bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparative analysis of signature genes in Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)-infected porcine monocyte-derived dendritic cells at differential activation statuses.
Authors: Miller Laura C , Sang Yongming , Bayles Darrell O , Blecha Frank 
Published: January 2023

Abstract
Activation statuses of monocytic cells, e.g. monocytes, macrophages and dendritic cells (DCs), are critically important for antiviral immunity. In particular, some devastating viruses, including porcine reproductive and respiratory syndrome virus (PRRSV), are capable of directly infecting these cells to subvert host immunity. Monocyte-derived DCs (mDCs) are major target cells in PRRSV pathogenesis; however, the plasticity of mDCs in response to activation stimuli and PRRSV infection remains unstudied. In this study, we polarized mDCs, and applied genome-wide transcriptomic analysis to compare signature genes involved in mDCs activation and response to PRRSV infection. Our long-term goal is to integrate activation status with antiviral responses in these cells and to functionally modulate them for a prototypic cellular adjuvant/vaccine that is ideal for potentiating antiviral immunity. Porcine mDCs were polarized with mediators for 30 hours, then mock-infected, infected with PRRSV strain VR2332, or a highly pathogenic PRRSV strain (rJXwn06), for 5 h. Total RNA was extracted and used to construct sequencing libraries for RNA-Seq procedures previously optimized. Comparisons were made between each polarized and unpolarized group (i.e. mediator vs. PBS), and between PRRSV-infected and uninfected cells stimulated with the same mediator. Many of the genes showing the most variability were related to cellular structure and innate immune response. The magnitude of differentially expressed gene profiles detected in HP-PRRSV rJXwn06 infected mDCs as compared to VR-2332 infected mDCs was consistent with the increased pathogenicity of the HP-PRRSV in vivo.
Transcript analysis of natural killer (NK) cell specific genes in the liver, kidney and spleen tissues of rag1
−/− mutant zebrafish in response to in vivo administration of TLR ligands
Authors: Muire Preeti Judith , Hanson Larry , Yoder Jeffrey , Petrie-Hanson Lora 
Published: January 2023

Abstract
T and B cells mediate specific protective immunity. NK cells can also mediate a memory response to murine cytomegalovirus in T and B cell deficient rag2−/− mice. T and B cell deficient rag1−/− mutant zebrafish mediate protective immunity to intracellular bacteria. NK cells have not been characterized in zebrafish due to a lack of tools to target specific markers. The aim of this study was to evaluate NK cell tissue distribution and transcript response to TLR ligands in rag1−/− mutant zebrafish. We accomplished this by determining the level of expression of ifnγ, t-bet, nitr9, and NK lysin (nkl) a, b, c and d transcripts following exposure to β glucan, Poly I:C, R-848 and VTX-2337. Expression levels were normalized to the basal level of expression and the fold changes were compared to that of endotoxin free PBS (carrier control) injected group. R848 and VTX-2337 exposure changed the gene expression profile in a tissue specific manner. Nitr9 protein expression was analyzed by western blot and detectable levels were found in the liver. Nitr9 expression correlated with the expression of ifnγ in liver and kidney. In another experiment, fish were injected with TLR ligand when vaccinated, were only vaccinated, or not vaccinated. After bacteria challenge, fish that received β glucan or R848 when vaccinated had the highest survival. This suggests the TLR ligands have an adjuvant activity on NK cell protective responses. Our results of NK cell stimulation indirectly demonstrate the presence of NK cells in zebrafish and suggest a possible means of enhancing NK cell based immunity. These findings demonstrate the clinical significance of the rag1−/− mutant zebrafish model.
Functional impairment of CMV-reactive cellular immunity during pregnancy
Authors: Boeckl Katharina , Reuschel Edith , Barabas Sascha , Bendfeldt Hanna , Deml Ludwig , Seelbach-Göbel Birgit 
Published: January 2023

Abstract

Background
Cytomegalovirus (CMV) is the most common congenital viral infection. Mother-to child transmission (MTCT) can cause severe disability of the child. Intact CMV-specific cell-mediated immunity (CMI) has been shown to prevent uncontrolled replication in healthy individuals. In this study we aim to determine whether CMV-specific CMI is impaired in pregnant women, thus increasing the overall risk for active CMV replication and transmission.

