bluuebunny/crossref_metadata_2025_split huggingface dataset:
Defining how mitochondrial DNA stress regulates innate immune responses
Authors: Martinez Camila Guerra , Lei Yuanjiu , West Phillip 
Published: January 2023

Abstract
Polymerase gamma (pol γ) is nuclear-encoded protein responsible for mtDNA replication. POLG mutations are recognized as a significant cause of primary mitochondrial diseases and are implicated in a range of disorders. Several mouse models of POLG-related disease have been reported, the most well studied is the POLG-mutator mouse. Animals homozygous for the mutant alleles exhibit elevated mtDNA instability, and POLG-mutator mice exhibit pathology that mirrors various aspects of human POLG-related diseases. The literature has few descriptions about the immune system of POLG-mutator mice or patients with POLG-related disorders, despite the fact that aberrant immune responses have been implicated in the pathology of diseases with similar phenotypes. We have found that mitochondrial dysfunction, resulting from POLG mutation, alters innate immune responses at baseline and after challenge, which we hypothesize may exacerbate pathology in POLG-related mitochondrial disorders. To explore this hypothesis, we have stimulated macrophages from WT and POLG-mutator mice with various innate immune stimuli, and have uncovered that POLG-mutator macrophages display alterations in type I interferon responses, but strikingly, exhibit profound decreases in inflammasome-dependent interleukin-1 beta (IL-1b) secretion. Ongoing work will determine how mitochondrial dysfunction impairs inflammasome activation and IL-1b secretion in order to appreciate how aberrant innate immune responses influence POLG-related disease progression.
LRRK2 is required for macrophage homeostasis and the control of type I IFN
Authors: Weindel Chi Gidley , Bell Samantha L. , Vail Krystal , West A. Phillip , Patrick Kristin L. , Watson Robert O. 
Published: January 2023

Abstract
The use of genetic association studies has helped us appreciate that inflammation and immune dysfunction play an integral role in the pathology of multiple chronic disorders including cancer, type II diabetes, and neurodegenerative disease. We became interested in Leucine-rich repeat kinase 2 (LRRK2), a multifunctional kinase associated with Parkinson’s disease (PD), when it was revealed that LRRK2 polymorphisms are also associated with bacterial susceptibility to Mycobacterium leprae, a close relative of Mycobacterium tuberculosis, both of whose pathology relies heavily upon type I interferon (IFN), an innate immune response that activates effector cells. The regulation of type I IFN is critical to prevent a systemic inflammatory state and for the maintenance of healthy neurons. We therefore hypothesized that LRRK2 has a role in regulating innate immunity, contributing to both PD pathogenesis and bacterial infection. We found that in the absence of LRRK2 (Lrrk2−/−), macrophages failed to produce type I IFN in response to multiple cellular insults including, bacterial infection, mitochondrial damage, and DNA/RNA transfection. Interestingly the defect in type I IFN was a result of an imbalance at the level of the mitochondria, which altered macrophage homeostasis. These data suggest that LRRK2 is critical component of the innate immune system through its role maintaining mitochondrial health. The fact that LRRK2-deficient cells cannot induce type I IFN has major implications on both the maintenance of a proper cytokine milieu in the brain and management of the body’s immune response to infection.
Targeting of Diffuse Large B-cell Lymphomas using MyD88 small molecule inhibitors
Authors: Snyder Greg A. , Brown Lindsey J. , Taylor Savannah , Faupel Ciara E. , Sherman Matthew , Montague Justin J. , Saikh Kamal , Wang Yajing 
Published: January 2023

Abstract
A mutation associated with nearly 1/3 of human diffuse large B cell lymphomas (DLBCLs) has been identified within MyD88. This mutation correlates with tumor cell proliferation and survival involving spontaneous and sustained activation of NF-κB signaling. MyD88 is a central signaling adapter for the Interleukin-1 (IL-1R) and Toll-like receptors (TLRs). In normal healthy cells, MyD88 is thought to be held in an auto-inhibitory state with its own death and TIR domains fused together in negative self-regulation until activated by appropriate receptor mediated ligand engagement.

Observation
CADD derived small molecule compounds inhibit MyD88 dimer formation and protect against Staphylococcal enterotoxin B (SEB) induced death in animal models.

Hypothesis
Based on this observation we hypothesize that MyD88 specific small molecule inhibitors may be useful in treating DLBCLs bearing MyD88L265P.

Approach
Using in vitro and in vivo studies we characterize MyD88 specific SMIs for the ability to inhibit tumor cell proliferation and signaling in cancer cells bearing the oncogenic mutation MyD88L265P.

