bluuebunny/crossref_metadata_2025_split huggingface dataset:
Macrophage plasticity and activation states drive differential expression of murine macrophage galactose-type lectin (MGL) orthologs
Authors: Paez Reina N , Browning Sarah , Files Megan A , Varghese Alyssa A , Huante Matthew B , Strong Emily , Lee Sunhee , Endsley Mark A , Endsley Janice J 
Published: August 2023

Abstract
Macrophage plasticity and functional activation are critical to the broad array of macrophage-mediated functions that help determine disease outcomes. Macrophage galactose-type lectin (MGL), a type-2 C-type lectin receptor (CLR) primarily expressed on myeloid cells, has been shown to play an anti-inflammatory role in the innate response to some infectious agents. Our lab recently demonstrated that MGL-1 contributes an antimicrobial and anti-inflammatory role in a murine model of experimental tuberculosis. We further observe that MGL-1 and MGL-2 expression by RAW 264.7 macrophages are similarly activated by whole mycobacteria, but differentially regulated by signaling through toll-like receptors (TLR) as well as cytokine and glucocorticoid receptors. We find that synthetic agonists and microbial ligands of TLR2, TLR4, and TLR7 positively regulate MGL-2 expression, suggesting responsiveness to a broad repertoire of antigens compared to MGL-1. Additionally, activation of MGL-2 expression differs between ligands of multiple bacterial species, both pathogenic and non-pathogenic. Pathway inhibition experiments show that MGL-2 upregulation by microbial stimuli occurs through MyD88-dependent Toll-like receptor (TLR) signaling. Although both MGL-1 and MGL-2 are classically associated with alternatively activated macrophages, we show that MGL-2 expression is associated with both M1 and M2 polarized macrophages, while MGL-1 expression is restricted to M2. These results demonstrate a strong relationship between macrophage activation states and the differential activation of MGL signaling and indicate non-redundant roles of MGL-1 and MGL-2 in innate immunity of murine models of disease.
Supported by grants from NIH, NIAID (R61 AI138328, R33 AI138328)
Secondary cues alter ISG protein repertoires to unlock antiviral subprograms triggered by interferons
Authors: Hancks Dustin , Chang Tyron 
Published: August 2023

Abstract
Immune defenses are triggered by cytokines like interferon-α (IFN-α) and IFN-γ in diverse cells. Both IFNs induce a class of factors termed IFN-stimulated genes (ISGs). Many ISG proteins have antiviral functions. There is a growing appreciation that ISG repertoires are cell-type specific and that this bias impacts infection outcomes. However, the cues and mechanisms driving these variable responses are ill-defined. Given key roles for immunometabolism in immune cell activation, we hypothesized that local cues like metabolites shape tissue specific ISG defenses. To model the impact of metabolic cues on host defenses in non-immune cells, we infected human cells with two unrelated viruses: vaccinia virus, the prototypical poxvirus, and herpes simplex virus-1 (HSV-1). Unexpectedly, we find that cells favoring glycolysis, but not OXPHOS, catalyze potent IFN-γ restriction of vaccinia and HSV-1 that attenuate viral-gene and protein expression as well as infectious virus production. This response is specific as modulating IFN-α signaling by supplementation with either glycolysis or OXPHOS cues has no effect on viral replication. This interferon-induced subprogram is independent of differences in canonical IFN-γ signal transduction, occurs at low levels of IFN-γ, and induced by physiological levels of glucose. RNA-seq reveals no marked changes in the “usual suspects” (e.g. ISGs). Instead, we find secondary cues regulate ISG protein levels. Deletion of a single ISG protein, which is differentially stabilized by secondary cues, is sufficient to rescue vaccinia replication. These data indicate that secondary cues can alter infection outcomes in the same cell-type by unlocking subprograms embedded in interferon catalyzed responses.
DNA-PKcs regulates the cGAS-STING signaling axis during infection with intracellular bacterium Listeria monocytogenes
Authors: Morales Abigail , Dominguez Zuley A , Rodriguez Ricardo A , Morales Rosalie , Dilgen Teague C , Flores Melissa 
Published: August 2023

