers, presumably leading to abnormal signaling. Elevated levels of CXCL13, and its receptor, CXCR5, have been correlated with the overall features of mortality, but the exact mechanisms in which these factors act to promote NSCLC progression are not fully understood. Here, we studied the role of CXCL13, CXCR5, and associated genes expressed in NSCLC tumor immune microenvironment (TIME). The software package WGCNA was used to establish clusters of highly correlated genes related to clinical characteristics. A total of 23,329 transcripts were used to identify 37 modules (gene co-expression networks). The light-cyan module, consisting of 491 genes contained both CXCR5 and CXCL13. The light-cyan module is a hub gene heavy module with the top 300 transcripts exceeding our hub gene threshold (kMe &gt; 0.6). The light-cyan module transcripts where negatively correlated with age. The top biological processes associated with the light-cyan module include immune system processes, regulation of lymphocyte activation, regulation of immune response and cellular defense response. Collectively, the gene clusters identified along with their clinical correlates provide a novel insight into the complex biology of NSCLC and its TIME.
Defining IL22 regulatory logic in type 3 lymphoid cells
Authors: Saini Ankita , Bando Jennifer K. , Collins Patrick L. , Cella Marina , Colonna Marco , Oltz Eugene M. 
Published: January 2023

Abstract
The cytokine IL-22 is a major effector expressed by type 3 immune cells (T helper 17, Th17 and type 3 innate lymphoid cells, ILC3s), and has diverse roles in maintaining intestinal homeostasis, defense against pathogens, cell proliferation, and tissue regeneration. Perturbations in the homeostatic levels of IL-22 have been associated with various inflammatory disorders and tumors. However, significant gaps remain in our understanding about the regulatory mechanisms controlling IL-22 expression by type 3 cells in health and disease.
In this regard, we generated a stable clone of Mnk3, a mouse ILC3-like cell line, which harbored a doxycycline-inducible version of catalytically dead Cas9 nuclease (dCas9) fused with KRAB repressor domain. We performed a CRISPRi screen in this dCas9-KRAB Mnk3 line by infecting it with a lentiviral library of sgRNAs targeting IL22 regulatory space. Our CRISPRi screen identified a set of candidate enhancers important for IL22 expression in type 3 cells. Chromatin profiling data suggested a differential pattern of activities for these enhancer elements in immune cell subsets. We have now deleted several enhancers from the germline of mice and will present results describing the impact of these mutations on IL-22 expression in type 3 lymphoid cells from the innate and adaptive immune systems.
Collectively, this study defines the cis-regulatory circuits controlling IL22 in type 3 immune cells and thus will open new avenues for therapeutic intervention in IL-22 pathologies.
Single-cell analysis maps the dynamic landscape of mouse iNKT cell development and identifies Cbfb as a key regulator for early commitment
Authors: Wang Jie , Adrianto Indra , Liu Tingting , Wu Xiaojun , Sant’Angelo Derek B , kronenberg mitchell , Zhou Li , Mi Qing-Sheng 
Published: January 2023

Abstract
Invariant natural killer T (iNKT) cells are innate-like T cell that share the characteristics of T cell and NK cell. iNKT cell acquire an effector function and differentiate into iNKT1, iNKT2 and iNKT17 sub-lineages prior to the thymic export. Although remarkable advancements in iNKT cell development have been made, the underlying molecular programs that guide their differentiation and heterogeneity remain unclear. Here, we profiled the transcriptomes of over 17,000 iNKT cells and the chromatin accessibility states of over 39,000 iNKT cells from the mouse thymus across stage 0 to stage 3 using scRNA-seq and scATAC-seq. By integrating iNKT cells transcriptome and chromatin accessibility profiles, we found that both iNKT2 and iNKT17 lineage commitment occur at stage 0, while iNKT1 cells initiate their differentiation as early as stage 1. Furthermore, iNKT1 and iNKT2 cells exhibit extensive phenotypic and functional heterogeneity, while iNKT17 cells are relative identical. Reconstruction of iNKT developmental trajectories revealed two possible pathways stemming from iNKT progenitors. More importantly, we identified Cbfβ (core-binding factor subunit beta) and known Egr2 and Slamf6 exhibit similar expression pattern in iNKT precursors. Deletion of Cbfβ in the mice blocked iNKT cell development at stage 0 and stage 1, and Cbfβ-deficient iNKT cells were failed to be differentiated into effector iNKT subsets. Bone marrow transfer experiments further indicated that the defective iNKT cell development in Cbfβ KO mice was cell-intrinsic. Overall, our findings revealed the cellular heterogeneity of iNKT cells, mapped the potential developmental trajectories, and identified Cbfβ as a key regulator for early iNKT cell commitment.