bluuebunny/crossref_metadata_2025_split huggingface dataset:
PD-1H facilitates phagocytic clearance of HIV infected T cells by monocyte derived macrophages and dendritic cells
Authors: Petersen Sean , Xu Xuequn , Camarillo Cynthia , Yi Guohua 
Published: January 2023

Abstract
Phagocytosis is a critical component of the immune response to viral infection, resulting in the clearance of infected cells, while minimizing exposure of uninfected cells. HIV is able to evade this response through suppression of key regulators of phagocytosis. Programmed death 1- homologue (PD-1H) has been identified as a co-inhibitory molecule that suppresses activation of T cells during infection and cancer immunogenesis. However, our evidence indicates that PD-1H may also play a significant role during immune response to HIV infection via upregulation and subsequent phagocytosis of infected T cells. HIV-induced apoptosis and monocytic cell engulfment was tested utilizing CEMSS T cells as target cells and monocyte derived macrophages (THP-1), as phagocytic cells. Cells were infected with a GFP-labeled HIV strain, NL4-3. HIV-infected T cells displayed greater apoptotic activity (approximately 9.0%) relative to mock-infected controls. Concurrently, phagocytosis of HIV-infected T cells increased approximately 4-fold. Expression of PD-1H on infected T-cells was detected on 16.7% of cells, which correlated with increased phagocytosis. When an antagonistic antibody against PD-1H was used, the number of phagocytic cells was reduced by a factor 2, which was replicated utilizing human stem cell derived dendritic cells. Phagocytosis was also confirmed by the upregulation of IL-1β, which was 5 fold higher relative to controls. Here we demonstrate that PD-1H facilitates monocytes-derived macrophages or DCs to phagocytize HIV-infected T cells in cell based assays. Based on these results we believe that further study in humanized mouse models of HIV infection is warranted.
Epithelial Cell Control of Adaptive Immune Plasticity in Periodontal Disease
Authors: Wallet Shannon , Delitto Andrea , Rocha Fernanda , Amador Byron , Sorenson Heather 
Published: January 2023

Abstract
In periodontal disease (PD), subgingival bacteria initiate and sustain a non-resolving inflammation that is ineffective at controlling the infection. T cells are critical for proper control of infection, but also homeostasis at mucosal surfaces including that of the periodontium. We have previously demonstrated that PD is associated with the local expression of cytokines associated with induction of Th(c)1/Th(c)17 plasticity, whereby loss of oral epithelial cell (OEC) MyD88-dependent-signaling results in exacerbation of this response. The objective of this study was to determine in which T-cell compartment plasticity occurs during PD and whether MyD88-dependent OEC signaling regulates this plasticity. Mice with MyD88-sufficient (B6) or MyD88-deficient (B6OEC.MyD88) OEC were subjected to oral polymicrobial infection for six weeks, after which the severity of disease as measured by alveolar bone loss and immunophenotype of the cellular infiltrate were evaluated. A temporal reduction in the frequency of Th(c)17 cells along with an increased frequency of Th(c)1 and Th(c)1/Th(c)7 cells was observed in B6 mice as PD progressed. This conversion was concomitant with a decrease in the frequency of regulatory T-cells and soft tissue IL10 and TGFβ expression. This immunophenotype was exacerbated in time of onset and magnitude in B6OEC.MyD88mice and was associated with an increase in alveolar bone loss. Together these data demonstrate that T-cell plasticity plays a role in PD whereby OEC-MyD88-signaling at least in part controls this plasticity. These data suggest that OECs can be a target for local modulation of T-cell plasticity as an intervention for the treatment of periodontal disease.
Characterizing the function of the CD300e immunoregulatory molecule.
Authors: Chen Kate , Silveira Pablo , Lo Tsun Ho , Hart Derek , Clark Georgina 
Published: January 2023