Abstract
The cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) (cGAMP) synthase (cGAS) and the ER-localized stimulator of interferon genes (STING) are critical components of innate immune signaling. Though the intracellular signaling pathway was first characterized as a critical host defense against viral infection, recent evidence has suggested that the pathway is also activated during infection with some types of bacteria, though the importance is complicated by the differential impact of type I interferon on the progression of distinct bacterial infections: in response to some species of bacteria, type I interferon production is detrimental, while it has a protective function during infection with others. DNA-PKcs is a critical kinase in the repair of and response to DNA double-strand breaks (DNA DSBs) in mammalian cells. Several recent reports have indicated that it also regulates the cGAS-STING signaling axis in virally-infected cells, suggesting that it may have previously unappreciated roles in regulating cytoplasmic signaling. Notably, we find that DNA-PKcs is required for optimal transcriptional induction of IFN-β and a number of other well-characterized interferon-stimulated genes (ISGs) in macrophages during infection with L. monocytogenes. Additionally, we find that phosphorylation of kinase TBK1 and activation of IRF3 are significantly diminished in Scid (DNA-PKcs−/−) macrophages relative to wildtype controls during infection with L. monocytogenes. As type I interferon is detrimental to effective Listeria clearance in murine infection models, these findings may provide an additional rational explanation as to why Scid mice clear Listeria more efficiently than their wildtype counterparts.
DNA-PKcs Regulates Transcriptional Networks in Activated Macrophages
Authors: Morales Abigail , Morales Rosalie , Dominguez Zuley A , Dilgen Teague C , Flores Melissa , Rodriguez Ricardo A , Winkler Michelle 
Published: August 2023

Abstract
The maintenance of genome integrity is critical for organismal survival; however, eukaryotic cells perpetually sustain various types of DNA damage. DNA double-strand breaks (DSBs) are the most insidious of these lesions, as they can lead to chromosomal translocations if they persist unrepaired. A substantial body of work has established, however, that DNA DSBs are also critical signaling intermediates that impact a variety of cell-type-specific processes. Recently published work established that upon infection with the intracellular bacterium Listeria monocytogenes, activated macrophages sustain damage in their genomic DNA. This initiates a DNA damage response (DDR) that depends on both ATM and DNA-PKcs kinases. Though ATM has been implicated in the regulation of transcriptional networks downstream of DNA damage in many different cellular contexts, few studies have directly addressed a potential role for DNA-PKcs in regulating gene expression. Thus, we asked whether DNA-PKcs kinase activity is important for regulating transcriptional responses in macrophages in settings of inflammation. In particular, we asked whether DNA-PKcs kinase activity is required for optimal activation of NF-κB. Our preliminary data suggest reduced translocation of NF-κB to the nucleus in the absence of DNA-PKcs kinase activity with a corresponding diminished expression in key NF-κB-regulated pro-inflammatory genes. As NF-κB is also activated in response to DNA damage, we suggest that in macrophages, pathogen-induced TLR signaling and DNA damage signaling synergize to drive NF-κB-mediated transcriptional responses, thereby regulating the cell’s function in the innate immune response to infection.
A Role for Stimulator of Interferon Genes in Toll-Like Receptor 8 Signaling
Authors: García-Martínez Karla Yvonne , Chen Jingyi , Leifer Cynthia A 
Published: August 2023

Abstract
The innate immune system is equipped with multiple receptors to detect microbial nucleic acids and induce type I interferon (IFN) to restrict viral replication. When dysregulated these receptor pathways induce inflammation in response to host nucleic acids and promote development and persistence of autoimmune diseases like Systemic Lupus Erythematosus (SLE). IFN production is regulated by the Interferon Regulatory Factor (IRF) transcription factor family downstream of several innate immune receptors such as Toll-like receptors (TLRs) and Stimulator of Interferon Genes (STING). Both TLRs and STING activate TANK-binding kinase I (TBKI) and IkappaB kinase-ɛ (IKKɛ), which phosphorylate and activate IRFs. The pathway by which TLRs and STING activate IFN response are considered independent.
Here we show that STING plays a previously undescribed role in human TLR8 signaling. Stimulation with TL8-506 (TL8), TLR8 ligand, induced IFN secretion in primary human monocytes and THP-1 cells and was reduced by inhibition of STING. Using THP-1 cells encoding reporters for IRF and NFkB activity, two major signaling pathways downstream of TLRs, we found that TL8 stimulation increased IRF and NFkB activity. When STING was inhibited, IRF, but not NFkB, activity was reduced. TL8-induced IRF activity was dependent on IKKɛ, not TBKI. Bulk RNA transcriptomic analysis show that TLR8 induces gene signatures associated with SLE which are downregulated by inhibition of STING. These data demonstrate that STING is required for full TLR8-IRF signaling.
These studies build a new framework for crosstalk between cytosolic and endosomal innate immune receptors, which could be used to treat IFN driven autoimmune diseases.
Rosiglitazone improves alveolar macrophage mitochondrial derangements induced by chronic alcohol consumption and infection with Klebsiella pneumoniae
Authors: Yeligar Samantha M 
Published: August 2023

