bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantification of hTERT alternatively spliced isoforms in resting and activated human T cell subsets
Authors: Patrick Michael S. , Cheng Nai-Lin , An Jie , Weng Nan-ping 
Published: January 2023

Abstract
The activity of telomerase is regulated at multiple levels of the reverse transcriptase (hTERT) subunit. In T cells, it is well documented that telomerase activity is not detected in resting cells while it is high in activated cells. However, it is unclear whether full-length (FL) hTERT mRNA and AS products (ASPs) are expressed in resting T cells and how they affect T cell differentiation and function. Here, we analyzed hTERT mRNA (full-length and 3 types of ASPs) in CD4 and CD8 T cell subsets (naïve, central and effector memory) isolated from 66 healthy human adults. We detected hTERT FL mRNA in resting CD4 and CD8 T cell subsets and found that the level of hTERT FL mRNA decreased from naïve (N) to central memory (CM) to effector memory (EM) cells. Furthermore, hTERT FL mRNA levels were higher in CD4 than in corresponding CD8 subsets and undetectable in CD8 EM of most donors. In vitro stimulation with anti-CD3/CD28 antibody-coated beads induced a high level of hTERT mRNA in all subsets without changing the order of hTERT abundance in CD4 and CD8 T cell subsets. Finally, we measured telomerase activity in these T cell subsets and confirmed that resting T cells did not have detectable telomerase activity, but activated T cells did and the levels of telomerase activity concurred with expression levels of hTERT FL mRNA. In conclusion, quantifying hTERT FL and ASPs in T cell subsets confirmed no telomerase activity in the presence of FL hTERT mRNA in resting T cells and revealed a decrease of hTERT levels with T cell differentiation from naïve to memory. Currently, we are assessing which transcription factors are responsible for regulating hTERT expression levels in T cell differentiation.
LPS-elicited TSLPR expression enriches a functionally discrete subset of human CD14+ CD1c+ monocytes
Authors: Borriello Francesco , Iannone Raffaella , Di Somma Sarah , Vastolo Viviana , Petrosino Giuseppe , Visconte Feliciano , Raia Maddalena , Scalia Giulia , Loffredo Stefania , Varricchi Gilda , Galdiero Maria Rosaria , Granata Francescopaolo , Del Vecchio Luigi , Portella Giuseppe , Marone Gianni 
Published: January 2023

Abstract
Thymic stromal lymphopoietin (TSLP) is a cytokine produced mainly by epithelial cells in response to inflammatory or microbial stimuli and binds to the TSLP receptor complex, a heterodimer composed of TSLP receptor (TSLPR) and IL-7 receptor α (CD127). TSLP activates multiple immune cell subsets expressing the TSLP receptor complex and plays a role in several models of disease. Although human monocytes express TSLPR and CD127 mRNAs in response to the TLR4 agonist LPS, their responsiveness to TSLP is ill-defined. We demonstrate that TSLP enhances human CD14+ monocyte CCL17 production in response to LPS and IL-4. Surprisingly, only a subset of CD14+ CD16− monocytes (TSLPR+ mono) expresses TSLP receptor complex upon LPS stimulation in an NF-κB- and p38-dependent manner. Phenotypic, functional and transcriptomic analysis revealed specific features of TSLPR+ mono, including higher CCL17 and IL-10 production and increased expression of genes with important immune functions (i.e. GAS6, ALOX15B, FCGR2B, LAIR1). Strikingly, TSLPR+ mono express higher levels of the dendritic cell marker CD1c. This evidence led us to identify a subset of peripheral blood CD14+ CD1c+ cells that expresses the highest levels of TSLPR upon LPS stimulation. The translational relevance of these findings is highlighted by the higher expression of TSLPR and CD127 mRNAs in monocytes isolated from patients with Gram-negative sepsis compared to healthy controls. Our results emphasize a phenotypic and functional heterogeneity in an apparently homogeneous population of human CD14+ CD16− monocytes and prompt further ontogenetic and functional analysis of CD14+ CD1c+ and LPS-activated CD14+ CD1c+ TSLPR+ monocytes.
Binding of small molecule to ASC leads to the suppression of NLRP3 inflammasome and acute gout symptoms
Authors: Lee Hye Eun , Yang Gabsik , Lee Joo Young 
Published: January 2023

