bluuebunny/crossref_metadata_2025_split huggingface dataset:
Altered colonic intraepithelial lymphocyte composition and function in Crohn’s Disease
Authors: Ohri Neha , Gerich Mark E , Fennimore Blair P , Kuhn Kristi 
Published: January 2023

Abstract
Inflammatory bowel disease (IBD), which is subdivided into Ulcerative Colitis (UC) and Crohn’s Disease (CD), is a chronic inflammatory disease of the intestine with an incompletely understood pathogenesis. Studies to date have indicated that gender, microbiome, epithelial biology, and host immune responses contribute to disease. Intraepithelial lymphocytes (IELs) are T cells within the intestinal epithelium that are in close contact with bacterial populations; they are prime candidates to study the interaction between intestinal microbiota and host immunity. Thus, we evaluated IELs in pinch biopsies of uninflamed colon tissue from subjects with biopsy-proven IBD and controls that were undergoing routine colonoscopies. Using flow cytometry, we observed a significantly increased percentage of TCRγδ+ CD8αα and CD44+ IELs in CD compared to controls, suggesting an increase in activated IELs in individuals with CD. Mitogen-stimulated IELs from subjects with CD showed significantly increased TNF-α secretion. Furthermore, while none of our controls secreted IL-17, a subset of CD patients, all premenopausal women, had IELs that produced large amounts of this cytokine. Thus, our data indicate differences in IEL populations between CD, UC, and controls, which support the pathophysiologic differences between CD and UC. The different phenotypes suggest a role for these cells in the diagnosis of IBD. Additionally, given the inflammatory cytokine profile of IELs in subjects with CD, especially women, we propose that IL-17+ IELs are an interesting population of effector T cells that deserve further study to increase our understanding of this complex disease.
DNA sensing via STING regulates immune responses in a lung cancer model via IDO
Authors: Mohamed Eslam , Lemos Henrique , Huang Lei , Ou Rong , Pacholczyk Gabriela , Arbab Ali , Munn David , Mellor Andrew 
Published: January 2023

Abstract
The Stimulator of Interferon Genes (STING) is a signaling adaptor associated with the endoplasmic reticulum. STING is activated by cytosolic DNA sensors to stimulate IFN type I expression (IFN-I). Recent reports revealed that DNA sensing to activate STING in dendritic cells (DCs) that engulfed dying tumor cells impeded growth of immunogenic tumors and potentiated responses to therapy. Consistent with these findings, synthetic STING agonists enhanced tumor regression. Paradoxically, Lewis Lung Carcinoma (LLC) grew slower in STING-deficient mice, revealing that STING is required for optimal LLC growth. Mechanistically, STING ablation abolished upregulation of the immunoregulatory enzyme, indoleamine 2,3 dioxygenase (IDO) in DCs located in tumor draining lymph nodes during LLC growth. Also, expression of the regulatory cytokine IL-10 and infiltration of myeloid derived suppressor cells (MDSCs) into the tumor microenvironment (TME) was diminished in mice lacking STING. In contrast, STING was not required for optimal growth of LLC tumors expressing neo-antigens, revealing a pivotal role for tumor antigenicity in influencing responses to DNA in the TME. These findings support the hypothesis that DNA from dying cells are sensed in the TME to activate STING, which induces dominant regulatory responses via IDO when tumor antigenicity is low and dominant immunogenic responses when tumor antigenicity is enhanced. Thus, cytosolic DNA sensing to activate STING has dichotomous roles in the TME, and tumor antigenicity is a pivotal factor influencing immune responses to DNA at sites of chronic inflammation associated with tumor growth.
Immune Enhancing Effects of Aqueous Extracted Black Raspberry in Diet-induced Hyperlipidemia Mice
Authors: Lee Youngjoo , Kim Jiyeon , An Jinho , Lee Sungwon , Lee Heetae , Kong Hyunseok , Song Youngcheon , Choi Hye Ran , Shin Daekeun , Kwon Ji-Wung , Kim Kyungjae 
Published: January 2023