Abstract

Objective
People with alcohol use disorders (AUDs) are 2–4 times more likely to develop lung infections, sepsis, and acute respiratory distress syndrome. Chronic alcohol misuse blunts alveolar macrophage (AM) immunity, in part, due to decreased peroxisome proliferator-activated receptor (PPAR)γ which increases cellular oxidative stress. Mitochondrial (MT) oxidative stress may impair AM phagocytosis; thus, we posited that chronic alcohol plus acute Klebsiella pneumoniae (Kp) dysregulates MT and treatment with rosiglitazone (RSG), a PPARγ ligand, reverses these effects to improve AM phagocytosis of Kp.

Methods
AMs were isolated from 8–10-month-old male C57BL/6J mice fed ± ethanol (EtOH, 20% wt/vol in drinking water) for 12 wks ± intratracheal 2 × 104 CFU of Kp in saline vehicle for the last 24 h of water or EtOH ± RSG (10 mg/kg/d in methylcellulose vehicle) via oral gavage 4 h prior to sacrifice (n=5/group). MT O2•− and H2O2 were assessed by fluorescence microscopy. mRNA and protein levels of PPARγ and MT dynamics mediators, transcription factor A mitochondrial (TFAM), dynamin-related protein 1 (DRP1), and mitofusin-2 (MFN2) were measured by qRT-PCR and cytoimmunostaining. Kp clearance by AMs was determined by plating lysed AMs on MacConkey agar.

Results
EtOH feeding to mice increased MT oxidative stress; decreased PPARγ, TFAM, and MFN2 levels; and increased DRP1 levels in AMs. Kp exacerbated EtOH’s MT derangements by ~51–63% and impaired Kp clearance in AMs by 57.3 ± 11.2%. RSG treatment in vivo reversed EtOH- and Kp-induced MT derangements in AMs and restored AM clearance of Kp.

Conclusion
RSG may provide a novel therapeutic strategy for mitigating AM MT derangements and immune dysfunction in AUD individuals infected with Kp.

NIAAA R01-AA026086
LRRK2 G2019S promotes the development of colon cancer
Authors: Wang Yuhang , Gao Joyce , Xiong Yiqin , Kang Zizhen 
Published: August 2023

Abstract
LRRK2-G2019S is the most prevalent variant associated with Parkinson’s disease (PD), found in 1–3% of sporadic and 4–8% of familial PD cases. Intriguingly, emerging clinical studies have suggested that LRRK2 G2019S mutation carriers have an overall increased risk of cancer, including colon cancer. However, it remains to be experimentally tested whether and how this mutant directly affects cancer development. Here, by using the AOM/DSS-induced colitis-associated colon cancer model combined with LRRK2 G2019S knock-in (KI) mice, we found that LRRK2 G2019S promoted the pathogenesis of colon cancer, characterized by increased tumor loads and tumor-promoting cytokines compared with those in wild-type controls. Immunoblotting and immunofluorescence indicated that expression levels of Cyclin-D1, Ki-67, β-catenin and p-STAT3 were dramatically increased in the colon of KI mice, and these proteins are well known to promote the pathogenesis of colon cancer. As inflammation is a major factor contributing to the progression of colon cancer, we thus further tested if LRRK2 G2019S promotes colitis in the initiation of the disease. Indeed, we found KI mice were more susceptible to colitis induction, evident by rapid body weight loss and elevated clinical histology score. Mechanistically, LRRK2 G2019S promoted gut inflammasome activation and enhanced the activation of the NF-ƙB and MAPK pathways upon Dectin-1 stimulation. Together, our data for the first time provide direct evidence that gain-of-function of kinase activity in LRRK2 promotes colon cancer development, implicating LRRK2 as a potential target in colon cancer, especially in those LRRK2 G2019S carrier patients and cancer with hyper LRRK2 expression.
Startup fund from the Department of pathology at the University of IOWA, NIH R21AG076895, R01 NS104164
An animal model for autoinflammation with infantile enterocolitis
Authors: Wang Yuhang , Gao Joyce Ziyi , Xiong Yiqin , Kang Zizhen 
Published: August 2023