Results
1) MyD88 SMIs are able to inhibit cell proliferation of DLBCLs bearing the MyD88L265P as measured by MTS cell proliferation assay, 2) recombinant MyD88 and SMIs exhibit unique binding chromatograms in comparison to DMSO controls as measured by thermal shift assay and 3) MyD88 SMIs are partially able to inhibit LPS activated TLR4 cell NF-kB signaling in comparison to the TLR4 specific inhibitor TAK242. Future studies defining the molecular mechanism of this mutation with additional human patient tumor isolates will inform and propel development of novel therapeutics to counteract both inflammation as well as tumor formation.
CARD19: a mitochondrial caspase recruitment domain protein with a role in regulating pro-inflammatory innate immune responses
Authors: Rios Kariana Elizabeth , Liem Ayesha , Kashyap Anuj , Maynard Sean , Paul Suman , Schaefer Brian C. 
Published: January 2023

Abstract
CARD19 is a ubiquitously expressed protein of unknown function. This protein was originally named BinCARD, and early data suggested that it interacts with Bcl10 and inhibits NF-κB activation in T cells. However, these early data were based on overexpression studies, and we are aware of no in vivo data that support this proposed function. Additionally, the cellular localization of CARD19 remains uncharacterized. We sought to address these gaps in knowledge by examining macrophages, thymocytes, and splenocytes from Card19 +/+ and Card19 −/− mice using superresolution microscopy and flow cytometry; we also compared secretion of TNF-alpha, IL-6, and MCP-1 in Card19 +/+ and Card19 −/− mice injected with LPS. Superresolution microscopy revealed that CARD19 is localized to the outer mitochondrial membrane, with no evidence of co-localization with BCL10. Deletion of the predicted transmembrane domain resulted in nuclear and cytosolic distribution, while a partial deletion of the C-terminus resulted in localization in the endoplasmic reticulum. Data to date do not indicate significant differences between Card19 +/+ and Card19 −/− thymocytes and splenocytes in development, activation, or effector function. Interestingly, Card19 −/− mice displayed higher plasma concentrations of TNF-alpha, IL-6, and MCP-1 relative to Card19 +/+ mice in response to LPS. We conclude that CARD19 does not measurably impact BCL10 function in vivo, and we suggest that CARD19 is likely involved in regulation of a mitochondrial function that affects pro-inflammatory innate immune responses via a mechanism independent of BCL10.
Maternal immune activation induced by lipopolysaccharide triggers response inflammatory, oxidative damage and damage to blood brain barrier integrity and placental barrier in Wistar rats pregnant and fetus
Authors: Simões Lutiana Roque , Generoso Jaqueline S. , Sangiogo Gustavo , Faller Cristiano Julio , Réus Gislaine Zilli , Zugno Alexandra Ioppi , Petronilho Fabricia , Scaini Giselli , Giridharan Vijayasree Vayalanellore , Barichello Tatiana 
Published: January 2023

Abstract
Evidence suggest that prenatal immune system disturbance contributes largely to the pathophysiology of neuropsychiatric disorders. A bacterial infection animal model widely accepted is administration of lipopolysaccharide (LPS). We investigated if maternal immune activation (MIA) could induce inflammatory alterations, oxidative damage and damage to blood brain barrier integrity (BBB) and placental barrier (PB) in fetal brain and pregnant Wistar rats. On gestational day 15, Wistar pregnant rats received LPS (0.25 mg/kg) to induce MIA. After 6, 12 and 24 h, fetus brain, placenta, and amniotic fluid were collected to evaluate early effects of LPS. Other group of rats from postnatal day (PND) 54 received injection of ketamine at the doses of 5, 15, and 25 mg/kg. On PND 60 rats were subjected to the spontaneous locomotor activity and pre-pulse inhibition test (PPI). MIA increased oxidative stress in the amniotic fluid and fetal brain. The BBB integrity in the hippocampus and cerebral cortex as well integrity of PB in the placenta and fetus brain were dysregulated after LPS induction. We also found elevated levels of cytokines such as interleukin (IL)-1β, IL-2, IL-4, IL-5, interferon (IFN)-γ, macrophage colony stimulating factor (M-CSF), IL-6, IL-10, IL-18, macrophage inflammatory protein (MIP)-3a, tumor necrosis factor (TNF)-α-, IL-7, MIP-1a and erythropoietin. Rats that receive MIA plus ketamine had locomotor activity impairment and a deficit in the PPI. In the present study, it was concluded that MIA increased the permeability of BBB and PB, the circulation of inflammatory mediators in fetal brains. This immune response in the prenatal period may be responsible for the behavioral changes in the adult life of offspring induced by MIA.
RNA editing is required for innate immune homeostasis through inhibiting cytosolic RNA receptor MDA-5 activated by cellular endogenous RNAs
Authors: Wang Qingde , Li Xiaoni , Qi Ruofan , Zhang Wenshuo , Zheng Pengyuan , Billiar Timothy R. 
Published: January 2023