Abstract
The CD300 molecules are a family of immunoglobulin-like molecules that have immune regulatory functions. It includes both inhibitory and triggering members, which are highly conserved within the family as well as across species. CD300e, the most restricted member of the family, is expressed in low amounts in most of tissue, whereas its expression is upregulated in different cancers, such as, colorectal, stomach, liver, ovarian and pancreatic cancer. CD300e is highly expressed on CD14+ monocytes. Differentiation of monocytes into dendritic cells (DCs) and macrophages down regulated their CD300e expression. As no monoclonal antibodies are available yet to mouse CD300e, we generated a CD300e knock-out (KO) mouse as an alternative approach to study its biology. We have characterized the changes in immune cells populations resulting from CD300e deletion. The CD300eKO mice are in general fit and reproductively healthy. Mice with CD300e deficiency has a significantly reduction in their circulating leukocytes compared to wild type mice. Hematopoietic stem cells from CD300eKO mice were capable of differentiating in vitro into DC or macrophages with normal antigen uptake and T cell stimulatory capacity. We are using CD300eKO mice to elucidate CD300e function by further testing a variety of functions in monocytic cells.
Detection of chicken interleukin-23, a heterodimeric cytokine with monoclonal antibodies reacting p19 and p40 subunits
Authors: Kim Woohyun , Lim Yeaseul , Lillehoj Hyun S. 
Published: January 2023

Abstract
Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two covalently linked subunits, p19 (IL-23α) and p40 (IL-12/IL-23β) which is shared with IL-12. IL-23 plays an important bridging role between the innate and adaptive immune responses. Recently, chicken IL-23 (chIL-23) has characterized and shown functional similarity to mammalian IL-23 in production of IFN-γ from T cells and macrophages. In the present study, thus, we developed mouse monoclonal antibodies (mAbs) against two chIL-23 subunits, p19 and p40 and characterized their specificities in various immunoassays. To differentially detect IL-12 and IL-23 which share p40 subunit, both were expressed functionally by cotransfection with genes encoding IL-12α (also known as p35 subunit) and IL-12/IL-23β, or IL-23α and IL-12/IL23β into mammalian system. By several immunoassays including Western blot, and immunocytochemistry, several IL-23α mAbs react with IL-23 were found out. We also found some IL-12/23β mAbs react with IL-12 and IL-23 and some react with IL-23 only, but not with IL-12. Using these mAbs, we developed capture ELISA assay to quantify native IL-23 from various samples. One of the mAbs also exhibited its activity to neutralize the IL-23 in production of IFN-γ in chicken lymphocytes. We believe that these mAbs will be important immune reagents for basic and applied research of IL-23 in poultry.
Lipocalin2 mediates anti-inflammatory functions through the inhibition of STAT3 and activation of STAT5 signaling in bone marrow derived macrophages
Authors: samuels janaiya , Rayalam Srujana , Shashidharamurthy Rangaiah 
Published: January 2023

Abstract
Lipocalin2 (Lcn2), an innate immune protein, is known to be significantly up-regulated during inflammatory disorders including arthritis, however, its molecular mechanisms remain to be elucidated. We have shown that Lcn2-knockout (Lcn2KO) mice developed more severe serum-induced arthritis (STA) compared to wild-type (WT) mice with significantly reduced neutrophil infiltration but considerably more macrophage migration. Therefore, we have investigated the possible role of Lcn2 in regulating macrophages. Bone marrow cells were isolated from Lcn2KO and WT mice and polarized to classical or inflammatory (M1) and alternate or anti-inflammatory (M2) macrophages. The polarization of M1 (iNOS) and M2 (arginase-1) phenotypes was confirmed by Western blot analysis. We did not observe a difference in polarization of Lcn2KO and WT macrophages suggesting that Lcn2 may not alter the polarization of macrophages. However, anti-inflammatory cytokines such as TGF-β1 and IL-10 were significantly reduced in either LPS or IC-stimulated M2 phenotypes from Lcn2KO mice compared to WT mice. In agreement, we observed systemic elevated levels of pro-inflammatory cytokines in Lcn2KO compared to WT mice during STA. Further, we’ve demonstrated that Lcn2 deficient M1 macrophages displayed increased STAT3 activation compared to WT cells. In addition, WT M2 phenotype exhibited elevated STAT5 activation. Taken together, these results suggest that Lcn2 may promote the down-regulation of pro-inflammatory cytokines and up-regulate anti-inflammatory cytokines possibly through STAT3 and STAT5 signaling pathways as a potential negative feedback loop in order to limit the extent of inflammation during autoimmune arthritis conditions.
Intragenic transcriptional interference regulates the human immune ligand MICA.
Authors: O’Callaghan Chris , Lin Da , Hiron Thomas K. 
Published: January 2023