Abstract
The NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome, which is composed of NLRP3, the adaptor protein ASC, and caspase-1, is closely linked to the pathogenesis of metabolic diseases. We investigated whether caffeic acid phenethyl ester (CAPE), an anti-inflammatory phytochemical, could modulate the activation of NLRP3 inflammasome and have beneficial effects on the related disease such as acute gout. In primary mouse macrophages, CAPE blocked NLRP3 inflammasome activation induced by monosodium uric acid (MSU) crystals. In a mouse air pouch inflammation model and an acute gout model, oral administration of CAPE suppressed MSU-induced caspase-1 activation and interleukin (IL)-1b production, correlating with the attenuation of inflammatory symptoms. CAPE inhibited inflammasome activation induced by ASC, but not NLRP3 nor caspase-1. CAPE directly associated with ASC, resulting in the blockade of NLRP3-ASC interaction. These results demonstrate a novel inhibitory mechanism by which small molecules regulate NLRP3 inflammasome targeting ASC, suggesting the therapeutic strategy for NLRP3-related inflammatory diseases.
Composition of the myddosome during the innate immune response
Authors: Gillen Joseph G , Nita-Lazar Aleksandra 
Published: January 2023

Abstract
Critical for the innate immune response, the myddosome is a complex of proteins characterized by the presence of myeloid differentiation factor 88 (Myd88). The myddosome acts to transfer signals from the toll-like receptor (TLR) proteins to TNF receptor-associated factor 6 (TRAF6). In addition to Myd88, the myddosome contains multiple copies of the interleukin-1 receptor associated kinases (IRAK) 2 and 4 and is theorized to form following stimulation of the TLR proteins. Using affinity purification – mass spectrometry analysis (AP-MS), we identified the Myd88-associated proteins in mouse immortalized bone marrow-derived macrophages before and after lipopolysaccharide (LPS) treatment. Prior to LPS treatment, we found the stable association of Myd88 to IRAKs 2 and 4 along with the inhibitory factors IRAK3 and Sterile alpha and TIR motif-containing protein 1 (Sarm1). Following LPS treatment, the inhibitory factors were lost and Myd88 associated to the downstream effector proteins, including TRAF6. In contrast to the model, our results suggest that the cellular myddosome is preassembled but only activates upon stimulation. Because of this, we are examining the mechanism(s) that regulate the activity of the myddosome. We are currently using AP-MS followed by phosphopeptide enrichment with MS/MS analysis to identify possible phosphorylation sites. Using these data, we will evaluate the role of phosphorylation on myddosome activity. In addition, we will use SILAC with AP-MS to examine myddosome assembly and protein phosphorylation during a time course. This research was supported by the Intramural Research Program of the NIH, National Institute of Allergy and Infectious Diseases.
Caspase-11-mediated HMGB1 release from the liver drives immune cell pyroptosis in sepsis.
Authors: Deng Meihong , Scott Melanie , loughran Patricia , Wang Qingde , Lu Ben , Billiar Timothy R 
Published: January 2023

Abstract
High mobility group box 1 (HMGB1) was identified as a damage associated molecular pattern in sepsis. Neutralization of the HMGB1 even 24 hrs after the onset of sepsis is protective in experimental models. However, the sources of extracellular HMGB1 in sepsis and its function are not known. To address these questions, we generated myeloid (Lyz-creX HMGB1loxp/loxp) and hepatocyte (HC)-specific (Alb-creX HMGB1loxp/loxp) mouse strains and subjected these mice to cecal ligation and puncture (CLP) sepsis. Whereas CLP led to significant and sustained increases in HMGB1 levels in wild type and myeloid-specific HMGB1−/− mice, the levels were 75% lower in HC-specific HMGB1−/− mice at 18 hrs. This indicates that the liver is dominant source of HMGB1 during sepsis. The HC-specific knockout mice also showed significantly higher survival rates, as well as decreased immune cell pyroptosis in the peritoneum, spleen, and gut. Interestingly, HMGB1 level increased in HC culture media after LPS stimulation, which was not associated with cell death. Furthermore, using Liquid chromatography–mass spectrometry, we found that the HMGB1 released from HC are mainly acetyl form, suggesting that HC actively releases HMGB1 after LPS stimulation. We also found that the release of HMGB1 from cultured LPS-treated HC was TLR4 and caspase-11-dependent. Co-culture of wild type but not HMGB1−/− HC with bone marrow-derived macrophages led to increased pyroptosis of the cultured macrophages in response to LPS in a caspase-11-dependent manner. Our findings indicate that HC release HMGB1 during sepsis in a process that caspase-11-dependent and that liver-derived HMGB1 drives immune cell pyroptosis in severe sepsis.
NOD2 deficiency exacerbates imiquimod-induced, psoriasis-like dermatitis
Authors: Singh Tej Pratap , Cipolla Ellyse , Singh Satya P , Farber Joshua M 
Published: January 2023

