bluuebunny/crossref_metadata_2025_split huggingface dataset:
PAK6- A novel kinase in the maintenance of T cell anergy
Authors: Torres Annabel , Tang Alice , Vyas Radha , Su Chen , Johnson Matthew , Wells Andrew 
Published: January 2023

Abstract
Costimulation of T cells through the antigen receptor and the CD28 receptor engages an ‘effector’ gene expression program and a burst of proliferation. Conversely, antigen receptor engagement in the absence of costimulation induces a hypo-responsive state, referred to as anergy, that is associated with active silencing of effector genes. Our ATAC-seq analysis of CD28-dependent chromatin remodeling in anergic and effector CD4+ T cells has identified p21-activated kinase 6 (PAK6) as a gene that becomes accessible in anergic but not effector T cells. PAK6 is a member of a family of kinases comprised of two groups, group I (PAK1-3) and group II (PAK4-6). PAK6 and other group II members lack a regulatory domain that is present in group I, causing it to function differently. Previous studies by others have shown that PAK2, a group I PAK, was a positive regulator of CD3/CD28 T cell activation. Here, we demonstrate that PAK6 has the opposite role in T cell activation as PAK2, where it inhibits IL2 secretion, a hallmark characteristic of T cell anergy. Knockdown of PAK6 in anergic and effector T cells, using a CRISPR/CAS9 approach, resulted in a 50% increase in IL2 secretion, and 5-fold more cell expansion in anergic cultures upon restimulation with CD3/CD28. This suggests that PAK6 may be involved in a signaling pathway which inhibits cellular proliferation or promotes apoptosis. The highest level of PAK6 expression was observed upon re-stimulation of anergic cells, suggesting that PAK6 may be involved in the maintenance of T cell anergy. This study identifies a novel role for PAK6 in T cell anergy and a potential new target for regulating T cell function in autoimmune diseases.
JNK inhibition by Transforming Growth Factor-β Type I Receptor (TGF-β RI) in T Cells
Authors: Jacks Ramiah , Iwashima Makio 
Published: January 2023

Abstract
T cells are a critical component of the adaptive immune system and their responses in pathogenic and steady state conditions require strict regulation. One mechanism involved in this regulation is Transforming Growth Factor-β (TGF-β) signaling. TGF-β acts on activated T cells to facilitate the differentiation of some T helper subsets or the suppression of activation and proliferation. Our recent data demonstrate that TGF-β plays a role in the survival of Tregs against continuous antigen exposure. TGF-β signaling is propagated by initially binding to the TGF-β type II receptor, which activates TGF-β type I receptor that phosphorylates the transcription factors SMAD2/SMAD3. This induces nuclear localization of SMAD complexes and transcription of target genes. To understand how TGF-β signaling is coupled to T cell specific events, we generated TGF-βRI deficient Jurkat T cells using the CRISPR/cas9 system. Unexpectedly, TGF-βRI deficient Jurkat demonstrate a hyperactivation phenotype featuring a highly elevated expression of surface antigens (CD69, CD25), and secreted significantly increased levels of IL-2 and TNF. Analysis of signaling events in TGF-βRI deficient Jurkat demonstrate constitutive activity of activator protein-1 (AP-1), constitutive activation of jun-amino-terminal kinase (JNK) and increased expression of JUN. Similarly, in primary T cells transient expression of a dominant negative mutant of TGF-βRI elevated AP-1 activity while a constitutively active mutant reduced this activity. Together, these data show that TGF-βRI signaling plays a novel role in controlling the activity of JNK and AP-1 in T cells. This signaling pathway may play a critical role in TGF-β mediated differentiation and/or survival of T cells.
Genome-wide analysis identifies pairs of cis-acting lncRNAs and protein-coding genes involved in innate immunity
Authors: Zhang Qiong , Chao Ti-chun , Patil Veena S. , Qin Yue , Tiwari Shashi Kant , Chiou Joshua , Dobin Alexander , Tsai Chih-Ming , Li Zhonghan , Dang Jason , Gupta Shagun , Urdahl Kevin B , Nizet Victor , Gingeras Thomas R. , Gaulton Kyle J , Rana Tariq M. 
Published: January 2023

