bluuebunny/crossref_metadata_2025_split huggingface dataset:
A mechanistic base for alternative splicing of human immune system-derived thyroid stimulating hormone (TSH) β-subunit.
Authors: Klein John R. 
Published: January 2023

Abstract
Thyroid stimulating hormone (TSH) is a glycoprotein hormone consisting of an α and a β chain. Within the hypothalamus-pituitary-thyroid axis, TSH is produced by thyrotropic cells in the anterior pituitary. Upon release into the circulation, TSH binds to receptors in the thyroid, resulting in the release of triiodothyronine (T3) and thyroxine (T4). T3 and T4 regulate nearly every aspect of mammalian physiology, including basal metabolism, growth and development, mood, and cognition. Although TSHβ has been known for years to be produced by cells of the immune system, the functional significance of that has remained unclear. My laboratory identified a splice variant of human TSHβ (TSHβv) that consists of a truncated but biologically active portion of TSHβ made from exon 3 with the deletion of exon 2. The signal peptide of TSHβv is coded for by a portion of the 3′ end of intron 2. TSHβv is primarily produced by bone marrow cells and peripheral blood leukocytes, particularly though not exclusively by cells of the myeloid/monocyte lineage. An unresolved question pertains to the molecular mechanisms that account for TSHβv alternative splicing, Here, a model is presented whereby TSHβv is made through a series of unique genetic reorganization steps. These findings have implications not only for understanding how the TSHβv isoform is made, but also for gaining insights into the involvement of immune system TSHβ in health and disease.
MRP14 as an important regulator of macrophage function in type 2 diabetes mellitus
Authors: xia chang , Rao Xiaoquan , Wang Yunmei , Simon Daniel I. , Rajagopalan Sanjay , Zhong Jixin 
Published: January 2023

Abstract

Background
Myeloid-related protein14 (MRP14) plays an essential role in myeloid cell-mediated innate immune function and a number of inflammatory diseases. However, the role of MRP14 in obesity-induced inflammation and insulin resistance is not well defined. This study investigated the role of MRP14 in macrophage inflammation and obesity-induced insulin resistance.

Methods and Results
WT and MRP14−/− mice on C57BL/6 background were fed a high-fat diet or normal chow for 12 weeks. Tissue-resident macrophages in both adipose tissue and liver from obese mice expressed higher levels of MRP14. MRP14−/− mice showed a significantly improved insulin sensitivity, as measured by intraperitoneal glucose tolerance test and insulin tolerance test, compared to WT mice. There were reduced NLRP3 expression and caspase-1 activation in MRP14−/− macrophages after LPS stimulation compared to WT macrophages. In consistency, treatment of exogenous recombinant MRP8/MRP14 complex induced NFκB and caspase-1 activation. Glycolysis and mitochondrial respiration assay of macrophage were similar between WT and MRP14−/− mice. Activated MRP14−/− macrophages produced less T cell chemokines CCL5, CXCL9, and CXLL10, accompanied by reduced NFκB activation. Depletion of MRP14 from the supernatant decreased the activation of NFκB and production of chemokines (CCL5, CXCL9, and CXLL10) in WT macrophage but not MRP14−/− macrophage. Treatment of MRP8/MRP14 complex induced NFκB activation and increased the expression of CCL5, CXCL9, and CXLL10. In addition, T cells showed reduced migration towards the supernatant from MRP14−/− macrophages.

Conclusion
MRP8/14 regulates chemokine production from macrophage and T cell migration.
Statins regulates inflammatory macrophage phenotype through the activation of AhR
Authors: Manni Giorgia , Gargaro Marco , Turco Antonella , Scalisi Giulia , Matino Davide , Pirro Matteo , Fallarino Francesca 
Published: January 2023