Methods
CMV-specific CMI in peripheral blood of 61 pregnant women in the three trimenons of their pregnancy has been determined using T-Track® CMV for detection of IE-1 and pp65-reactive effector cells by IFN-γ ELISpot. Additionally CMV- IgG and IgM titers were examined.

Results
CMV-specific CMI was detected in 70 % of CMV-seropositive pregnant women. Also 4 % of CMV-IgG seronegative women showed IE-1- but not pp65- reactive cells. The overall number of CMV-reactive cells in pregnant women was lower compared to a matched non-pregnant control group. Postpartum the IE-1- but not pp65- specific CMI is still lower than in the non-pregnant control group.

Conclusion
Functional analysis of CMV-reactive immune cells using T-Track® CMV indicates a systemic down-regulation of the CMV-specific CMI in pregnant women.
The IFIH1 A946T autoimmune risk variant exhibits enhanced signaling and promotes T1D pathogenesis
Authors: Gorman Jacquelyn A , Hundhausen Christian , Errett John , Ge Yan , Arkatkar Tanvi , Clough Courtnee , Dai Xuezhi , Khim Socheath , Oukka Mohamed , Concannon Patrick , Gale Michael , Buckner Jane H , Rawlings David J 
Published: January 2023

Abstract
Genome wide association studies have linked SNPs in innate signaling programs to an increased risk for autoimmune disease. One SNP, rs1990760, within the IFIH1 viral sensor (leading to an A to T amino acid change at codon 946) is strongly associated with multiple autoimmune diseases including type 1 diabetes (T1D). The impact of this variant on viral sensing and autoimmune pathogenesis, however, has not yet been determined. Using an overexpression system, we demonstrate that IFIH1-A946T exhibits heightened basal and post-stimulation signaling and increased viral control. Consistent with this finding, primary PBMCs from healthy subjects expressing the variant exhibit heightened signaling. To study IFIH1-A946T in vivo, we generated a novel knock-in murine model. Both heterozygous and homozygous variant mice display enhanced basal Type I IFN mRNA expression and splenomegaly. Moreover, homozygous pups manifest an embryonic survival defect. Further, IFIH1-T946 mice exhibit increased frequency of STZ-induced T1D. Finally, intercross of IFIH1-T946 mice with animals expressing the T1D risk variant in PTPN22 lead to a synergistic increase in T1D development. Together, our data demonstrate that IFIH1-A946T alters antiviral signaling and strongly supports the model that increased Type I IFN production driven by the variant leads to an inflammatory state that promotes autoimmunity, particularly in association with other autoimmune risk alleles.
TAPP adaptors control Akt-dependent metabolic activation of germinal center B cells and suppress autoimmunity via binding to the SHIP product PI(3,4)P2
Authors: Marshall Aaron J , Jayachandran Nipun , Landego Ivan , Hou Sen , Meija Edgard M , Sheikholeslami Kimia , Hatch Grant M 
Published: January 2023

Abstract
The phosphoinositide phosphatase SHIP regulates B cell activation by converting the PI 3-kinase product PI(3,4,5)P3 to PI(3,4)P2. While this activity of SHIP appears critical for generation of effective adaptive immunity while avoiding autoimmunity, the function of PI(3,4)P2 in B cells remains relatively unexplored. Tandem PH domain containing proteins (TAPPs) are adaptor proteins that specifically bind to PI(3,4)P2 via their C-terminal PH domains, targeting them to the plasma membrane. Mice bearing inactivating mutations in the PH domains of both TAPP1 and TAPP2, uncoupling them from PI(3,4)P2, exhibit hypergammaglobulinemia and develop lupus-like characteristics including anti-DNA antibodies and deposition of immune complexes in kidneys. Here we find that TAPP KI mice develop chronic germinal centres (GCs) and age-associated increases in B cell expression of activation, exhaustion and memory markers, all of which are B cell intrinsic and dependent on the co-stimulatory molecule ICOS. TAPP KI B cells exhibit elevated phosphorylation of Akt and its target GSK3, and increased survival. Strikingly, TAPP KI B cells exhibit increased metabolic activity associated with elevated oxygen consumption rate, increased extracellular acidification rate, increased expression of glucose transporter Glut1 and increased glucose transport rate. Together our findings suggest that TAPP-PI(3,4)P2 interaction is important for regulating signalling via Akt, B cell metabolism and development of autoimmunity.
Apoptotic nucleolus is recognized by complement C1q and degraded by C1 proteases
Authors: Lu Jinhua , Cai Yitian , Teo Boon Heng Dennis , Yeo Joo Guan , Chen Junjie 
Published: January 2023