Abstract
Long noncoding RNAs (lncRNAs) can regulate target gene expression by acting in cis (locally) or in trans (non-locally). Here, we performed genome-wide expression analysis of Toll-like receptor (TLR)-stimulated human macrophages to identify pairs of cis-acting lncRNAs and protein-coding genes involved in innate immunity. A total of 229 gene pairs were identified, many of which were commonly regulated by signaling through multiple TLRs and were involved in the cytokine responses to infection by group B Streptococcus. We focused on elucidating the function of one lncRNA, named lnc-MARCKS or ROCKI (Regulator of Cytokines and Inflammation), which was induced by multiple TLR stimuli and acted as a master regulator of inflammatory responses. ROCKI interacted with APEX1 (apurinic/apyrimidinic endodeoxyribonuclease 1) to form a ribonucleoprotein complex at the MARCKS promoter. In turn, ROCKI–APEX1 recruited the histone deacetylase HDAC1, which removed the H3K27ac modification from the promoter, thus reducing MARCKS transcription and subsequent Ca2+ signaling and inflammatory gene expression. Finally, genetic variants affecting ROCKI expression were linked to a reduced risk of certain inflammatory and infectious disease in humans, including inflammatory bowel disease and tuberculosis. Collectively, these data highlight the importance of cis-acting lncRNAs in TLR signaling, innate immunity, and pathophysiological inflammation.
Identification and characterization of fish-specific IL-23 isoforms and their roles in fish immunity
Authors: Qin Lei , Zhang Shengnan , Zhou Hong 
Published: January 2023

Abstract
IL-23 is well known as a regulator in inflammation response and host defense by promoting the development of Th17 cells. In fish, three genes coding p40 (p40a, p40b and p40c) are originated from fish-specific genome duplication, thereby generating three IL-23 isoforms (IL-23a, IL-23b and IL-23c). Considering the functional diversity of fish IL-23, it is possible that the fish-specific immunoregulatory mechanism of IL-23 may have existed. In this study, we isolated and identified the subunits of IL-23 from grass carp. In response to bacterial infection in vivo, the mRNA levels of gcp19 were upregulated in headkidney, liver and gill, while those of p40a were significantly induced in the selected tissues except liver. Surprisingly, the levels of gcp40b and gcp40c did not show marked change in tissues. What’s more, LPS, PHA stimulation in vitro, the mRNA levels of these subunits also showed a significant increase in periphery blood lymphocytes. Meanwhile, the result of Co-IP showed p19 could couple with each of the three p40 to form IL-23 and be secreted into the supernatants, verifying the existence of three IL-23 isoforms in grass carp. In order to assess their functions in fish immunity, we constructed plasmids containing gcp19 CDS combined with gcp40a/b/c CDS by a linker sequence and over-expressed them in CHO cells. The positive stable transfectants were selected, and the recombinant proteins were produced and purified from the culture medium. At present, we have gained these grass carp IL-23 proteins. Their bioactivities were investigated in grass carp headkidney cells, showing same or different functions in the immune regulation of grass carp. Finally, our work has provided a basis for elucidating the roles of IL-23 isoforms in fish immunity.
Cannabinoid receptor 1 blockade attenuates obesity and adipose tissue type 1 inflammation through miR-30e-5p regulation of Delta-like-4
Authors: Miranda Kathryn , Mehrpouya-Bahrami Pegah , Nagarkatti Prakash S , Nagarkatti Mitzi 
Published: January 2023

Abstract
Obesity is characterized by chronic low-grade inflammation that contributes to development of cardiometabolic disorders. Cannabinoid receptor 1 (CB1) antagonists attenuate diet-induced obesity (DIO) and related inflammation, although the precise anti-inflammatory mechanisms involved have not been fully explored. In the current study we used a mouse model of DIO intervention to determine the microRNA (miR)-mediated anti-obesity and anti-inflammatory effects of the CB1 antagonist AM251. DIO mice that were fed high-fat diet (HFD) for 12 weeks were treated with AM251 (10mg/kg) for an additional four weeks. HFD+AM251 mice experienced rapid and prolonged weight loss and reduced inflammatory M1 adipose tissue macrophage (ATM) infiltration. To investigate miRNA-mediated regulation of ATMs, F4/80+ cells from stromal vascular fractions of epididymal fat were subjected to miR microarray analysis. Several miRs were differentially expressed in AM251-treated mice that were independent of calorie restriction. Prominently, miR-30e-5p was upregulated in ATMs from HFD+AM251 mice while the miR-30e-5p targeting DLL4 was downregulated. Consistent with a decrease in DLL4-Notch signaling, fat storage and pro-inflammatory cytokine/chemokine expression was reduced following AM251 treatment. Furthermore, we demonstrate that AM251-treated macrophages can suppress DLL4-mediated Th1 polarization in CD4+ T cells. Together these data indicate that weight-loss due to AM251 treatment mediates anti-inflammatory effects and amelioration of obesity through miR-30e-5p regulation of DLL4-Notch signaling-induced type 1 inflammation. These data support therapeutic potential of miR-30 in the treatment of cardiometabolic disorders.
A host-associated antimicrobial agent exhibits both immunoprotective and immunotherapeutic effects
Authors: Dawes Maisie Ernestene , Xie Chen , Chitko-McKown Carol G. 
Published: January 2023