Abstract
Statins are 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase competitive inhibitors, that lower blood cholesterol, but also exert potent anti-inflammatory effects. The xenobiotic receptor, aryl hydrocarbon receptor (AhR), plays crucial role in the control of inflammatory responses. Evidences suggest that selected statins promote the transcription of the main AhR target gene, cytochrome P450 family 1 (Cyp1a1). Macrophages are essential components of innate and adaptive immunity, characterized by AhR expression and activity. Based on these evidences, we investigated potential AhR-mediated immune regulatory properties of statins in Bone Marrow Derived Macrophages (BMDM).
We found that BMDM activation with the pro-inflammatory cytokine TNFa induced the inflammatory marker, inducible nitric oxide synthase (iNOS), which was completely prevented in BMDMs treated with TNFa in combination with specific statins, namely, atorvastatin and rosuvastatin. We confirmed high AhR expression in BMDM that was increased upon treatment with TNFa. We investigated the ability of different statins to activate AhR using a luciferase reporter assay cell lines and in BMDCs. We found that selected statins, such as atorvastatin and rosuvastatin, activated AhR in a dose dependent manner, an activation prevented in cells expressing selected AhR mutants. Notably, AhR deficient BMDMs, expressed undetectable levels of the main statin target protein HMG-CoA, which was instead highly expressed in wild type BMDMs.
These data demonstrate that natural or synthetic statins can play regulatory roles in macrophages. Moreover, our results suggest a potential involvement of AhR in statin-mediated anti-inflammatory effects in macrophages.
Distinct iron homeostasis in Th1- and Th2-biased mouse strains
Authors: Xiao Xia , Saha Piu , Yeoh Beng San , Kumar Matam Vijay 
Published: January 2023

Abstract
Iron is a proinflammatory transition metal ion, yet limited information is documented on the differences in iron homeostasis under Th1 and Th2 biased conditions. We used C57BL/6 (BL6) and Balb/C (BB) strains, which exhibit prototypical Th1 and Th2 immune responses, to investigate the extent to which Th1 and Th2 biased responses dictate iron homeostasis during healthy and inflammatory conditions. Systemic and tissue iron levels were assessed in age and gender matched BL6 and BB mice. Gene expression for iron absorption, transport, and storage proteins were quantified in duodenum and liver. Intracellular labile iron pool (LIP) was assessed in bone marrow-derived macrophages (BMDM) and neutrophils (BMDN). Hypoferremia of inflammation was assessed by measuring serum iron before and after LPS treatment. BB mice displayed significantly higher levels of systemic and storage iron than BL6 mice. Although the expression of hepcidin and ferroportin were comparable between the two strains, hepcidin-independent iron absorption is more prominent in BB mice, as indicated by higher expression of genes involved in iron absorption and transport: DcytB, DMT1, SFT, and hephaestin. Similarly, hepatic TfR1, TfR2, transferrin, and DMT1 expression was higher in BB mice. Moreover, the basal levels of LIP in BB BMDM and BMDN were 3.4 and 2.6 folds higher than BL6, respectively. The LPS-induced hypoferremia of inflammation is more pronounced in BL6 than BB mice. Taken together, iron homeostasis is distinct between these two strains during both healthy and inflammatory conditions. BB mice showed more iron load than BL6 mice, which could be explained, in part, by the higher expression of iron uptake genes and also reduced Th1 responses in BB mice.
Aggravated gut inflammation in mice lacking the taste signaling protein α-gustducin
Authors: Wang Hong , Feng Pu , Chai Jinghua , Redding Kevin , Margolskee Robert F. , Huang Liquan 
Published: January 2023

Abstract
Inflammatory bowel disease (IBD) is a debilitating immune-related condition that affects over 1.4 million Americans. Recent studies indicate that the taste system is involved in much more than sensing food flavor, and taste receptors have been localized in a variety of extra-oral tissues. One of the newly revealed functions of taste receptors and signaling proteins is modulation of immune responses to microbes and parasites. We previously found that components of the taste receptor signaling pathway are expressed in subsets of the intestinal epithelial cells. α-Gustducin, a key G-protein α subunit involved in sweet, umami, and bitter taste receptor signaling, is expressed in the intestinal mucosa. In this study, we investigated the role of α-gustducin in regulation of gut mucosal immunity and inflammation using gustducin knockout mice in the dextran sulfate sodium (DSS)-induced IBD model. DSS is a chemical colitogen that can cause intestinal epithelial damage and inflammation. We analyzed DSS-induced colitis in gustducin knockout versus wild-type mice after administration of 3% DSS in drinking water to mice. Our results show that the knockout mice had increased weight loss, diarrhea, intestinal bleeding, and inflammation over the experimental period compared to wild-type mice, concurrent with increased expression of TNF and IFN-γ but decreased expression of IL-13 and IL-5 in the colon. These results suggest that the taste receptor signaling pathway may play critical roles in regulating gut immune balance and inflammation.
In TH17-associated ileal inflammation, IFNγ and IL-17A support mucosal homeostasis and modulate self-tolerance
Authors: De Ciantis Christopher L. , Jeschke Jonathan C. , Mayne Christopher G. , Ziegelbauer Jennifer , Singh Selena , Suchi Mariko , Iwakura Yoichiro , Drobyski William R. , Salzman Nita H. , Williams Calvin B. 
Published: January 2023