Abstract
Nucleotide-binding oligomerization domain containing protein 2 (NOD2) is a CARD domain-containing pattern recognition protein in which inactivating mutations can cause Crohn’s disease. Mechanisms that have been suggested to explain the inflammation in Crohn’s disease associated with loss of NOD2 function include inadequate intestinal antibacterial defense or, on the other hand, diminished inhibition of toll-like receptor (TLR) signaling. Psoriasis is a common autoimmune skin disease for which recent data show significant up-regulation of NOD2 in keratinocytes of the lesional skin. We investigated a role for NOD2 in a TLR-driven mouse model of psoriasis-like dermatitis in which IL-17A and IL-22 are critical contributors. Following topical application of the TLR7/8 agonist, imiquimod, as compared with wild-type controls, Nod2−/− mice showed increases in epidermal thickness, epidermal neutrophils, and numbers of α/β and γ/δ T cells capable of producing IL-17A and IL-22 as well as expression of mRNAs for IL-17 and IL-22 in the treated skin. In addition, we found increased levels of mRNAs for IL-1β, and for several IL-1-inducible proteins, including COX2 and the neutrophil chemoattractants CCL3, CXCL2, and CXCL3, and decreased levels of mRNA for IL-1RA in the imiquimod-treated skin of the Nod2−/− mice. Taken together, these data suggest that NOD2 may be limiting skin inflammation in psoriasis by regulating IL-1/TLR signaling, supporting an important immunomodulatory role for NOD2 in inflammatory disease.
Long noncoding RNA (lncRNA) Negative Regulator of Interferon Response (NRIR) is induced along with other Interferon Induced mRNA in primary NK cells of uninfected and HCV infected persons.
Authors: Zebrowski Elizabeth , Barriocanal Marina , Fortes Puri , Valadkhan Saba , Anthony Donald D 
Published: January 2023

Abstract

Background
Natural Killer cells are thought to participate in control of HCV during the early phase of infection (first 6 months) and during IFN containing HCV therapy. At the same time, IFN resistance underlies wide variability in therapeutic response to IFN containing HCV therapy. We have previously described the lncRNA NRIR to be induced in hepatocytes upon IFNa exposure, and expression of NRIR was found to be responsible for down-modulation of the IFNa response. Whether NRIR is expressed within primary NK cells or participates in the IFN response within these cells has not been known.

Methods
Here we evaluated freshly isolated and IFNa treated primary human NK cells from 8 uninfected donors and 8 chronic HCV infected donors for NRIR expression by qPCR.

Results
We found NRIR expression in 100% of freshly isolated NK cells of uninfected donors and 40% of HCV infected donors. Upon IFNa exposure NRIR was greatly induced in 100% of uninfected donor and 67% of HCV infected donor NK cells.

Conclusions
These data indicate NRIR mediated regulation of the IFN response may be operative in primary NK cells of humans with or without HCV infection. Further studies are needed to determine the effect of NRIR on IFN induced NK cell function.
TLR2 and Adenosine Receptor A2a Induce ERK1/2 and c-FOS in Oral Squamous Carcinoma Cells
Authors: Kurago Zoya B , Palani Chithra , Ramanathapuram Lalitha , Lam-ubol Aroonwan 
Published: January 2023

Abstract
Squamous cell carcinoma (SCC) is a tenacious cancer prevalent at mucosal and skin surfaces. The level of oral (O)SCC differentiation does not correlate with prognosis, while tumor colonization by microorganisms and chronic inflammation may well contribute to OSCC pathogenesis. Microbial pattern recognition toll-like receptors (TLR) in immune cells induce inflammatory mechanisms, as they activate NF-kB and MAPK ERK1/2 pathways. Activated immune cells downregulate adenosine receptors (AR) A1 and A3, but upregulate inhibitory AR A2a and A2b, which also engage ERK1/2 when stimulated by adenosine. Malignant squamous cells express functional TLR2 and TLR4 as well, but their roles in OSCC cells are poorly understood. To address this gap, we evaluated TLR and AR expression and function in OSCC cells and keratinocytes. Five out of six OSCC lines expressed more TLR2 than TLR1, 4 or 6. TLR2, but not TLR4 stimuli induced AR A2a upregulation, while A2b did not increase. AR A1 and A3 mRNA was not detected in any oral squamous cells. TLR2 and A2a were also expressed in SCC cells and in dysplastic epithelial cells of all 17 archival oral specimens evaluated (use of specimens was approved by AU IRB). TLR2/1, TLR2/6 and AR stimuli induced ERK1/2 phosphorylation and accumulation of c-FOS in TLR2-responsive OSCC, which was similar to the effect of epidermal growth factor (EGF) on these cells. TLR2 and AR stimuli in responsive OSCC also stimulated the expression of the proliferation marker Ki-67. We conclude that products specific for TLR2 and A2a, such as bacterial lipopeptides and adenosine/inosine present in the tumor microenvironment, may contribute to OSCC progression via direct effects on malignant cells.
Comparison of the immunomodulatory effects of 3,4-dichlorophenyl-propanamide (DCPA) and N-(3,4-dichlorophenyl)-N-methylpropanamide (N-MeDCPA)
Authors: Barnett John B , Bergdorf Kensey N , McCall Jamie L. 
Published: January 2023

