bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adeno-associated viral vector-mediated anti-inflammasome gene transfer alleviates asthmatic airway inflammation in OVA-sensitized mice model
Authors: Kuo Ming-Ling 
Published: January 2023

Abstract
Bronchial asthma is a chronic inflammatory disorder of the airway characterized by airway hyperresponsiveness (AHR), pulmonary eosinophilia, elevated serum IgE, and dysregulated T helper (Th) 2 cytokines. It is becoming urgent to identify target molecules for the preventive and therapeutic purpose. ATP accumulation in bronchoalveolar lavage fluid (BALF) of asthmatic subjects indicates that extracellular ATP and the downstream responses are involved. ATP is an activator of NLRP3 inflammasomes and tightly regulated by CD39, a nucleoside triphosphate diphosphohydrolase encoded by entpd1. Upregulated BALF ATP levels and decreased CD39 RNA were accompanied with asthmatic cardinal features in ovalbumin (OVA)-sensitized mice. Recombinant adeno-associated virus (rAAV) has been developed to carry exogenous genes with safe and low toxic therapeutic potential. The different serotypes and tropisms of AAVs benefit tissue specific gene targeting. This study aims for determining whether lung-targeting rAAV carrying CD39 (AAV/CD39) is able to reduce extracellular ATP in BALF and inhibit the OVA-induced allergic inflammation in an asthmatic murine model. The mice were sensitized and challenged with OVA. rAAV vectors were administrated into the lungs of mice 3 days before the first OVA exposure. The data indicated that ATP levels in BALF were downregulated accompanied with rescued CD39 expression. The administration of AAV/CD39 was able to suppress AHR, pulmonary eosinophilia, and the production of IL-5 from OVA-activated mediastinal lymph nodes (MLNs) cells. The treatment of AAV/CD39 attenuated the asthmatic airway inflammation and AHR. Collectively, ATP could be a potential target for gene therapy on asthma.
The Role of Regulator of G-Protein-Signaling 4 (RGS4) in Control of Airway Hyper-responsiveness in Asthma
Authors: Madigan Laura , Gordon Elizabeth , Ganesan Sundar , Levine Stewart , Druey Kirk M 
Published: January 2023

Abstract
Asthma is a common respiratory disease characterized by airway hyper-responsiveness (AHR), inflammation, and airway obstruction. In most patients, this disease can be managed with steroids and bronchodilators. However, these treatments are not effective for the 15%-25% of patients with severe asthma who may develop fixed airway obstruction due to increased airway smooth muscle (ASM) mass.
ASM cells control airway patency through G-protein coupled receptors (GPCRs). Namely, GPCRs can activate the IP3 pathway to induce calcium release and airway contraction. Regulators of G-protein signaling (RGS) are a family of proteins that are known to negatively regulate the activity of GPCRs. In ASM cells, RGS4 is relatively abundant compared to other RGS4 proteins. In people with asthma, increased RGS4 expression is observed in the group with greatest impairment in lung function.
We hypothesize that aberrant RGS4 expression and/or function contributes to AHR through dysregulated GPCR signaling in ASM cells. We studied bacterial artificial chromosome (BAC) transgenic (Tg) mice, which overexpress both RGS4 and eGFP under the control of endogenous RGS4 promoter. In models of acute asthma, Tg mice exhibited reduced AHR as assessed by measurements of lung resistance in live, anesthetized mice and contraction of precision cut lung slices. These findings suggest that RGS4 is an important controller of bronchial tone in asthma through its effects on contraction signaling in ASM cells.
Transgenic expression of H-2Db class I molecules influences the development of brain atrophy during Picornavirus infection
Authors: Huseby April M , Atanga Pascal A , Gamez Jeffrey D , Cumba-Garcia Luz M , Macura Slobodan I , Johnson Aaron J 
Published: January 2023