Abstract
Crohn’s Disease is characterized by dysregulation of the immune response in the ileum. To study the perturbation of homeostasis within the ileum we utilize a “Bigenic” mouse model combining a TCR transgene and matching transgenic target antigen restricted to ileal crypt expression. Juvenile Bigenic mice experience TH17-associated ileitis and ileal hyperplasia. Antigen transcript is demonstrated to be localized exclusively in ileal crypts, as visualized with in situ hybridization of mRNA. Roughly 50% of Bigenic mice mature and acquire symptomatic phenotype (BigenicS) including colitis and suboptimal growth, while losing ileal hyperplasia. The remaining non-symptomatic Bigenic mice (BigenicNS) retain ileal hyperplasia and successfully build a supply of TH17 and ileal-reactive Treg cells.
Previous studies in our lab indicate that IL-17A and INFγ are protective in this Bigenic model – their depletion by antibody binding accelerates disease and hinders accumulation of ileal-reactive Tregs. Furthermore, INFγKO Bigenic mice demonstrate almost complete penetrance of the symptomatic phenotype, with near 100% fatality by 50 days of life. Manipulation of microbiota in juvenile INFγKO Bigenic mice can protect them from this dysregulated state. Administration of a streptomycin/bacitracin antibiotic cocktail ad libitum in drinking water upon initial manifestation of weight loss symptoms or initiated preemptively upon weaning rescues and prevents, respectively, the weight loss phenotype and colitis.
Identification of functionally unique CD4 T cells that are the circulating counterparts of epidermal resident memory T cells
Authors: Morawski Peter A. , Duhen Thomas , Klicznik Maria M. , Hoellbacher Barbara , Motley Samantha , Campbell Daniel J. , Gratz Iris K. 
Published: January 2023

Abstract
As a barrier organ, the skin contains specialized T cell populations that combat infection and also help maintain tissue homeostasis and promote wound repair. Circulating skin-tropic T cells can also be identified in the blood based on their expression of the cutaneous lymphocyte antigen (CLA), but the developmental and functional relationships between these circulating CLA+ T cells and tissue-resident T cells in the skin are not fully understood. Using 33-parameter Cytometry by time-of-flight (CyTOF) analysis we identified a novel population of skin-tropic CD4+ T cells in human blood that expresses the CD103 integrin and phenotypically and functionally resembles epidermal resident memory T cells (TRM) in the skin. RNA sequencing identified a set of signature genes shared by circulating CD103+CLAhi T cells in the blood and epidermal TRM cells in the skin, and suggested that CD103+CLAhiT cells contribute to normal skin function and response to tissue damage. Finally, using humanized NSG mice carrying full-thickness human skin grafts we demonstrated that, despite their identification as TRM, CD103+CLAhi T cells in the skin are capable of exiting the tissue and re-entering circulation. Thus, CD103+CLAhi T cells in the blood represent a circulating T cell population of cutaneous TRM, and this provides novel opportunities for the study and therapeutic manipulation of TRM cells in the contexts of cutaneous infection, inflammation, and tissue-repair.
Generation of IL-3 secreting T helper cells is associated with microbial challenge at barriers and IL-1 family of cytokines
Authors: Velayudhan Shajo Kunnath , Goldberg Michael F. , Saini Neeraj , Ng Tony , Johndrow Christopher , Johnson Alison Joy , Porcelli Steven A. 
Published: January 2023

Abstract
Naive CD4+ T cells differentiate into distinct T helper subsets upon T cell receptor stimulation in a unique cytokine environment. The cytokine environment leads to the expression of subset-specific master transcription factor which enables secretion of cytokines characteristic of that subset and suppression of cytokines associated with other T helper subsets. We recently described that cutaneous Mycobacterium bovis BCG vaccination in mice induces a subset of T helper cells characterized by the secretion of IL-3. Here, we report that the induction of this subset of T helper cells is route dependent; cutaneous but not intravenous immunization with M. bovis BCG induced these cells. Similar results were obtained with a model of cutaneous Herpes Simplex Virus II infection. Infection with Herpes Simplex Virus by vaginal route or with M. tuberculosis by aerosol route induced these cells suggesting mucosal sites are conducive to their generation. In these scenarios, IL-3 secreting cells co-expressed GM-CSF, IFNγ, TNF and IL-2. In vitro polarization experiments showed that IL-3 secretion by CD4+ T cells was facilitated by IL-1 family of cytokines and suppressed by cytokines that induce Th1 and Th2 helper cells. The characteristic cytokine expression pattern and the dependence on route and cytokine environment of these cells support the idea that IL-3 secreting CD4+ T cells may represent a unique T helper subset.
The immunological role of crypt olfactory neurons against neurotropic viruses in teleost fish
Authors: Sepahi Ali , Kraus Aurora , Galindo-Villegas Jorge , Kelly Cecilia , Garcia-Moreno Diana , Mulero Victoriano , Pau Mar Huertas , Salinas Irene 
Published: January 2023