Abstract
As obese population has increased today, metabolic diseases accompanied with obesity have been issued as well. Hyperlipidemia, one of those metabolic abnormalities, is commonly diagnosed with obesity. It is also associated with inflammation and immune dysfunctions, which can be led to global immune decline as the host ages. In this study, the potential effects of black raspberry on the diet-induced hyperlipidemia were examined in terms of immunology. We first induced hyperlipidemia in mice by feeding with 45kcal high fat diet (HFD) for 23 weeks and the progress of disease was traced by blood lipid profiling. Oral administration of black raspberry then proceeded for 16 weeks. As a result, the lymphocyte subset populations such as CD4+, CD8+ and CD11c+ were quantitatively restored by black raspberry treatment. The proliferation and cytokine production of lymphocytes were also normalized in black raspberry treated mice. Regarding specific immunity, oral administration of black raspberry significantly restored both cellular and humoral immune function as determined by in vivo cytotoxic T lymphocytes (CTLs) assay and specific immunoglobulin G (IgG) production level. Lastly the lipid indices including total cholesterol and HDL were considerably improved by black raspberry treatment, indicating the therapeutic effect of black raspberry on hyperlipidemia. That is, not only practical improvement in hyperlipidemia, but also the immune-enhancing effect of black raspberry was newly elucidated with quantitative and qualitative evidences.
Distribution of CD6 on B cells and its role in regulating B1a cell self-renewal
Authors: Enyindah-Asonye Gospel M , Li Yan , Singer Nora , Gupta Neetu , Fung John , Lin Feng 
Published: January 2023

Abstract
CD6 has been established as a marker and an important regulator for T cells. However, despite a previous report suggesting that CD6 is present on human B1a cells that are the primary source of natural antibodies, the precise distribution and potential role of CD6 on B cells remain unclear. In this report, by studying wild type and CD6 knockout (KO) mice, we found that murine CD6 is not detectable on any B2 cells, but is differentially expressed on B1a cells, and its expression is regulated by the activated state and anatomical location of the B1a cells. CD6 KO mice had significantly reduced numbers of B1a cells in different tissue compartments as a result of impaired ability to undergo self-renewal, and these mice had reduced levels of serum natural antibodies including levels of natural IgM. Consistent with previous reports showing that B1a cells produced natural IgM is critical in the pathogenesis of intestinal ischemia/reperfusion-induced injury, we also found that intestinal inflammation and tissue damage was significantly reduced in CD6 KO mice in this model. These findings clarified the distribution of CD6 on B cells and its expression regulation, and revealed a critical role of CD6 in regulating B1a function
Kinetic Measures of Macrophage Chemotaxis
Authors: Tikhonenko Maria , O’Clair Lindy , Roddy Meagan , Schroeder Kirk , Appledorn Daniel 
Published: January 2023

Abstract
Macrophages are crucial regulators of homeostasis, however, their functions can be compromised, leading to a wide variety of diseases such as diabetes, fibrosis, atherosclerosis and tumor metastasis. Macrophage recruitment by cytokines is an essential step of their function. The main obstacles for studying macrophage chemotaxis in vitro include low resident tissue macrophage cell number, lengthy isolation periods, and low percentage of migrating cells. In this report we illustrate a novel, fully automated, kinetic, label-free method for measuring macrophage chemotaxis. This method requires significantly fewer cells compared to traditional methods, thus allowing for increased throughput. Using the human monocyte cell line, THP-1, we first examined the directed migration of untreated vs. PMA-differentiated (macrophage-like) THP-1 cells. Using as few as 5000 cells per well, we observed a significantly different chemotactic response depending on treatment: pre-activated THP-1 had minimal to no response to C5a while differentiated THP-1 had a strong concentration-dependent response. Primary human PBMC-derived Macrophages were also evaluated. Using 2000 cells per well we found that both M1 (pro-inflammatory) and M2 (pro-regenerative) macrophages consistently responded to C5a with minimal, yet significant responses towards Rantes, SDF-1α, CXCL10, MDC and M-CSF. M1 cells exhibited slower response compared to M2 cells. Our data also revealed a significant alteration in kinetic responses depending on C5a concentration, with higher concentrations of C5a eliciting a slower response than lower concentrations, thus illustrating the requirement for kinetic measurements when investigating macrophage chemotaxis.
TSLP acts on neutrophils to drive complement-mediated killing of methicillin-resistant Staphylococcus aureus
Authors: West Erin E , Spolski Rosanne , Kazemian Majid , Kemper Claudia , Leonard Warren J 
Published: January 2023