Abstract
Autoinflammation with infantile enterocolitis (AIFEC) is one of the emerging life-threatening diseases caused by inborn errors of NLRC4 inflammasome. The pathogenicity of AIFEC remains largely unknown and standard therapy is lacking due to rare patient cases and high mortality. To fill the gap, it is urgent to set up an effective animal model for this rare disease. We have recently generated a loxP-flanked NLRC4 V341A conditional knock-in (KI) mouse line where human V341A (GTG to GCG) mutation was introduced into the corresponding mouse Nlrc4 coding sequence. When this line is bred with distinct Cre transgenic mice, the KI allele can be expressed globally or cell-type specifically. We bred the KI line with E2a-Cre transgenic mice and got globally expressed V341A KI mice, and the KI mice developed autoinflammation with infantile enterocolitis that recapitulated the symptoms of AIFEC patients. In brief, immunoblot demonstrated KI mice had hyperactivation of NLRC4 inflammasome in gut epithelium and macrophages. The neonatal KI pups had significant intestinal inflammation, accompanied with diarrheal, impaired barrier integrity and dysbiosis. Furthermore, the KI pups had dramatically elevated IL-1β, IL-6 and IL-18 in sera and supernatant of colon explant cultures. Meanwhile, the KI pups exhibited cytopenia, hemophagocytosis, and macrophage hyperactivation. Biochemical assays suggested multi-organ damage in KI mice, including the liver, kidney, and heart, accompanied with hyper serum ferritin level. Without any intervention, the pups died before day 10 postnatally. Taken together, we have established a novel animal model for AIFEC, thus providing a platform to elucidate the pathogenesis of AIFEC and test therapeutic strategies.
Startup fund from the Department of pathology at the University of IOWA, NIH R21AG076895, R01 NS104164
Regulation of Inflammatory NF-kB Target Gene Activation by Jak-STAT Signaling
Authors: Mishra Bikash , Ivashkiv Lionel 
Published: August 2023

Abstract
Macrophages are key innate immune cells that are important for host defense but their aberrant activation by IFN-γ has been implicated in the pathogenesis of chronic inflammatory diseases such as rheumatoid arthritis (RA). IFN-γ, signals through protein tyrosine kinases JAK1/2 to activate transcription factor STAT1 and drive the expression of interferon-stimulated genes (ISGs). IFN-γ primed macrophage induced rapid and enhanced expression of both canonical and non-canonical ISGs when challenged with toll-like receptor 4 (TLR4) ligand (LPS). However, IFN-γ alone was not able to elicit the expression of the noncanonical ISGs that are considered proinflammatory NF-kB target genes. Based on this observation, we hypothesized that any inflammatory stimuli can synergistically activate NF-kB target genes in IFN-γ primed macrophages. Like LPS, TLR2 ligand (PAM3) synergistically induced NF-kB target genes expression but inflammatory cytokine TNF-α failed to do so in IFN-γ primed macrophages. The primed macrophage is similar to RA macrophage where JAK signaling is ongoing and STAT1 is activated in tandem with inflammatory NF-kB signaling. Our model suggests that targeted inhibition of JAK1/2 could be a potential strategy to inhibit the production of inflammatory cytokines by these macrophages. To address this, we used an FDA-approved JAK1/2 inhibitor for RA. JAK inhibition effectively suppressed the expression of a subset of IFN-γ-induced genes, whereas LPS induced genes remained partially resistant to inhibition. This suggests that IFN-γ priming induces long term epigenetic memory which persist even after inhibition of IFN-γ signaling.
GATA2-TGF-b1 axis regulates human NK cell development
Authors: Wang Dandan , Myers Kasiani C , Bresnick Emery H , Verbsky James , Thakar Monica , Malarkannan Subramaniam 
Published: August 2023