Abstract
Vertebrate olfactory sensory neurons (OSNs) are located in direct contact with the external environment and therefore they directly exposed to microorganisms such as viruses. Teleost fish have four types of OSNs, including crypt neurons, a cell type with enigmatic function. Crypt neurons express tyrosine receptor kinase A (TrkA)-like molecule and ORA4. In this study, we hypothesized that TrkA expressing crypt neurons are fast sensors of viruses in the olfactory mucosa of fish serving as an ultra-rapid mechanism to elicit local antiviral immune responses. Our results show that, within minutes, nasal delivery of rhadboviruses induces caspase dependent apoptosis in crypt neurons along with neuronal activation in rainbow trout olfactory organ (OO) and olfactory bulb (OB). Induction of chemokine CCL19 and prostaglandin synthase expression was detected 15 minutes after nasal delivery of virus in OO; whereas, in parallel, significant downregulation of CCL19 expression was measured in the OB. Pharmacological blocking of TrkA abolished these responses. CD8+ T cells infiltrated the OO within minutes of nasal viral delivery and this response was abrogated when TrkA was blocked. We identified that the microvasculature surrounding the OB and not the peripheral blood is the source of the infiltrating CD8 T cells suggesting a rapid shunt of these immune cells from the OB microvasculature into the OO. In support, ablation of crypt neurons in zebrafish results in increased susceptibility to rhabdoviral challenge. Combined, our results indicate a novel function for a subset of teleost OSNs, crypt neurons, and the importance of sensory neurons as a first layer of danger detection in vertebrates.
The catalytic subunit of DNA-PK has a unique function in inflammation independently of Ku70 and DNA repair: a new opportunity to target the enzyme without interfering with DNA repair
Authors: Ghonim Mohamed Ahmed , Pyakurel Kusma , Luu Hanh , Okpechi Samuel , Ju Jihang , Boulares Hamid 
Published: January 2023

Abstract
Our laboratory demonstrated a critical role for DNA-dependent protein kinase (DNA-PK) in asthma pathogenesis via modulating the pertinent immune responses. DNA-PK is DNA repair enzyme composed of a catalytic subunit (DNA-PKcs) and two DNA-binding subunits (Ku70 and Ku80). Human cells express high levels of DNA-PK, surprisingly such high levels do not confer increased ability to repair DNA damage. Here we show that the role of DNA-PK in promoting inflammatory responses is independent of its function in the DNA repair. We examined the effect(s) of partial depletion of Ku70 on Ovalbumin (OVA) induced lung inflammation in mouse model of the disease. Of note, depletion of Ku70 by gene heterozygosity causes deficiency in DNA-PK-dependent DNA repair. Unlike the protective effects provided by DNA-PKcs gene heterozygosity, Ku70 heterozygosity did not alter the OVA-induced eosinophilia, mucus hypersecretion, Th2 cytokines production, or OVA-specific IgE upon OVA challenge. Interestingly, Ku70 heterozygosity enhanced methacholine-induced AHR over that of WT mice. Using a cell culture system, we demonstrate that while IL-4 and TNF-α are potent inducers of DNA-PKcs, such activation did not coincide with any detectable DNA damage or repair responses. Remarkably, while Ku70−/− blocked DNA-PKcs autophosphorylation in response to the DNA damage agent, etoposide, it did not affect the kinase in response to TNFα. Our findings suggest that the mechanism by which DNA-PK functions in inflammation is completely unrelated to its role in DNA repair, thus, unraveling a completely novel function for the kinase. More importantly, this provides a window of opportunity to target DNA-PK in inflammatory diseases without interfering with DNA repair processes.
Generation of specialized CD4 subsets: TFH and ThCTL, requires signals from local antigen & infection during the effector phase.
Authors: Devarajan Priyadharshini , Vong Allen M. , Castonguay Catherine H. , Bautista Bianca L. , Swain Susan L. 
Published: January 2023