Abstract
Community-acquired methicillin-resistant Staphylococcus aureus (MRSA) infections, often presenting as serious skin infections in otherwise healthy individuals, have become a world-wide epidemic problem and warrant attention for therapeutic intervention. Accordingly, defining the mechanisms that govern the activation and regulation of the immune response to MRSA is clinically important and could lead to the discovery of much needed rational targets for therapeutic manipulation. The cytokine thymic stromal lymphopoeitin (TSLP) is most well characterized as a major contributor to Th2-type responses in which it plays a deleterious role in the promotion of allergic responses. However, while TSLP is highly expressed at barrier surfaces, including the skin, its role in host-defense against bacterial infections has not been well elucidated. Unexpectedly, we found that TSLP can play a protective role in the skin by acting directly on mouse neutrophils to increase their killing of MRSA in both an in vitro killing assay and an in vivo skin infection model. Additionally, TSLP also directly acts on purified human blood neutrophils to reduce MRSA burden. Therefore, neutrophils represent previously unrecognized potent responders to TSLP. Surprisingly, we found that TSLP mediates these effects by directly engaging the complement C5 system to modulate reactive oxygen species production by neutrophils. Thus, TSLP increases MRSA killing in a neutrophil- and complement-dependent manner, revealing an unanticipated key connection between TSLP and the innate complement system. These data demonstrate an unrecognized beneficial role for TSLP in the skin, with potentially important clinical implications.
Human neutrophil extracellular trap formation is modulated upon stimulation with Toll-like-receptor ligands
Authors: Singh Ram Raj , Randhawa Darshan , Valera Isela 
Published: January 2023

Abstract
Polymorphonuclear neutrophils are considered to be the first line of defense against infections. One of the many strategies that neutrophils use for removal of foreign material is via the deployment of neutrophilic extracellular traps (NETs). NETs are chromatin fibers decorated with antimicrobial proteins. It is presumed that all NETs produced by neutrophils (mainly through stimulation with PMA) are similar in function and structure. Here, we demonstrate that when neutrophils are stimulated with different Toll-like-receptor (TLR) ligands, they produce NETs which are unique in structure and function with respect to its TLR stimulation. Not only does this demonstrate at the physiological level that NET formation is selective to its TLR Ligand, but it also represents a new layer of neutrophil NETosis modulation. Further elucidation of this layer of modulation of NET formation by endogenous and exogenous TLR ligands in disease states may have important implications for the pathogenesis and intervention in human diseases.
Innate IFNs and pro-inflammatory cytokines in local control of Respiratory Syncytial Virus infection of respiratory epithelial cells.
Authors: Uehling Megan , Hillyer Philippa , Shepard Rachel , Sheik Faruk , Luongo Cindy , Buchholz Ursula , Collins Peter L , Donnelly Raymond P , Rabin Ronald L 
Published: January 2023

Abstract
The host innate response contributes to the severity of disease caused by Respiratory Syncytial Virus (RSV). The primary target of RSV is the airway epithelium. The host becomes infected by inhaling droplets containing viral particles, which then infect small foci of cells. Infected cells then express types I and III interferons (IFN) and pro-inflammatory cytokines to alert neighboring epithelial and myeloid cells of danger. Despite their inherent differences, A549 (carcinoma, type II alveolar epithelium) and BEAS-2B (transformed bronchial epithelium) cells are often used interchangeably to study RSV-epithelial interactions. We compared these two cell lines by infecting them with low MOIs of RSV expressing GFP (rgRSV). We measured expression of types I and III IFNs, interferon stimulated genes (ISGs), pro-inflammatory cytokines and signaling intermediaries. We found that BEAS-2B cells contained rgRSV within foci of ~10–15 cells, but all A549 cells were infected by 48h. Both cell lines highly expressed IFN-β, IFN-λ1 and -λ2, but expression was greater in the A549s. Despite lower levels of IFN expression, BEAS-2Bs expressed higher levels of most classic antiviral ISGs (ISG15, MX1, PKR) and critical TLRs, RLRs and IFN receptors. In contrast, A549 cells expressed higher levels of NF-kB associated genes (CCL2, CCL5, CXCL8). Our data suggests that a balance between expression of NF-kB genes and ISGs may determine local control of RSV infection by respiratory epithelial cells.
A surprising link between the inflammasome and the centrosome
Authors: Davis Michael Alan , Fairgrieve Marian , Gale Michael 
Published: January 2023