Abstract
In infections, complement C1q recognizes pathogen-congregated antibodies and elicits complement activation. Among endogenous ligands, C1q binds to DNA and apoptotic cells but whether C1q binds to nuclear DNA in apoptotic cells remains to be investigated. With UV-induced apoptosis, C1q initially bound to peripheral cellular regions in early apoptotic cells. By 6 hr, binding concentrated in the nuclei to the nucleolus but not the chromatins. When nucleoli were isolated from non-apoptotic cells, C1q also bound to these structures. By affinity chromatography, a number of nuclear proteins were found to bind to C1q including nucleophosmin 1 and nucleolin. In vivo, C1q exists as C1 complex (C1qC1r2C1s2) and C1q binding to ligands activates the C1r/C1s proteases. Incubation of nucleoli with C1 caused degradation of nucleolar proteins neucleolin and nucleophosmin 1. Cleavage of purified nucleophosin 1 by C1 is studied in details revealing three defined fragments of 11, 17 and 18 kDa, respectively. This was inhibited by C1 inhibitor. The nucleoli are abundant with autoantigens. C1q binding and C1r/C1s degradation of nucleolar antigens during cell apoptosis potentially reduce autoimmunity. These findings suggest broader substrate spectrum of C1 proteases and help understand why genetic C1q and C1r/C1s deficiencies cause systemic lupus erythematosus.
IL-17 drives but can also restrain psoriatic inflammation via distinct, target cell-specific mechanisms
Authors: Siebenlist Ulrich , Ha Hye-lin , Wang Hongshan , Claudio Estefania , Tang Wanhu 
Published: January 2023

Abstract
Psoriasis is a chronic inflammatory skin disease characterized by abnormal keratinocyte proliferation and differentiation and by an influx of inflammatory cells. The mechanisms underlying psoriasis in humans and mice are still only partially understood, although IL-17 cytokines are known to make crucial contributions, cytokines that signal via the obligatory adaptor CIKS/Act1. Here we identify critical roles of CIKS/Act1-mediated signaling in imiquimod-induced psoriatic inflammation, a mouse model that shares features with the human disease. We found that IL-17 cytokines/CIKS-mediated signaling into keratinocytes was essential for neutrophilic microabscess formation; in addition, it contributed to hyperproliferation and markedly attenuated differentiation of keratinocytes, at least in part via direct effects. Interestingly, IL-17 signaling into keratinocytes also exerted a negative feedback to eventually delimit further accumulation of IL-17-producing γδ T cells in skin. This stands in contrast to IL-17 cytokines/CIKS-mediated signaling into non-keratinocytes, particularly dermal fibroblasts, which promoted cellular infiltration and enhanced accumulation of IL-17-producing γδ T cells in skin, comprising an initial positive feed-forward mechanism. We will report on recent progress in understanding in particular the mechanism by which signaling into keratinocytes eventually feeds back to prevent excessive IL-17 production. Our data show that CIKS-mediated signaling is dynamic and central to the development of both dermal and epidermal hallmarks of psoriasis, but also activates mechanisms that delimit inflammation.
BCAP inhibits myeloid cell development from hematopoietic progenitors.
Authors: Duggan Jeffrey M , Hamerman Jessica A 
Published: January 2023