Abstract
Inflammation is a critical component of both the defense against injury and the pathophysiology of disease. Despite advances in anti-inflammatory therapy and critical care, Gram-negative infections remain a significant cause of mortality in multiple species, and are a primary cause of diarrhea, endotoxemia, sepsis and ultimately death in neonatal calves. Given the impact of host response to disease on the health of humans and food animals alike, we examined the effects of supplemental bovine lactoferrin (bLF) and its functional peptide lactoferricin B (LFcin B), on the dysregulated immune response to lipopolysaccharide (LPS). We hypothesized that these antibiotic agents have therapeutic value as low-risk anti-inflammatory agents, a characteristic that is attributable to their regulatory role on intracellular signaling in innate cells.
Although present in colostrum, saliva and mucosal secretions, bLF is notably at its highest concentration in the secondary granules of neutrophils. In the host, the 25 AA peptide LFcin B is released following the activity of proteolytic enzymes in the stomach as well as at sites of infection.
Viable peripheral monocytes and neutrophils were isolated from calves and stimulated in vitro with LPS, in the presence or absence of bLF/LFcin B. Samples were analyzed using qRT-PCR, Meso Scale Discovery Electrochemiluminescence, immunoprecipitation and western immunoblot detection. The effects of bLF and LFcin B on pro-inflammatory cytokine and inflammatory mediator production will be expressed as the mean ± SD and will be considered significant at P&lt;0.05. Although each exerted differential effects, the p38 MAP kinase pathway appears to be a likely target.
Bisphenol-A leads to overexpression of estrogen receptors and diminishes protective chemokine responses in a zebrafish model
Authors: Walser-Kuntz Debby , Tieu Tam , Scott Tamara , Khan Pavana 
Published: January 2023

Abstract
Bisphenol-A (BPA), widely used in thermal receipts and food containers, binds to both estrogen receptor (ER) α and β, disrupting estrogen activity and presenting an environmental challenge to the immune system of humans and wildlife. We are exploring the impact of in vivo BPA exposure on zebrafish innate immunity. Zebrafish exposed to 100 ng/ml BPA immediately following fertilization (day 0) show a two to fourfold reduction in gene expression of CXCL8, the chemokine that recruits neutrophils to the site of infection on days 3 through 6 post fertilization (dpf). To determine if reduced chemokine expression has a functional outcome, we studied MPO::GFP zebrafish expressing GFP under a neutrophil-specific myeloperoxidase promoter, which allows tracking of neutrophil movement to a tail cut injury. Compared to controls, 4 dpf BPA-exposed zebrafish show significantly reduced neutrophil recruitment to the site of injury (p=0.004). Control zebrafish receiving a tail cut injury at 4 dpf show a fourfold upregulation in CXCL8 expression one hour post-injury, whereas the BPA-exposed embryos display a blunted CXCL8 response. Because BPA-exposure at 0 dpf leads to a more than twofold increase in CXCR2 expression at 3 and 4 dpf compared to controls, neutrophil migration is likely reduced due to diminished CXCL8. Estradiol is known to reduce CXCL8 expression via the ER, and we found that BPA upregulates ERα and β gene expression by 3 dpf. During the first week of development, control zebrafish show an overall pattern of expression in all four tested genes that differs from that of BPA-exposed embryos. Our findings suggest that BPA leads to the overexpression of ERs, thus reducing CXCL8 expression and diminishing key protective innate immune responses.
Developmental and Environmental Regulation of Lung Macrophage Immune Response
Authors: Yamamura Asami , Lund Sean J , Lakhdari Omar , Sakai Mashito , Glass Christopher K , Prince Lawrence S 
Published: January 2023