Abstract
The NLRP3 inflammasome is a multisubunit protein complex of the innate immune system that senses and responds to a broad range of cell-intrinsic, pro-inflammatory stimuli. Upon activation in non-mitotic cells, NLRP3 aggregates with the inflammasome subunits ASC and Caspase-1 to assemble a single perinuclear superstructure. However, the processes and mechanisms that orchestrate inflammasome assembly and drive pyroptosis are largely unknown. Here, we show that the inflammasome recruits and colocalizes with known centrosome-associated proteins, γ-Tubulin and Centrin, in a Caspase-1 dependent manner. While the NLRP3 inflammasome does not colocalize with the centrosome itself, we show that the inflammasome assembles in close proximity to the centrosome early after NLRP3 activation. Furthermore, inflammasome formation results in a breakdown of the microtubule network as well as decreased γ-Tubulin and Pericentrin staining at the centrosome. These observations are strengthened by our proteomic studies identifying &gt;30 centrosomal interacting with NLRP3 and/or Caspase-1. One protein in particular is Nek7, a serine/threonine kinase required for the recruitment of γ-Tubulin and Pericentrin to the centrosome, suggesting that inflammasome activation phenocopies Nek7 loss. Finally, depletion of Nek7 inhibits pyroptotic cell death. Therefore, these data reveal a novel link between the inflammasome and centrosome, possibly bridged by NEK7, and provide mechanistic insight into inflammasome formation and pyroptosis.
Nek7 is an essential mediator of NLRP3 activation downstream of potassium efflux
Authors: He Yuan , Zeng Melody Yue , Yang Dahai , Motro Benny , Nunez Gabriel 
Published: January 2023

Abstract
Inflammasomes are intracellular protein complexes that drive the activation of inflammatory caspases. The NLRP3 inflammasome has been implicated in the pathogenesis of several acquired inflammatory diseases as well as Cryopyrin-associated periodic fever syndromes (CAPS) caused by inherited NLRP3 mutations. Despite extensive investigation, the molecular mechanism leading to NLRP3 activation remains unknown. We have identified Nek7 as an NLRP3-binding protein that acts downstream of potassium efflux to regulate NLRP3 oligomerization and activation. In the absence of Nek7, caspase-1 activation and IL-1β release were abrogated in response to signals that activate NLRP3, but not NLRC4 or AIM2 inflammasome. NLRP3 associated with the catalytic domain of Nek7, but the catalytic activity of Nek7 was dispensable for activation of the NLRP3 inflammasome. Activated macrophages formed a high-molecular-mass NLRP3-Nek7 complex, which along with ASC oligomerization and ASC speck formation were abrogated in the absence of Nek7. Nek7 was required for macrophages harboring the CAPS-associated NLRP3R258W activating mutation to activate caspase-1. Mouse chimeras reconstituted with wild-type, Nek7−/− or Nlrp3−/− hematopoietic cells revealed that Nek7 was required for NLRP3 inflammasome activation in vivo. Our studies demonstrate that Nek7 is an essential protein that acts downstream of potassium efflux to mediate NLRP3 inflammasome assembly and activation.
Dysregulated type I interferon and inflammatory monocyte-macrophage responses cause lethal pneumonia in SARS-CoV-infected mice
Authors: Channappanavar Rudragouda , Fehr Anthony , Vijay Rahul , Jincun Zhao , Meyerholz David K , Perlman Stanley 
Published: January 2023