Abstract
Brain atrophy is a common feature of numerous neurological diseases in which the role of neuroinflammation remains poorly defined. We have previously demonstrated the development of brain and spinal cord atrophy in the Theiler’s murine encephalomyelitis virus (TMEV) model of multiple sclerosis. However, cellular and molecular mechanisms of brain atrophy remain poorly understood. We therefore evaluated the contribution of major histocompatibility (MHC) class I molecules in atrophy development during TMEV infection. To accomplish this, we created a novel transgenic FVB/NJ mouse by introducing the H-2Db (Db) class I molecule. Expression of Db class I molecule confers resistance to persistent TMEV infection and demyelination in the normally susceptible FVB/NJ strain (H2-Dq class I haplotype). Next, we compared the development of brain atrophy, assessed by volumetric analysis of T2-weighted MRIs, in FVB/Db mice to wild-type FVB/NJ mice following viral infection. FVB/NJ mice did not significant brain develop atrophy over 4 months, whereas significant brain atrophy was observed in the FVB/Db mice. FVB/Db mice also developed a significant increase in the overall number of CNS infiltrating CD8 T cells and a strong CD8 T cell response towards an immunodominant Db TMEV epitope, VP2121-130. A number of CD8 T cells were also observed in close proximity to virus infected neurons. We therefore propose a hypothesis that class I restricted CD8 T cell responses promote the development of brain atrophy. This model provides a unique opportunity to analyze the contribution of immune cells to neuronal loss and brain atrophy in a system where persistent virus infection and demyelination are not confounding variables.
Investigation of immune cell maintenance and function in MBL-sufficient and MBL-deficient individuals
Authors: Dean Melinda M , Luimstra Jolien , Rooks Kelly , Flower Robert L 
Published: January 2023

Abstract

Background and Aims
Mannan-binding lectin (MBL) is a mediator of innate immunity, deficiency is associated with increased severity of infection. Pathogen recognition and complement activation functions are well characterised, however, mechanisms behind the reduced overall response to pathogens in of MBL-deficient (MBL-D, defined as &lt;0.5μg/mL) individuals remains poorly understood.

Methods
Absolute counts of NK cells, neutrophils, monocytes, myeloid DC, plasmacytoid DC, lymphocyte subsets (including Tregs), and the concentrations of 12 inflammatory cytokines were assessed on freshly collected peripheral blood (healthy volunteers) and following 6 and 24 hours in vitro for MBL-sufficient (MBL-S, n=10) and MBL-D (n=11) individuals (unpaired T-test, 95% CI). Mixed lymphocyte response (MLR) infection models (LPS, zymosan) as well as intracellular staining were used to study the maintenance and function of T and B lymphocytes.

Results
We observed few differences between MBL-S and MBL-D individuals in immune cell maintenance and function. Of note, we found a significantly higher proportion of CD4+CD25+Foxp3+ Tregs in MBL-D individuals compared to MBL-S individuals following 3 days autologous MLR in both unstimulated and zymosan-stimulated cultures.

Conclusions
This study supports the hypothesis that in association with immune challenge MBL deficiency can result in poorer patient outcomes. Our finding of increased Tregs in MBL-D individuals warrants further investigation in the clinical setting, as a potential mechanism underpinning increased disease susceptibility and severity of infection in MBL-D individuals.
Distinct features of human CD8 T cell TCR repertoire specific to influenza A virus matrix protein M1
Authors: Chen Guobing , Yang Xinbo , Ko Annette , Gao Mingming , Zhang Yongqing , Shi Alvin , Sun Xiaoping , Mariuzza Roy A. , Weng Nan-Ping 
Published: January 2023

Abstract
CD8 T cells play a vital role in the immune response to viral infection. Diversity of the T cell antigen receptor (TCR) is believed to be essential for the host to survive in this microorganism-rich world. However, there is limited information regarding the characteristics of the TCR repertoire to defined viral antigens in humans. Here, we report a comprehensive analysis of CD8 TCR repertoire diversity (size, V and J gene usage), the consensus binding motifs in CDR3, affinity, and structure of selected a/b paired TCRs to influenza A virus matrix protein M1 (GILGFVFTL, GIL) on HLA-A2+ healthy individuals. Using high-throughput sequencing coupled with the unique molecular identifier (UMI) method, we identified several hundred GIL-binding TCRa and TCRb. TRAV13-1 (63.2%) and TRAJ42 (72.5%) were most frequently found in alpha chain whereas TRBV19 (94.9%) and TRBJ2-5 (54.8%) were frequently found in beta chain. Furthermore, the length of the CDR3 amino acid region was highly restricted with over 70% of alpha (10) and of beta (11). CDR3 sequence analysis revealed shared motifs among different TCRs (a and b chains). We isolated paired expressed TCRa/b from single cells for further structural analysis. The affinity between TCRs and GIL-HLA-A2 complexes showed a wide range (1.8–191 mM). Finally, we resolved two crystal structures of TCRs-GIL-HLA-A2 complexes and found that the TCRs exhibited similar patterns of interactions with the antigen, even though the V, J, and CDR3 lengths were different. Together, our findings revealed the size of the antigen-specific CD8 TCR repertoire, the consensus binding motifs in CDR3, binding affinity of TCR and pMHC, and shared patterns of TCR-pMHC interactions.
Regulation of macrophage anti-microbial functions by the histone methyltransferase MLL1
Authors: Carson William F , Cavassani Karen , Soares Elyara , Hirai Soichiro , Kittan Nicolai , Joshi Amrita , Schaller Matthew , Gallagher Katherine , Matsukawa Akihiro , Aronoff David M , Dou Yali , Kunkel Steven L 
Published: January 2023