Abstract
Cytosolic RNA receptors, RLRs, play an essential role in antiviral infections that specifically detect the invaded viral RNAs leading to IFN and ISG expression and antiviral reactions through RNA sensing signaling pathways. This viral RNA sensing mechanism tolerate cellular “self” RNA in normal cells. However, how RLRs distinguish viral and self RNA molecules remains ambiguous. Our recent study showed that insufficient RNA editing by editing enzyme ADAR1 enables the cellular endogenous RNAs to stimulate the RLRs and activate the innate immune signaling pathway that lead to type I IFN and ISGs expression. Specific deletion of ADAR1 in the hepatocytes in a mouse model causes profound inflammation in the liver. As a result, the mortality rate is dramatically increased in the young animals. However, deletion of MDA-5 in these ADAR1 KO mice completely rescues the mice, and all the inflammatory phenotype was reversed. This finding indicated that RNA editing is necessary for the innate immune homeostasis, and MDA-5 plays a critical role in sensing endogenous self RNAs. Studies on this endogenous RNA sensing signaling pathway, its regulation and potential involvement in autoimmune and inflammatory diseases is warranted.
Differential signaling through TLR7 or TLR8 determines the phenotype of human monocytes during RNA virus infection
Authors: Dominguez-Villar Margarita , de Marcken Marine , Dhaliwal Khushwant 
Published: January 2023

Abstract
Despite being a major cell population in blood, and one of the major cellular targets of many RNA virus infections in peripheral blood, the molecular consequences of interactions between human monocytes and RNA viruses, and the signaling pathways responsible for the activation of these cells during infection are not well understood. Toll-like receptors (TLR) are a major family of pattern recognition sensors that trigger specific activation pathways in cells from both the innate and adaptive arms of the immune system. There are at least 10 TLRs in humans, from which TLR7 and TLR8 recognize single-stranded RNA. Despite both recognizing the same general ligand, we and others have demonstrated different phenotypic outcomes on cells stimulated through either TLR7 or TLR8. Our laboratory has recently observed fundamental differences in the phenotype and function of monocytes stimulated via either TLR7 or TLR8, in the context of RNA virus infections, and specifically, in terms of type I IFN responses and effector cytokines they produce, as well as differences in cell surface markers. We have defined the molecular mediators that are responsible for these differences in phenotype, performing ex vivo experiments with human monocytes isolated from blood and we have shown the relevance of these data in common RNA virus infections, demonstrating that TLR7 and/or TLR8 stimulation by RNA viruses in human monocytes accounts for much of the phenotype the cells acquire upon virus interaction.
Stingray venom activates IL-33 producing cardiomyocytes, but not mast cell, to promote acute neutrophil-mediated injury
Authors: Silva Carla Lima , dos Santos Janaina Cardoso , Grund Lidiane Zito , Seibert Carla Simone , Marques Elineide Eugenio , Soares Anderson Brito , Quesniaux Valerie F. , Ryffel Bernhard , Lopes-Ferreira Monica 
Published: January 2023

Abstract
One of the hallmarks of acute inflammation is neutrophil infiltration of tissues. We investigated molecular mechanisms implicated in acute neutrophilic inflammation induced by the venom of a freshwater stingray (Potamotrygon cf. henlei) in mice. Ray venom induced early mobilization of neutrophil in the microvasculature of cremaster mice and infiltration of the peritoneal cavity 2 hours after injury, in a dose-response manner. IL-1b, IL-6, TNF-a, and KC were produced. The neutrophilic infiltration did not occur in mice with ST2 receptor and MyD88 adapters neutralized, or in those with PI3K and p38 MAPK signaling blocked. Drastic reduction of neutrophil infiltration to peritoneal cavities was observed in ST2−/−, TLR2/TLR4−/−, MyD88−/−, TRIF−/− and IL-17A−/− mice, and a partial reduction was observed in IL-18R−/− mice. Mast cell Kit W(sh)/W(sh)-, AHR-, NLRP3-, ICE-, IL-1b-, P2RX7-, CD39-, IL-17RA-, and TBX21 KO mice retain the ability to induce neutrophilia in peritoneal cavity after ray venom injection. IL-6 and TNF-a alone were insufficient for promote neutrophilia in the absence of ST2 signaling. Finally, abundant production of IL-33 by cardiomyocytes was observed. These results refine our understanding of the importance of the IL-33/ST2 axis and IL-33-producing cardiomyocytes in the early acute neutrophilia induced by freshwater stingray venoms.
C-reactive protein is an atheroprotective molecule
Authors: Pathak Asmita , Singh Sanjay K. , Agrawal Alok 
Published: January 2023