Abstract
Neonates are uniquely susceptible to lung infection and injury. Alveolar (AM) and interstitial (IM) lung macrophages are the major cell populations protecting the lung against inhaled microbes and facilitating wound repair. AM and IM express unique cell surface markers and transcriptional profiles. We therefore hypothesized that neonatal and adult lung environments lead to differences in macrophage function. When lung macrophages were isolated by flow cytometry and stimulated with LPS in vitro for 2 h, neonatal AM expressed higher levels of Il1b and Il6 compared to adult AM. We next tested our hypothesis in vivo, challenging neonatal and adult mice with intranasal LPS. Contrary to our in vitro experiments, LPS induced higher cytokine expression in AM compared to IM. In addition, the adult AM response was higher than neonatal AM. To specifically test how the lung microenvironment regulated macrophage response, we stimulated neonatal and adult AM and IM with LPS either immediately after isolation or after 10–20 h of tissue culture. Consistent with our hypothesis, expression of key AM transcription factors fell quickly during cell culture. While neonatal AM and IM had similar LPS responses immediately after isolation, adult IM had higher cytokine expression compared to adult AM. Intriguingly, for both adults and neonates, longer culture time enhanced cytokine expression in activated AM while dampening the expression in IM. Our data suggest that the developmentally regulated lung microenvironment regulates the macrophage innate immune response.
Regulatory roles of suppressors of cytokine signaling 1 during type I vs type II macrophage polarization and phenotype determination
Authors: Lee Choong-eun , Jeong Hana , Kim Ga-young 
Published: January 2023

Abstract
During infection, activated macrophages display type 1 pro-inflammatory (M1) and type II anti-inflammatory (M2) features for pathogen clearence and the subsequent tissue repair, respectively. While suppressors of cytokine signaling (SOCS) play a role in the regulation of inflammation in diverse systems, the regulatory role of SOCS in M1 vs M2 activity has not been clearly established. We have studied SOCS action in macrophage phenotype determination in vitro employing SOCS gene transfer and knock-down using THP1 cells differentiated with PMA. For the LPS-induced M1 polarization, ROS signaling was crucial as the early ROS generation, MAPK activation, and the production of pro-inflammatory cytokines were all subject to inhibition by NAC, an anti-oxidant. Meanwhile, the dexamethasone (Dex)-induced M2 polarization involved ROS generation and MAPK/MPK1 activation in delayed kinetics, leading to the production of anti-inflammatory cytokines, which was also sensitive to NAC. Our previous studies demonstrated anti-oxidant functions of SOCS1 through the induction of thioredoxin and Nrf-2. In line with this, SOCS1 over-expression significantly inhibited M1 induction with ROS down-regulation, while SOCS1 ablation promoted ROS and pro-inflammatory/M1 cytokine production. In contrast, Dex-induced anti-inflammatory/M2 cytokines were severely reduced by SOCS1 knock-down with the impaired p38 and GILZ induction. The data support the anti-inflammatory action of SOCS1, while suggesting that SOCS1 exerts differential effects on M1 vs M2 polarization. The identification of distinct target molecules of SOCS1 action in M1vs M2 macrophage phenotype determination is under investigation.
Chronic Viral Infection Induces Metabolic Reprograming of Plasmacytoid Dendritic Cells
Authors: Greene Trever T , Jo Yeara , Zuniga Elina 
Published: January 2023

Abstract
Viruses that cause chronic infections, including Human Immunodeficiency Virus (HIV), Hepatitis B virus (HBV), and Hepatitis C virus (HCV), infect more than 500 million people worldwide and represent a major health burden. Chronic viral infections suppress immune responses to promote their own persistence. Plasmacytoid Dendritic Cells (pDCs) are an innate cell type which are specialized to produce exceptionally high levels of type I interferon (IFN-I) in response to viral infection, however in the context of chronic viral infection they lose the capacity to produce IFN-I. The metabolic program of pDCs has previously been implicated to be crucial for their function, but it is unclear how chronic infection modifies pDC metabolism. We demonstrate that chronic infection stimulates metabolic reprogramming in pDCs. Specifically, chronic infection is associated with sustained suppression of respiratory metabolism in pDCs. Furthermore, we demonstrate that respiratory metabolism in pDCs is highly correlated to their ability to produce cytokine, and that byproducts of respiratory metabolism (mitochondrial derived reactive oxygen species) can promote cytokine production in pDCs. These data suggest that chronic infection drives changes in pDC metabolism that could be associated with infection driven pDC exhaustion.
Clinical manifestations following intramuscular exposure of mice to a lethal dose of ricin
Authors: Sapoznikov Anita , Falach Reut , Gal Yoav , Evgy Yentl , Kronman Chanoch , Sabo Tamar 
Published: January 2023