Abstract
Mixed-lineage leukemia 1 (MLL1) is a chromatin-modifying enzyme responsible for histone modifications thought to activate gene transcription. Previous studies have linked MLL1-dependent histone methylation to gene expression in response to inflammatory stimuli. To determine if MLL1 plays a role in driving immune responses during inflammatory responses, mice deficient in a single copy of MLL1 (MLL1+/−) were subjected to an experimental model of polymicrobial peritonitis. MLL1+/− mice exhibited increased survival as compared to control animals, correlating with a decreased bacterial burden and decreased levels of proinflammatory chemokines and cytokines in the peritoneal lavage. Bone-marrow derived macrophages from MLL1+/− mice exhibited increased antimicrobial activity in vitro, including increased phagocytosis of bio-particles and increased killing of Group A Streptococcus. Similar results were observed in vitro with MLL1 knockout macrophages (Lys2-Cre MLLfx/fx), with Cre+ bone marrow-derived macrophages exhibiting increased killing of Group A Streptococcus as compared to Cre− littermate-derived macrophages. Gene chip analysis of RNA expression in Cre− and Cre+ macrophages indicated numerous genes whose expression patterns were modulated as a result of MLL1 deficiency in response to classical activation stimuli. Expression pattern changes included both expected decreases in proinflammatory genes, and also surprisingly increases in genes associated with interferon responses and antimicrobial defense. These results implicate MLL1 as a key epigenetic regulator of the complex inflammatory and anti-microbial functions of macrophages.
Cholesterol crystals activate the lectin complement pathway via ficolin-2 and MBL - implications for the progression of atherosclerosis
Authors: Pilely Katrine , Rosbjerg Anne , Genster Ninette , Gál Péter , Pál Gábor , Stahl Gregory L. , Mollnes Tom Eirik , Espevik Terje , Garred Peter 
Published: January 2023

Abstract
Cholesterol crystals (CC) play an essential role in the formation of atherosclerotic plaques by inducing inflammation and by functioning as an endogenous danger signal. CC activate the classical and the alternative complement pathways, but the role of the lectin pathway is unknown. In this study we hypothesized that pattern recognition molecules (PRM) from the lectin pathway bind CC and functions as an upstream innate inflammatory signal in the pathophysiology of atherosclerosis. We investigated the binding of the PRMs mannose-binding lectin (MBL), ficolin-1, ficolin-2, and ficolin-3, the associated serine proteases, and complement activation products to CC using recombinant proteins, specific inhibitors as well as deficient and normal sera. Binding was assessed by flow cytometry and microscopy. The results showed that the lectin pathway was activated on CC by binding of ficolin-2 and MBL, resulting in activation and deposition of complement activation products. MBL bound to CC in a calcium dependent manner while ficolin-2 binding was calcium independent. No binding was observed for ficolin-1 or ficolin-3. Moreover, we demonstrated that IgM, but not IgG bound to CC and that C1q binding was facilitated by IgM. In conclusion our study demonstrates that PRMs from the lectin pathway recognize CC and provides evidence for an important role for this pathway in the inflammatory response induced by CC in the pathophysiology of atherosclerosis.
The Effect of Hepsin in Interferon Induction Pathways
Authors: Liu Helene Minyi , Hsin Fu 
Published: January 2023