Abstract
The co-localization of plasma C-reactive protein (CRP) and atherogenic low-density lipoprotein (LDL) at the atherosclerotic lesions raises the possibility of a role of CRP in the disease process. In vitro, in its native pentameric structural form, CRP does not bind to oxidized LDL. However, in its non-native pentameric structural form, CRP is capable of binding to oxidized LDL. It has been proposed that CRP changes its structure at sites of inflammation to gain the oxidized LDL-binding activity. In vivo, native CRP is neither pro-atherosclerotic nor atheroprotective in animal models of atherosclerosis. We assumed that native CRP shows no effect because inflammatory microenvironment in the arterial wall in animal models of atherosclerosis is not appropriate and, therefore, the structure of CRP remains unchanged. Accordingly, we hypothesized that a CRP mutant, generated by site-directed mutagenesis, capable of binding to oxidized LDL without the requirement of any further structural change should show an effect on the disease. In the current study, we evaluated the impact of such a CRP mutant on the development of atherosclerosis employing LDL receptor knockout mouse model of atherosclerosis. We found that there was a two-week delay in the development of atherosclerotic lesions in the aortic root of mice treated with mutant CRP compared to mice that did not receive CRP. The development of atherosclerotic lesions in the whole aorta was also reduced; there was 30% reduction in the size of lesions in mice treated with mutant CRP. Overall, the data indicate that CRP is indeed an atheroprotective molecule.
Group 3 innate lymhoid cells are absent in DOCK8-defective HIES patients
Authors: Eken Ahmet , Okus Fatma Zehra , Patiroglu Turkan , Erdem Serife , Karakukcu Musa , Cansever Murat , Donmez-Altuntas Hamiyet , Canatan Halit , Oukka Mohamed , Topal Erdem , Kiyim Ayca , Karakoc Elif , Metin Ayse , Reisli Ismail , Uygun Vedat , Baris Safa , Keles Sevgi , Ozen Ahmet , Karasu Gulsun , Unal Ekrem 
Published: January 2023

Abstract
Innate lymphoid cell (ILC) subsets ILC1, ILC2 and ILC3 mirror image the functions helper CD4+ T cell subsets, Th1, Th2 and Th17, respectively; rely on similar transcription factors for their development, and produce similar effector cytokines. Thus, they have recently been shown to be crucial for protective immunity in mice. More importantly, in several chronic inflammatory diseases activated or increased frequency of ILCs have been reported in the circulation or affected tissues of patients. Our previous work with Dock8-defective Dock8pri/pri mice revealed that Dock8 is required for the development/function and survival of murine ILC3s. Thus Dock8pri/pri mice lacked ILC3s and was susceptible to Citrobacter rodentium infections. However to date, whether DOCK8 regulates ILC3 development or functions in humans has not been addressed. In the current study, using 11 DOCK8 mutant patients from across Turkey, we show, for the first time, that humans with DOKC8 deficiency lack peripheral ILC3s. Reduction in blood ILC3 could be verified by flow cytometry. Additionally, sorted total blood ILC population obtained from HIES patients with DOCK8 mutations shows diminished ILC3-specific Gm-csf, AhR and il23r gene expression in line with ILC3 loss. The defect is dramatic in ILC3s and less so in ILC2s in circulation. Flow cytometric examination of 11 patients revealed full restroration of ILC3s following hematopoietic stem cell transplantation. We also show that sorted peripheral ILC precursor population obtained from DOCK8 mutant patients has impaired capacity to proliferate in ILC1-ILC2-ILC3 polarizing conditions. This is the first report in the literature to show a selective deletion of ILC3s in HIES patients with DOCK8 mutations.
The Role of Glycogen Metabolism in Glycolytic Reprogramming Required for Dendritic Cell Immune Responses
Authors: Amiel Eyal , Thwe Phyu M. 
Published: January 2023