Abstract
Natural killer (NK) cells, a subset of innate lymphocytes, produce proinflammatory cytokines and mediate anti-tumor cytotoxicity. GATA2 is a master transcription factor that is essential for the development of erythroid, myeloid, B, and NK cells. Patients with GATA2 haploinsufficiency lack CD56bright NK cells (immature), with or without a reduction in the number of CD56dim NK cells (mature). However, how GATA2 functions to establish and maintain genetic networks during the development and function of human NK cells is unknown. Here, we identify a novel GATA2-TGF-b1 axis that regulates NK cell development. We performed single-cell RNA-seq with NK cells from three GATA2T354M patients and found significantly reduced expression of immediate early genes, which indicated that they are functionally-defective. We discovered a reduction in TGF-b1 transcripts and defective expression of TGF-b1 target genes in NK cells from GATA2T354M patients. Using a reporter assay, we determined that GATA2 occupy to TGFB1 promoter, and this association was decreased in GATA2T354M NK cells. ATAC-seq of GATA2T354M NK cells indicates significantly altered chromatin accessibility. GATA2 CUT&Tag-seq validated that it occupies the TGFB1 promoter region, which correlated with activation of the TGFB1 locus in human NK cells. In summary, we define a mechanism by which GATA2 controls TGF-b1 production in human NK cells and thereby regulating their development. These findings provide vital clues for developmental and functional defects of NK cells in GATA2T354M patients.
ASH Graduate Hematology Award (D.W.); GIRT Award from MCW-C4I (D.W.); NIH R01 AI102893 and NCI R01 CA179363 (S.M.); HRHM Program of MACC Fund (S.M.), Nicholas Family Foundation (S.M.); Gardetto Family (S.M.); MCW-Cancer Center-Large Seed Grant (S.M. and M.S.T.); MACC Fund (S.M.); Ann’s Hope Mela- noma Foundation (S.M.); and Advancing Healthier Wisconsin (S.M.)
Pellino1 promotes immunity in the skin during infection with herpes simplex virus (HSV)
Authors: Jensen Liselotte E , Cai Kathy Q 
Published: August 2023

Abstract
Pellino1 (Peli1) is a ubiquitin ligase involved in IL-1 and Toll-like receptor signaling; however, the role of Pellino1 in skin immunity against HSV is unknown. Here, using the mouse flank HSV-1 skin infection model, we demonstrate that Pellino1 has several critical functions during active viral replication. Analyses of single cell genome wide gene expression data (RNA-seq) revealed expression of Peli1 mRNA throughout the skin including in epithelial, fibroblast and immune cells. Both histology and RNA-seq demonstrated normal development of the epidermis and dermis in Pellino1 knockout (KO) mice. The median survival of Pellino1 KO mice was 12 days compared to greater than 16 days in wild type mice. Pellino1 deficient mice also developed larger zosteriform skin lesions along affected dermatomes. During infection HSV-1 did not replicate in the dermis. Immunohistochemistry revealed delayed recruitment of myeloid and T cells to early infected epidermis in Pellino1 deficient mice. This correlated with lower expression of the inflammatory IL-1 and IL-36 cytokines and the poorly understood chemokine Grp15l. In Pellino1 KO mice, the virus spread extensively through the epidermis, follicular infundibulum into sebaceous glands where sebocytes were found positive for the virus. Histological analyses for HSV-1 and neuronal axons revealed nerve endings throughout the epidermis and axons wrapped around the hair follicles. Dissemination of HSV-1 into the sebaceous glands did not appear to involve a shift in how the virus migrated through these nerve axons as early lesions were only found in the interfollicular epidermis. In conclusion, Pellino1 plays important roles in restricting viral dissemination.
Supported by grant from NIH (R01 AI125111)
Butyrate modulation of inflammatory responses to lipopolysaccharide in porcine monocytes
Authors: Becker Sage R , Loving Crystal 
Published: August 2023

Abstract
Butyrate is a short-chain fatty acid produced through microbial fermentation of dietary fibers in the lower intestinal tract. Butyrate production is associated with enhanced mucosal immunity, increased barrier integrity, and modulation of immune cells. Butyrate driven immunomodulation occurs through a variety of mechanisms, including histone deacetylase (HDAC) inhibitor activity and inhibition of NF-kB signaling. Butyrate impacts lipopolysaccharide (LPS)-induced cytokine production and gene expression by porcine monocytes in a dose-dependent manner. To investigate the mechanisms driving butyrate modulation of porcine monocyte responses to LPS, peripheral monocytes were cultured with LPS +/− butyrate. After incubation, cells were processed for collection of cytoplasmic and nuclear fractions. HDAC activity in nuclear fractions and NF-kB signaling state (IkBa, p-IkBa, p65, p-p65) in cytoplasmic fractions was assessed. Butyrate did not inhibit HDAC activity after 1 hour (h) but at 24 h, butyrate exposure resulted in inhibition of HDAC activity regardless of LPS exposure, in a dose-dependent manner. Neither the low nor high concentration of butyrate altered IkBa, p-IkBa, p65, or p-p65 status, though p-IkBa and p-p65 status was dependent on LPS exposure. Overall, the impact of butyrate on HDAC activity was time and dose-dependent, and butyrate had little to no effect on NF-kB signaling induced by LPS exposure. Continued investigation is warranted to understand the potential mechanisms driving butyrate modulation of monocyte responses to LPS to enhance abilities to modulate immune status.
USDA-ARS CRIS 005