Abstract
The induction of type I interferon (IFN) is critical for antiviral innate immune response. During virus infection, Retinoic acid-inducible gene I (RIG-I) recognizes the viral RNA and activates Mitochondrial antiviral-signaling protein (MAVS). MAVS further triggers downstream signaling to activate Interferon regulatory factor 3 (IRF3) to induce type I IFN. Many viral proteases cleave adapter proteins in the IFN induction pathway, such as HCV NS3/4A cleaving MAVS, as a strategy to evade anti-viral innate immunity. However, it is not clear whether host protease may also regulate IFN induction pathway to control the steadily low expression of IFN in uninfected cells. Here we report that Hepsin (HPN), which belongs to type II transmembrane serine protease family, could inhibit the production of type I IFN during RNA virus infections in several cell-lines. Hepsin expression could be detected in Huh7 and HepG2 hepatocytes but not ubiquitously in all cell types. Knocking-out Hepsin in mouse embryonic fibroblasts resulted in higher expression of IFNβ and interferon response genes (ISGs). Besides, when co-transfected with N-RIG, which is constitutively activating IFNβ expression, Hepsin could decrease IFNβ promoter activity. However, under the expression of IRF3-5D, Hepsin could not reduce IFNβ promoter activity, suggesting that Hepsin might target certain adaptor proteins downstream of RIG-I and upstream of IRF3. We showed an inverse correlation with wild-type Hepsin expression but not the protease-deficient mutant Hepsin in the IFNβ promoter activities . Consequently, these results reveal a novel role of Hepsin in regulating the type I IFN induction pathway in hepatocytes.
Emerging roles of an innate immune regulator TAPE in Toll-like receptors, RIG-I-like receptors, and beyond
Authors: LING PIN , Chen Kuan-Ru , Kao Chen-Chu , Chuang Huai-Chia , Tan Tse-Hua 
Published: January 2023

Abstract
Pattern-recognition receptors (PRRs) trigger innate immune defenses against pathogen infection via downstream signaling pathways linking to inflammation and cell-autonomous immunity like phagocytosis and autophagy. IKK family kinases, IKKα and IKKβ, function to relay PRR signals to proinflammatory cytokine production to amplify innate immune responses. TBK1, a non-canonical IKK kinase, links nucleic acid sensors to type I interferon induction against viral infection and also regulates the autophagic clearance of intracellular bacteria. TBK1-Associated Protein in Endolysosomes designated TAPE, also known as CC2D1A, is an innate immune regulator acting upstream of Trif to regulate the TLR3 and TLR4 pathways, or upstream of MAVS to regulate the cytosolic RIG-I-like receptor (RLR) pathways. To our best knowledge, TAPE is the first regulator implicated in both the endosomal TLR and cytosolic RLR pathways at such an early step. We are thus interested in investigating in vivo roles of TAPE in innate immunity and molecular mechanisms by which TAPE regulates TLRs, RLRs, and possibly other PRRs. TAPE conditional knockout (cKO) mice, in which TAPE is selectively disrupted in immune cells, have been generated for our study. Our results showed that upon influenza A virus infection, TAPE cKO mice exhibited a more severe mortality than wild type mice. Further, TAPE cKO mice were shown to be more susceptible to Salmonella Typhimurium infection but more resistant to LPS-induced septic shock. Notably, ex vivo results showed that TAPE was critical for the autophagic clearance of Salmonella Typhimurium. Together, our data support a critical role for TAPE in regulating innate immune defenses through TLRs, RLRs, and autophagy.
EHD1 as a Positive Regulator of Macrophage CSF1R
Authors: Cypher Luke R , Bielecki Timothy Alan , Iseka Fany M , Band Vimla , Band Hamid 
Published: January 2023

Abstract
Macrophage biology is regulated via receptor tyrosine kinase (RTK) CSF-1 receptor (CSF1R). Anomalous macrophage biology has been shown to be hallmark in diseases such as cancer and atherosclerosis. Although CSF1R signaling has been studied, little is known about the mechanisms which govern the endocytic trafficking of the receptor. EHD1 is known to regulate endocytic recycling of receptors, but the role of EHD1 in macrophages is unstudied. In this study, we discovered EHD1 knockout (KO) murine bone marrow-derived macrophages (BMDMs) had markedly reduced total and surface CSF1R levels and a corresponding decreased downstream functional output due to decreased CSF1R signaling. EHD1 KO led to perinuclear CSF1R accumulation which co-localized with the Golgi (GM130). These findings reveal a novel and essential role for EHD1 in transporting newly synthesized CSF1R to the cell surface. Our study expands our understanding of RTK trafficking which has potentially dramatic implications for designing new targeted therapies for devastating human diseases.
The C-terminal tail of TRIM56 dictates antiviral restriction of influenza A and B viruses by impeding viral RNA synthesis
Authors: Liu Baoming , Li Nan L , Shen Yang , Bao Xiaoyong , Fabrizio Thomas , Elbahesh Husni , Webby Richard J. , Li Kui 
Published: January 2023