Abstract
B cell adaptor for PI3-kinase (BCAP) is a signaling adaptor expressed in hematopoietic cells including macrophages, monocytes, and neutrophils. Here we asked if BCAP plays a role in development of these myeloid cells. We found that BCAP was expressed in bone marrow (BM) hematopoietic progenitors, including LSK (Lin−Sca1+cKit+), CMP (Common Myeloid Progenitor) and GMP (Granulocyte/Macrophage Progenitor) cells, suggesting that BCAP may impact myelopoiesis. BCAP−/− mice had more BM monocytes than WT mice, and in mixed chimeras generated with a 1:1 ratio of WT and BCAP−/− BM, monocytes and neutrophils in the BM, blood and spleen exhibited skewing towards BCAP−/− origin, showing a competitive advantage for BCAP−/− myeloid cells. Thus we hypothesized that BCAP inhibits myeloid development. Consistent with this hypothesis, BCAP-deficient BM LSK, CMP and GMP cells out-competed WT progenitors in mixed chimeras. In an in vitro myeloid colony-forming-unit assay, sorted BCAP−/− progenitors produced more myeloid cells than WT progenitors, supporting a cell-intrinsic role of BCAP in inhibiting myeloid differentiation. Furthermore, BCAP−/− progenitors were more mature than WT progenitors, indicating that BCAP−/− progenitors display accelerated myeloid development. During cyclophosphamide-induced myeloablation or specific monocyte depletion, BCAP−/− mice replenish circulating monocytes earlier than WT mice. Lastly, BCAP−/− progenitors showed increased expression of the myeloid-differentiating transcription factors IRF8 and C/EBPα compared to WT progenitors. Together, these data identify BCAP as an inhibitor of myeloid development from BM hematopoietic progenitors in the steady state and during demand conditions.
Propranolol modulates androgen-expanded myeloid-derived suppressor cells (MDSC) in severely burned mice
Authors: Kobayashi Makiko , Ito Ichiaki , McCalla Claudia T , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
Severe burn injuries produce chronic hypermetabolic and hypercatabolic states persisting long after injury. For the improvement of net protein balance and lean body mass, we are trying to use oxandrolone (OX, a synthetic analog of testosterone) in severely burned patients subjected to the standard of care (SOC) including propranolol (PPL, a blocker of β-adrenergic receptors). OX has the potential to exacerbate post-burn immunoparalysis through the expansion of MDSC (Gr-1+CD11b+ cells). Since propranolol accelerates MDSC maturation, the effect of OX on the expansion of MDSC in severely burned mice subjected to SOC was studied. Mice were treated s.c. with OX (0.1 mg/kg, 1 and 12 h post burn) alone or in combination with PPL (3.3 mg/kg, 12 h post burn). MDSC in splenic MNC from these mice were analyzed by flow cytometry. Also, peritoneal Mϕ from these were analyzed for intracellular TNF-α, as an inhibitor of MDSC maturation. In the results, the number of MDSC increased 5.5-fold in burn mice treated with OX, as compared to that of MDSC in burned mice. However, this increase was not seen in burn mice treated with OX and PPL in combination. TNF-α+ cells increased 2.5-fold in the peritoneal Mϕ of burn mice treated with OX, but not in burn mice treated with OX and PPL. These results show that the number of MDSC is not increased by OX in severely burned mice treated with PPL.
NF-κB restricts NLRP3 inflammasome activation via p62-dependent mitophagy which eliminates mitochondrial signals
Authors: Zhong Zhenyu , Umemura Atsushi , Sanchez-Lopez Elsa , Liang Shuang , Shalapour Shabnam , Wong Jerry , He Feng , Boassa Daniela , Perkins Guy , Ali Syed Raza , McGeough Matthew G , Ellisman Mark H , Seki Ekihiro , Gustafsson Asa B , Hoffman Hal M , Diaz-Meco Maria T , Moscat Jorge , Karin Michael 
Published: January 2023

Abstract
NF-κB, a key activator of inflammation primes the NLRP3-inflammasome for activation by inducing pro-IL-1β and NLRP3 expression. NF-κB, however, also prevents excessive inflammation and restrains NLRP3-inflammasome activation through a poorly defined mechanism. We now show that NF-κB exerts its anti-inflammatory activity by inducing delayed accumulation of the autophagy receptor p62/SQSTM1. External NLRP3-activating stimuli trigger a form of mitochondrial (mt) damage that is caspase-1- and NLRP3-independent and causes release of direct NLRP3-inflammasome activators, including mtDNA and mtROS. Damaged mitochondria undergo Parkin-dependent ubiquitin conjugation and are specifically recognized by p62, which induces their mitophagic clearance. Macrophage-specific p62 ablation causes pronounced accumulation of damaged mitochondria and excessive IL-1β-dependent inflammation and enhances macrophage death. Therefore, the “NF-κB-p62-mitophagy” pathway is a macrophage-intrinsic regulatory loop through which NF-κB restrains its own inflammation-promoting activity and orchestrates a self-limiting host response that maintains homeostasis and favors tissue repair. This pathway, however, is bypassed when NLRP3 is genetically activated.
Elucidation of the NLRP3 interactome
Authors: Fairgrieve Marian , Davis Michael , Gale Michael J 
Published: January 2023