Abstract
Highly pathogenic human respiratory coronaviruses cause acute lethal disease characterized by an exuberant inflammatory response and severe lung damage. Although fatal outcomes due to immunopathological events following SARS-CoV infection have been well established, factors initiating detrimental inflammatory responses are not well understood. Since persistent elevation of IFN-I suggested a pathogenic role in SARS patients, we explored the possibility that IFN-I was critical in the initiation of events that led to lethal lung immunopathology. Using mice infected with SARS (Severe Acute Respiratory Syndrome)-CoV, we show that robust virus replication accompanied by dysregulated type I interferon (IFN-I) signaling orchestrated exuberant inflammatory responses and lung immunopathology with diminished survival. Delayed IFN-I signaling promoted the accumulation of pathogenic inflammatory monocyte-macrophages (IMMs) resulting in elevated lung inflammatory cytokine/chemokine levels, extensive vascular leakage and impaired virus-specific T cell responses. Genetic ablation of the IFN-αβ receptor (IFNAR) or depletion of IMMs completely protected mice from lethal infection, without affecting viral load. Together, these results demonstrate that IFN-I signaling is detrimental in highly susceptible SARS-CoV-infected BALB/c mice, in large part by promoting the influx of highly pathogenic inflammatory monocytes-macrophages (IMMs) and identify IFN-I and IMMs as potential therapeutic targets in patients infected with severe CoV and perhaps other highly pathogenic respiratory viruses.
Cytokine production in the skin is regulated by metabolites from the microbiome that influence histone acetylation.
Authors: Sanford James Asbury , Huang Chun-Ming , Gallo Richard L 
Published: January 2023

Abstract
Immune functions of the skin, including cytokine production from epidermal keratinocytes, are influenced by the resident microbiota. We hypothesized that a prominent member of the normal skin microbiome, Propionibacterium acnes (P. acnes), can affect cutaneous inflammation through the epigenetic mechanism of inhibiting histone deacetylase (HDAC) activity. GC-MS analysis revealed that P. acnes can generate high levels of the short-chain fatty acids (SCFAs) propionic and valeric acid. These molecules increased levels of histone acetylation as seen by Western blot analysis of H3K9ac and H3K27ac and ChIP-PCR of H3K9ac in the IL-6 promoter. Using qPCR, RNA-sequencing, and multiplex ELISA we found that HDAC inhibition alone had no effect on cytokine production from primary cultures of normal human keratinocytes (NHEK), but greatly increased the production of many cytokines and chemokines (i.e. IL-1β, IL-6, IL-8, TNFα, and CCL5) elicited by ligands of TLR2 or TLR3. In direct contrast, HDAC inhibition suppressed TLR-mediated inductions of these cytokines in monocytes and dendritic cells. The mechanism of inhibition in PBMCs and DCs was consistent with previous findings demonstrating increased expression of Mi-2β and activation of the NuRD co-repressor complex, events that were absent in NHEK. Inhibition of HDAC activity in mouse skin confirmed these responses in vivo. siRNA screening identified a novel mechanism involving HDAC8 and HDAC9 as responsible for these effects in keratinocytes. Together, these results show that the skin local environment and microbe-derived SCFAs influence inflammatory gene expression in a cell type-specific manner, thus explaining selective inflammation in diseases such as acne.
Engineered C-reactive protein with improved protective action against pneumococcal infection
Authors: Ngwa Donald Neba , Gang Toh B , Singh Sanjay K , Agrawal Alok 
Published: January 2023

Abstract
The anti-pneumococcal function of native C-reactive protein (CRP) involves the binding of CRP to phosphocholine molecules present on Streptococcus pneumoniae and subsequent activation of the complement system. However, when pneumococci recruit complement inhibitory protein factor H on their surface, they escape the attack of complement. We have reported previously that CRP, in a non-native conformation, expresses the ability to bind immobilized factor H. Accordingly, we hypothesized that such non-native CRP would bind to factor H on pneumococci in vivo, mask its complement inhibitory activity, and therefore should be able to protect against complement-resistant pneumococci. We performed site-directed mutagenesis of CRP and identified a mutant which bound to factor H-coated pneumococci but not to factor H in fluid phase. Mutant CRP was found to be free in the mouse serum and blood and its rate of clearance was similar to that of native CRP. As reported previously, native CRP protected mice from lethal pneumococcal infection when injected into mice 30 minutes prior to the administration of pneumococci, but did not offer any protection when injected at 24 hours after the administration of pneumococci. In contrast, that was not the case with mutant CRP; mutant CRP protected mice from infection more effectively than native CRP, by increasing the survival and decreasing bacteremia. We conclude that, while native CRP is protective only against early-stage infection, mutant CRP capable of binding to factor H may protect against both early-stage and late-stage infections and thus is superior to native CRP in fighting against the disease.