Abstract
Certain key specialized tissue-restricted CD4 effector subsets such as cytotoxic T cells (ThCTL) in the infected tissue and follicular helper T cells (TFH) in secondary lymphoid organs reach their peak responses after other CD4 effectors (Th1, Th17). Using an influenza A virus (IAV) infection model, we find that generation of ThCTL and TFH require that CD4 effectors recognize specific Ag/MHC Class II, 5–7 days after initial infection, a phase we have termed the late Ag checkpoint. We used TcR transgenic CD4 donor T cells and host mice in which Ag presentation is restricted to specific APC, as well as addition of various APC subsets at the checkpoint, to study which APC drive TFH vs. ThCTL generation. While tissue-restricted antigen presentation is crucial, several APC subsets were able to present Ag effectively to drive TFH and ThCTL generation. However, we found that host infection during the late Ag checkpoint was needed for optimal generation of donor ThCTL and TFH subsets. This contrasts with most CD4 memory generation that is not dependent on infection. Thus multiple CD4 fate decisions are made at the late Ag checkpoint. Both optimal specialized effector and most memory generation require continuing Ag presentation, while further tissue-specific effector differentiation requires local Ag presentation and we think, signals from pathogen recognition pathways (PRR). We suggest that to induce optimum protective immune responses, vaccines and immunotherapies may need to be designed taking into account these requirements for additional Ag presentation and PRR pathways at the late Ag checkpoint.
Immortalization of splenic and peripheral blood macrophages using a multi-cistronic v-Raf/v-Myc lentivirus.
Authors: Beauregard Chelsi Ryan , Broder Christopher C. , Schaefer Brian C. 
Published: January 2023

Abstract
We have created an improved system for immortalizing mature macrophages by modifying the original v-raf/v-myc (J2) virus. Previously, these J2 viruses required a producer co-culture condition and maintained replicating virus. This provided more complexity, reagents, and biosafety risks. Our improved virus is replication deficient. Its lentiviral genome contains the v-raf/v-myc oncogenes and a thy1.1 surface marker, each of which is separated by a self-cleaving peptide. This virus, therefore, eliminates the issues described above. We transduced our modified virus into total murine PBMC isolated from murine peripheral blood or spleen. We found the transduction and successful cell line production can be done with as few as 1×105 PBMC. The resulting cytokine independent macrophage cell line from both spleen and blood expressed mature macrophage markers (F4-80+CD11b+) but lacked early myeloid progenitor markers (Sca1−c-kit−). In addition, we found these macrophages were functional as demonstrated by their ability to undergo phagocytosis and ability to upregulate MHCII in response to IFNγ. These findings demonstrate that the v-raf and v-myc oncogenes in our new system can immortalize macrophages from spleen but uniquely, and perhaps more importantly, can immortalize macrophages from blood. This advance provides a non-invasive method to obtain the starting tissue needed to make these macrophage cell lines. Having the v-raf/v-myc oncogenes in a lentivirus means it is likely this method will work on blood from humans and other species. Additionally, the cytokine independence associated with cell lines made with our virus make it ideal for creating macrophage cell lines from mammals without commercially available growth factors.
Using machine learning and RNA-seq to increase the accuracy and decrease the invasiveness of diagnosing eosinophilic esophagitis.
Authors: Lin Edwin , Flygare Steven , Peterson Kathryn , Clayton Frederic , Yandell Mark 
Published: January 2023

Abstract
Eosinophilic esophagitis (EoE) is an esophageal disease characterized by dysphagia and food impaction triggered by food or drug hypersensitivity with a prevalence of ~1/1,000 people. A conclusive diagnosis of EoE requires endoscopy and multiple biopsies, followed by eosinophil counts in multiple high-power fields. This procedure is painful and expensive, and due to the focal nature of the lesions, misdiagnoses are possible if biopsies do not sample enough diseased tissue. RNA sequencing allows the entire transcriptome within a biological sample to be assayed, but due to the vastness of the data, bioinformatics tools are required for meaningful interpretation. We have trained different machine learning classification algorithms and characterized their performance. We show that accurate classification is generalizable to biopsies that were not controlled for depth and were sequenced using different protocols and machines. Furthermore, we assess whether EoE status can be predicted using buccal epithelial tissue biopsies, which can be collected non-invasively but lack histological change. Together, these results show that machine-learning algorithms based on RNA-seq data could enable more accurate diagnosis of EoE using less invasive and lower-cost biopsy protocols.