Abstract
Glioblastoma multiforme (GBM) are stage IV brain tumors for which there is no curative treatment. GBM depend on Orai1 calcium channel signaling for migration, proliferation, and the stability of various proteins. Thus, blocking Orai1 channels may provide a target for chemo-ablation of the tumor. DCPA and N-MeDCPA block Orai1 channels; however, N-MeDCPA has less in vivo toxicity. N-MeDCPA inhibits proliferation and migration in the U251MG glioblastoma cell line in a dose-dependent manner by 1.8% to 98.6% and 12% to 48%, respectively. The well-known role of the Orai1 channel in T cell activation may affect its usefulness as a drug. At high concentrations, DCPA inhibits T cell cytokine production; therefore, a direct comparison of the immunomodulatory effects of DCPA and N-MeDCPA on T cell activity and phenotype ex vivo are in progress. Future experiments will determine if N-MeDCPA can modulate the anti-tumor immune responses at concentrations that would directly inhibit the tumor. These results will be key in experiments using a mouse xenograft model of GBM to determine direct tumoricidal effects. If N-MeDCPA inhibits inflammatory and T-cell responses that promote tumor progression, in addition to its direct antitumor effects, it may have a future role in the treatment of GBM.
Bacterial amyloid curli acts as a carrier for DNA to elicit an autoimmune response via TLR2 and TLR9
Authors: Tursi Sarah , Lee Ernest , Lee Michael , Mederios Nicole , Wilson Paul , Gallucci Stefania , Wong Gerard C , Tukel Cagla 
Published: January 2023

Abstract
Bacterial biofilms are associated with numerous human infections. The predominant protein expressed in enteric biofilms is amyloid curli which forms immunogenic complexes with DNA. Infection with curli-expressing bacteria or systemic exposure to purified curli-DNA complexes triggers autoimmunity via the generation type I interferons (IFNs) and anti-double stranded (ds) DNA autoantibodies. DNA complexed with curli stimulates Toll-like receptor (TLR) 9 through a two-step mechanism. First, the cross beta-sheet structure of curli is bound by cell-surface TLR2, enabling internalization of the complex into endosomes. After internalization, the curli-DNA immune complex binds strongly to TLR9, inducing production of type I IFNs. Upon stimulation of macrophages with curli-DNA complexes, only 5% of TLR2−/− macrophages harbored intracellular curli, suggesting that TLR2 drives the internalization of curli-DNA complexes. Suppression of TLR2 internalization led to a significant decrease in Ifnβ expression. Structural analysis using small-angle X-ray scattering revealed that incorporation of DNA into curli fibrils results in the formation of ordered curli-DNA immune complexes resulting in optimal spacing to TLR9. Production of anti-dsDNA autoantibodies and type I IFNS in response to curli-DNA fibers was attenuated in the TLR2−/−, TLR9 mutant, and TLR2−/− - TLR9 mutant mice compared to wildtype mice, suggesting that TLR2 and TLR9 are critical for the autoimmune immune response. Overall, our results identify a pivotal series of events that leads to the severe pro-autoimmune effects of amyloid-expressing bacteria.
Sand fly salivary proteins-parasite-skin interaction
Authors: De Castro Waldione , Serafim Tiago Donatelli , Oliveira Luiz Fabiano , Kamhawi Shaden , Valenzuela Jesus 
Published: January 2023