Abstract
Accumulating data suggest that tripartite motif-containing (TRIM) proteins participate in host responses to viral infections, either by acting as direct antiviral restriction factors or through regulating innate immune signaling of the host. Of &gt;70 TRIMs, TRIM56 is a restriction factor of several positive-strand RNA viruses, including three members of the family Flaviviridae (yellow fever virus, dengue virus and bovine viral diarrhea virus) and a human coronavirus (OC43), and this ability invariably depends upon the E3 ligase activity of TRIM56. However, the impact of TRIM56 on negative-strand RNA viruses remains unclear. Herein, we show TRIM56 puts a check on replication of influenza A and B viruses in cell culture, but does not inhibit Sendai virus or human Metapneumovirus, two paramyxoviruses. Interestingly, the anti-influenza activity was independent of the E3 ligase activity, B-box, or coiled-coil domains. Rather, deletion of a 63-residue long, C-terminal tail portion of TRIM56 abrogated the antiviral function. Moreover, expression of this short C-terminal segment curtailed the replication of influenza viruses as effectively as that of full-length TRIM56. Mechanistically, TRIM56 was found to specifically impede intracellular influenza virus RNA synthesis. Consistent with this, a fraction of TRIM56 was re-distributed to the nucleus after IAV infection. Together, these data reveal novel antiviral activity of TRIM56 against influenza A and B viruses and provide novel insights into the mechanism by which TRIM56 restricts these medically important orthomyxoviruses. These findings raise the possibility that small molecules mimicking the C-terminal portion of TRIM56 could be developed into broad influenza antivirals.
Plasmid electrotransfer activates DNA pattern recognition receptors and induces production of proinflammatory molecules
Authors: Heller Loree , Bosnjak Masa , Znidar Katarina , Cemazar Maja 
Published: January 2023

Abstract
Electroporation or electrotransfer is an effective delivery system for introducing plasmid DNA into cells and tissues. During this process, pDNA primarily enters the cell via endocytosis, although some DNA reaches the cytosol. Endosomal, cytosolic, and nuclear DNA-specific pattern recognition receptors (PRRs) in many cell types have been described. Activated PRRs induce the production of proinflammatory molecules and programmed cell death.
In B16.F10 mouse melanomas, electrotransfer of empty backbone plasmid induced the production of several proinflammatory cytokines and chemokines including IFNβ, implicating the binding and activation of intracellular DNA-specific PRRs. Histologically, tumor necrosis independent of caspase-3 was observed. The purpose of this study was to investigate whether tumors and cells express cytosolic DNA sensors and whether these sensors respond to pDNA electrotransfer.
Although the mRNAs for several DNA sensors were detected in tumors, none was significantly upregulated. In B16.F10 cells in culture, IFNβ mRNA and protein levels were significantly upregulated after DNA electrotransfer. The mRNAs for several DNA sensors were present in these cells and DAI, DDX60, and p204 mRNAs were significantly upregulated after pDNA electrotransfer. DDX60 protein levels were coordinately upregulated. Mirroring the observation of tumor necrosis, cells underwent a significant DNA concentration-dependent decrease in proliferation and survival.
Taken together, increased IFNβ and DNA sensor expression accompanied by cell death and tumor necrosis indicate that DNA electrotransfer activates intracellular DNA sensors in B16.F10 cells and tumors, producing both in vitro and in vivo effects.
CoCr microparticles induce a robust type 2 innate immune response that is dependent on SYK and BTK signaling
Authors: Palma Mark J , Mishra Pankaj Kumar , Millman Ariel , Hallab Nadim J , Beebe Kathleen , Benevenia Joseph , Gause William C 
Published: January 2023

Abstract
Cobalt chrome (CoCr) is a major component of orthopaedic implants and a source of metallic wear debris, which may contribute to harmful inflammation and implant failure. Intra-peritoneal inoculation of CoCr induces a robust innate type 2 response with increases in M2 macrophages, eosinophils, and neutrophils and this response is dependent on Spleen tyrosine kinase (SYK) and Bruton’s tyrosine kinase (BTK) signaling, but not TLR, Caspase-1, or Cyclooxygenase signaling. We have also shown that this robust response is independent of T and B cell signaling. To examine this response in a more clinically relevant murine model, bilateral intra-articular knee injections of CoCr microparticles were given to C57BL/6 mice and changes in different innate immune cells were assessed from knee synovial cell isolates. Administration of the SYK inhibitor BAY 61-3606, or the BTK inhibitor Ibrutinib, abrogated innate immune cell recruitment and M2 macrophage polarization which is otherwise observed 2 days after inoculation with CoCr microparticles. The presence of M2 macrophages (CD68+, CD206+, and CD163+) is also observed in human peri-prosthetic tissue samples obtained from patients undergoing revision joint arthroplasty. Taken together, our findings suggest that BTK signaling is required for the type 2 inflammatory response to CoCr microparticles, providing a potential novel target for control of harmful inflammation leading to fibrosis and aseptic loosening.