Abstract
Ricin a plant toxin derived from seeds of Ricinus communis, irreversibly inactivates ribosomes by site-specific depurination, thereby arresting cell protein synthesis, while its clinical manifestations following intramuscular exposure have not yet been investigated in depth. Here we report an extensive pathological study of intramuscular intoxication with a lethal dose of ricin in a murine model.
Ricin injection caused neutrophilia, thrombocytopenia and coagulopathy. Although histopathological alternations including hemorrhages, vessel congestion and focal necrosis were observed in various organs, major insults were limited to the spleen, bone marrow (BM) and cardiovascular system. Intensive atrophy, evident in the BM and splenic white pulps, were accompanied be severe depletion of megakaryocytes in both organs. Moreover, splenic T and B lymphocytes, NK cells, macrophages and dendritic cells decreased significantly. The most striking damage was in the heart, where we observed diffuse hemorrhages, myocyte disconnection, loss of striation, interstitial edema, collagen deposition and elevated levels of serum cardiac troponin. Finally, echocardiography revealed marked thickening of the walls, along with sequential decrease in left ventricular volume. These were accompanied by a significant reduction in stroke volume and cardiac output, while the ejection fraction remained unaltered. Measurements of the toxin direct catalytic performance in the spleen, BM and heart of the ricin-intoxicated mice revealed no substantial 28S rRNA depurination. The lack of measurable ricin catalytic activity in conjunction with the striking clinical disorders observed, suggests that ricin affects these organs in an indirect manner.
NLRP12 regulates intestinal inflammation and tumorigenesis independent of gut microbiota
Authors: Khan Shahanshah , Zaki Hasan 
Published: January 2023

Abstract
Cytosolic NOD-like receptor NLRP12 plays a key role in suppressing intestinal inflammation and tumorigenesis via downregulation of NF-kB and MAPK pathways. Recent studies showed that altered gut microbiota in Nlrp12−/− mice is responsible for higher activation of NF-kB and MAPK pathways. Therefore, the precise function of NLRP12 in the intestinal remains poorly defined. Here we reinvestigated the contribution of gut microbiota in NLRP12-dependent regulation of colitis and colorectal cancer (CRC). We analyzed the microbiota composition of wild-type and Nlrp12−/− mice by 16S rRNA sequencing. Although there was a considerable difference in different microbial taxa between WT and Nlrp12−/− mice, no major difference was noted in Lachnospiraceae, which was previously been linked to hyperinflammatory responses in Nlrp12−/− mice. We next assessed the contribution of altered microbiota in colitis and CRC susceptibility of Nlrp12−/− mice. WT and Nlrp12−/− mice were co-housed before induction of experimental colitis and colorectal tumorigenesis. Separate or co-housed Nlrp12−/− mice developed significantly heightened colitis and increased burden of colorectal tumors. Higher activation of NF-kB, ERK, and STAT3 pathways was observed in both separate and co-housed Nlrp12−/− mouse colons during acute colitis. Further, macrophages from littermate or co-housed Nlrp12−/− mice exhibited hyperinflammatory responses along with higher activation of NF-kB and ERK pathways upon stimulation with LPS. Taken together, NLRP12 is an intrinsic regulator of NF-kB and MAPK pathways downstream of TLRs and suppresses inflammatory and tumorigenic responses in the intestine independent of gut microbiota.
MyD88 mediates the pathogenesis of dermatitis in SHARPIN-deficient mice by regulating TNF production on the skin
Authors: Sharma Bhesh Raj , Karki Rajendra , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
Mice deficient in SHANK-associated RH domain–interacting protein (SHARPIN), a component of the linear ubiquitin chain assembly complex (LUBAC), develop a spontaneous inflammatory disorder with pathologic hallmarks similar to atopic dermatitis and psoriasis in humans. Previous studies identified the crucial role of components of the TNF and IL-1 signaling pathways in the progression of disease in SHARPIN-deficient mice. However, an innate adaptor or sensor that relates to the disease progression has remained unknown. In this study, we found that the genetic ablation of myeloid differentiation primary response 88 (MyD88) completely rescued skin inflammation in SHARPIN-deficient (Sharpincpdm) mice. Systemic inflammation and immune cell dysregulation were also rescued. Fibroblasts derived from SharpincpdmMyd88−⁄− micefailed to provide protection against TNF-induced cell death. SharpincpdmMyd88−⁄− mice had reduced TNF production in their skin. Furthermore, depletion of the microbiota through the oral administration of antibiotics partially rescued both the skin inflammation and systemic inflammation, demonstrating a role for the gut microbiota in SHARPIN-deficient mice. Our findings suggest a detrimental role for the innate immune adaptor MyD88 in instigating skin inflammation in Sharpincpdm mice.