Abstract
Dendritic cells (DCs) are potent professional antigen presenting cells of the immune system and serve as a bridge between innate and adaptive immune responses. Activated DCs undergo a process of maturation defined by cytokine & chemokine secretion, co-stimulatory molecule expression, antigen processing and presentation, and the ability to activate T cells. DC maturation is coupled with an increase in energy demand, which is fulfilled by a TLR-driven burst in glycolytic metabolism. Up-regulation of glycolysis in activated DCs provides these cells with molecular building blocks and cellular energy required for DC effector function. Inhibition of glycolysis impairs both the survival and effector function of activated DCs. According to the prevailing model, TLR-driven glycolysis in DCs is sustained primarily by increases in extracellular glucose uptake. While non-immune cells, specifically hepatocytes and muscle cells, can store glucose in the form of glycogen as an intracellular energy reserve, the role of glycogen metabolism in supporting DC immune responses has not previously been described. Our data show that DCs possess intracellular glycogen stores and express enzymes essential for glycogen metabolism. Furthermore, inhibition of glycogen metabolism significantly impairs key activation parameters and effector functions of these cells. We further report that DCs exhibit functional compartmentalization of glucose- and glycogen-derived carbons, where these substrates preferentially contribute to distinct metabolic pathways. This work provides novel insights into nutrient homeostasis in DCs, demonstrating that differential utilization of glycogen and glucose metabolism regulates their optimal immune function.
Ly49H-dependent IFNγ protein production by NK cells requires cytokine signaling that induces IFNγ mRNA transcription.
Authors: Piersma Sytse J. , Pak-Wittel Melissa A. , Yokoyama Wayne M. 
Published: January 2023

Abstract
Natural killer (NK) cells recognize and attack target cells, including cancer and pathogen-infected cells, through a combination of activating and inhibitory signals that follow receptor–ligand interactions. NK cells play a critical role in controlling murine cytomegalovirus (MCMV) infection in C57BL/6 mice, which is dependent on interactions between the Ly49H NK cell activation receptor and its MCMV-encoded ligand m157. This interaction is not sufficient to activate NK cells and control MCMV infection. We previously reported that IL-12 and type I IFN produced early during MCMV infection induce granzyme B and perforin in NK cells, which allows them to efficiently kill virus-infected cells upon recognition of m157 through Ly49H. We now show that m157-dependent IFNγ production also requires cytokine signaling. Using quantitative PCR, the PrimeFlow RNA assay, and chemical inhibitors we found that cytokine signaling was required for IFNγ mRNA transcription, whereas signaling through Ly49H resulted in translation of IFNγ mRNA into protein. Furthermore, initial exposure of NK cell to cytokines enables them to produce IFNγ protein later in the absence of cytokines. We propose the following model: MCMV infection induces cytokine production including IFN-I and IL-12. These cytokines induce the production of cytolytic proteins and IFNγ mRNA. As a result, the NK cells are poised and ready for attack. Virus-infected cells are subsequently recognized by NK cells trough activating ligands such as m157. This activation signaling results in IFNγ production and the release of lytic granules, which results in elimination of the virus infected cells. Together these two pathways enable NK cells to swiftly attack and eradicate virus infections.
Role of telomerase expression in T cell differentiation and proliferation
Authors: Patrick Michael S. , Cheng Nai-Lin , Kim Jaekwan , Dong Fangyuan , An Jie , Weng Nan-Ping 
Published: January 2023

Abstract
To understand the role of telomerase in T cell differentiation and function, we analyzed human telomerase reverse transcriptase (hTERT) mRNA expression and telomerase activity in six T cell subsets from 85 healthy human donors (aged 17–82 years old). We found that levels of hTERT mRNA and telomerase activity were higher in CD4 T cell subsets as compared to corresponding CD8 T cell subsets, and decreased from naïve (TN) to memory (central, TCM and effector, TEM). In all six subsets, in vitro activation with anti-CD3/CD28 antibody-coated beads led to increased amounts of hTERT mRNA (from 12 to 105-fold) as well as telomerase enzymatic activity, while the differences in hTERT amounts and telomerase activity among subsets remained in the same order observed prior to stimulation. We next analyzed the impact of different amounts of hTERT and induced telomerase activity on proliferation and lifespan of these subsets in vitro. We found that proliferation and survival were most robust in CD4 TN and poorest in CD8 TEM, correlating with their respective levels of hTERT mRNA and telomerase activity both pre-activation and over the course of a 15-day culture following a single stimulation. These findings suggest that greater hTERT expression and telomerase activity promote T cell viability and proliferative potential, and that differentiation is associated with loss of proliferative potential in T cells. We are currently analyzing whether inhibition of hTERT and telomerase activity will lead to reduced proliferation and viability in TN cells.
This research was supported entirely by the Intramural Research Program of the NIH, National Institute on Aging.