Abstract
Upon pathogen detection, the immune system triggers a variety of signaling pathways. Two key pathways are the proinflammatory NLRP3 inflammasome pathway, which activates the proinflammatory cytokines IL-1β and IL-18 via activation of Caspase-1 (CASP1), and the antiviral IFN pathway, which signals through the JAK-STAT pathway to upregulate key IFN-stimulated genes. Importantly, a growing body of evidence demonstrates that crosstalk between these pathways is required to fine tune the host immune response, and the co-regulation between the pathways can be either positive or negative. For example, upon viral infection, IL-1β synergizes with INF to suppress viral replication. At other times, as in the treatment of multiple sclerosis with INF-β therapy, INF suppresses IL-1β expression and activation. However, the mechanism governing inflammasome-INF crosstalk remains unclear. To address this gap in our understanding, we conducted a series of proteomic studies aimed at identifying novel interacting partners of NLRP3 and CASP1. We employed an affinity purification and mass spectrometry-based approach. Interestingly, we found that components of the JAK-STAT pathway interact with NLRP3. We will present data characterizing this interaction as well as the role of JAK-STAT proteins in regulating inflammasome-IFN crosstalk. These findings will enhance our understanding of how these two cascades act in concert to impact infection and immunity.
Mutant Interferon-γ receptor 1 lacking cytoplasmic domain may transduces signals leading to mycobacterial susceptibility
Authors: Shieh Chi-Chang K , Chiu Darren Jianjhih , Chiang Kuo-Ming , Yen C-L 
Published: January 2023

Abstract
Interferon-γ (IFN-γ) is a crucial cytokine for innate and acquired immunity against infections by intracellular bacteria including mycobacteria. Mutations in the genes involved in the IL-12-IFN-γ pathway cause impaired immunity against lowly virulent mycobacteria and lead to Mendelian Susceptibility to Mycobacterial Disease (MSMD). How the mutations differentially affect leukocyte activation signals and lead to different clinical presentations has not been characterized. We previously analyzed a family with autosomal dominant MSMD and found an 818del4 mutation on the IFN-γ receptor gene IFNGR1 leading to high expression of IFN-γ receptor on leukocytes. While IFN-γ-induced IL-12 and TNF-α secretion from peripheral blood mononuclear cells (PBMCs) were low in the patients. IFN-γ and LPS-induced NADPH oxidase priming and hence reactive oxygen species production was normal. We hence cloned the 818del4 mutant and wildtype IFNGR1 into lentiviral vector and expressed the mutant IFNGR1 on human embryonic kidney 293T cells. We found that stimulation with IFN-γ increased the phosphorylation of Akt and activated p47phox through phosphorylation of Erk1/2 in cells expressing 818del4 mutant IFNGR1. Our results implicated that the truncated IFN-γ receptor may transmit differential signals in leukocytes which alter the toll-like receptor signals by activating the Akt activity and contribute to the severity of immunodeficiency in patients with autosomal dominant MSMD.
Amphibian innate immune mediators protect against human Influenza strains
Authors: Holthausen David J , George Sanil , Jacob Joshy 
Published: January 2023

Abstract
Frogs and toads are incredible reservoirs of biologically active peptides. Amphibians secrete host defense peptides from their skin as part of their innate immune response. This ancient response acts to protect the amphibians against microbes. The quantity and scope of these secreted peptides dwarfs mammalian analogues, accounting for a substantial portion of all known host defense peptides. The non-invasive and non-harmful methods for frog peptide collection, in tandem with the abundance and breadth of these peptides, makes them excellent choices for novel peptide drug therapies. Studies have shown that these peptides can effectively neutralize enveloped viruses, mycobacteria, gram-negative and gram-positive bacteria, fungi, and even cancerous or transformed cells. Because of the nature of antimicrobial peptides, often targeting the most critical, conserved aspects of a micro-organism, they may prove a vital alternative to conventional drugs plagued by pathogen resistance. Given this untapped potential for anti-viral therapies, we assessed novel host defense peptides from the skin of the Indian fungoid frog, Hylarana malabarica. During our analysis, we isolated several peptides from H. malabarica that show anti-viral activity against human influenza viruses. Our studies indicate that peptides from H. malabarica demonstrate anti-influenza activity in vitro, and also show potential as an anti-viral therapy in vivo.