Abstract
Leishmaniasis is a vector-borne disease transmitted by infected sand flies. During infection, sand fly salivary proteins are inoculated into the host dermis together with Leishmania parasites where proteins from the extracellular matrix are present. In this context, we hypothesize that sand fly salivary proteins interact with the parasite and with proteins from the extracellular matrix. Our preliminary findings show that proteins from sand fly salivary gland homogenate bind to the surface of promastigote forms of the parasite Leishmania infantum. Using recombinant sand fly salivary proteins we identified two major salivary proteins, LJL143 and LJL04, as the main salivary proteins binding Leishmania parasite surface. Furthermore, by a Surface Plasmon Resonance assay we tested the interaction between salivary recombinant proteins with different extracellular proteins. From all the recombinant proteins tested, only LJL04 interacts with Fibrinogen, Fibronectin, Collagen I, Multimeric VTNC and Laminin. Our plans are to confirm the salivary protein-parasite-extracellular matrix biding in vivo, firstly by exposing Balb/c mice ears to uninfected L. longipalpis bites and by detecting the presence of salivary proteins in the ear of mouse dermis post-sand fly bites. As future direction we are also planning to use L. infantum (transgenic fluorescent parasite)-infected sand flies bites to verify the possible co-localization of parasite and sand fly salivary proteins in the skin. Here, for the first time we show interaction between sand fly salivary proteins and Leishmania parasites and sand fly salivary proteins and extracellular matrix proteins.
Anti-alpha-toxin immunoprohylaxis reduces disease severity against a Staphylococcus aureus full-thickness skin wound infection in immunocompetent and diabetic mice
Authors: Ortines Roger Vargas , Cheng Lily , Cohen Taylor S , Gami Abhishek , Dillen Carly A , Ashbaugh Alyssa G , Miller Robert J , Wang Yu , Tkaczyk Christine , Sellman Bret R , Miller Lloyd S 
Published: January 2023

Abstract
Staphylococcus aureus infection is a major complication of acute and chronic skin wounds that delays wound closure and healing. These infections are particularly problematic in diabetic foot ulcers as the infection can become invasive, leading to osteomyelitis that often necessitates limb amputation. Prior reports in S. aureus skin infection models have indicated that neutralizing S. aureus alpha-toxin decreases disease severity by reducing skin lesion size and restoring an appropriate host immune response, however whether alpha-toxin neutralization impacts S. aureus-infected skin wounds is not clear, especially in the setting of diabetes. Herein, we evaluated the efficacy of MEDI4893* a neutralizing human anti-alpha-toxin monoclonal antibody (mAb) in a S. aureus skin wound infection model. Three full-thickness cuts were made on the back skin of wildtype (wt) C57BL/6 and diabetic (TallyHo) mice and the cuts inoculated with a bioluminescent methicillin-resistant S. aureus (MRSA) strain. In both WT and diabetic mice, prophylaxis with MEDI4893* resulted in a significant reduction in skin lesion sizes, decreased bacterial burden, improved re-epithelialization, and a decreased percentage of neutrophils and an increased percentage of macrophages in the skin compared with control mAb treatment. Thus, in S. aureus-infected skin wounds, anti-alpha-toxin mAb treatment reduced disease severity by not only inhibiting dermonecrosis but also by resolving neutrophilic inflammation.
Dectin-2 as a Primary Receptor for NLRP3-inflammasome Activation in Dendritic Cell Responses to Histoplasma capsulatum
Authors: Wu-Hsieh Betty , Chang Tzu-Hsuan , Huang Juin-Hua 
Published: January 2023

Abstract
Inflammasome is an intracellular protein complex that serves as cytosolic pattern recognition receptor (PRR) to engage with pathogens and to process cytokines of the interleukin-1 (IL-1) family into bioactive molecules. It has been established that interleukin-1β (IL-1β) is important to host defense against Histoplasma capsulatum infection. However, the detailed mechanism of how H. capsulatum induces inflammasome activation leading to IL-1β production has not been studied. Here, we showed in dendritic cells (DCs) that H. capsulatum triggers caspase-1 activation and IL-1β production through NLRP3 inflammasome. By reciprocal blocking of Dectin-1 or Dectin-2 in receptor-deficient DCs, we discovered that while Dectin-2 operates as a primary receptor, Dectin-1 serves as a secondary one for NLRP3 inflammasome. Both receptors trigger Syk-JNK signal pathway to activate both signal 1 (the transcription of Il1b) and signal 2 (activation of caspase-1). While both K+ efflux and cathepsin B (but not ROS) function as signal 2, viable H. capsulatum triggers profound lysosomal rupture leading to cathepsin B release. Our study demonstrates the role of Dectin-2 and Dectin-1 in host defense against H. capsulatum infection through induction of NLRP3 inflammasome activation and IL-1b production in DCs.