Abstract
Regulation of MICA expression is incompletely understood, but human MICA can be upregulated in cancer cells, virus-infected cells and rapidly proliferating cells. Binding of MICA to the activating NKG2D receptor on cytotoxic immune cells promotes elimination of the cell expressing MICA. We noted that MICA has tandem promoters that drive overlapping forward transcription. We show that the MICA gene contains a conserved upstream promoter that expresses a non coding transcript. Transcription from the upstream promoter represses transcription from the standard downstream MICA promoter in cis through transcriptional interference. The effect of transcriptional interference depends on the strength of transcription from the upstream promoter and quantitative studies show that it is described by a simple reciprocal repressor function. The time course of transcriptional interference coincides with recruitment at the standard downstream promoter of factors involved in nucleosomal remodeling during transcription. Transcriptional interference is demonstrated in the regulation of MICA expression by the physiological inputs interferon-γ and interleukin-4, that both act through regulatory DNA elements in the upstream promoter. These findings have significant implications for the understanding of MICA expression. Transcription factors activating the downstream promoter will upregulate MICA expression, whereas transcription factors activating the upstream promoter will downregulated MICA expression. A genome-wide analysis indicates that transcriptional interference between tandem intragenic promoters may be involved in regulating the expression of multiple other human genes.
Critical role of miR-23b in late sepsis-induced immunosuppression.
Authors: Yin Deling , Caudle Yi , Zhang Haiju 
Published: January 2023

Abstract
Sepsis is characterized with immunosuppression in late phase. microRNA-23b (miR-23b) is a multiple functional miRNA, which has been reported to prevent multiple autoimmune diseases through the regulation of inflammatory cytokine pathways. We hypothesize that miR-23b plays a role in the pathogenesis of sepsis. Our study verified the effect of miR-23b on sepsis induced immunosuppression. We found miR-23b expression was upregulated in early sepsis and sustained in late sepsis. The activation of TLR4/9/p38/STAT3 signal pathway contributes to the production of miR-23b in CLP-induced sepsis. Simultaneous in vivo blockade of miR-23b with inhibitor injection after sepsis initiation decreased TUNEL positive cells in spleen, improved survival via reducing the mortality by 42%. MiR-23b inhibitors restored the T cell immunoreactivity by alleviating the development of T cell exhaustion and producing smaller amounts of immunosuppressive interleukin 10 (IL-10) and IL-4 in late sepsis after LPS stimulation. We also demonstrated that miR-23b triggered apoptosis and T cell exhaustion to mediate immunosuppression in late sepsis by inhibiting the activation of non-canonical NF-κB signal and promoting pro-apoptotic signal pathway via targeting NIK, TRAF1 and XIAP. Circulating miR-23b serum levels might be a potential useful marker to predict survival in patients with critical sepsis. Blocking the expression of miR-23b may be an effective approach for prevention and treatment of sepsis-induced immunosuppression.
BCAP, a novel TLR-IL1R signaling adapter, regulates the differentiation of pathogenic Th17 cells
Authors: Deason Krystin L. , Troutman Ty Dale , Jain Aakanksha , Pasare Chandrashekhar 
Published: January 2023

Abstract
Toll-IL-1R homology (TIR) domains are found within adaptor proteins involved in the signaling of Toll like receptors (TLRs) and Interleukin 1 receptor (IL1R) families. Previous work by our lab identified a TIR domain in the protein B cell adaptor for phosphoinositide 3-kinase (BCAP) and determined a role for BCAP in TLR signaling in myeloid cells. Due to the shared use of TIR domains by TLR and IL1R signaling pathways, we hypothesized that BCAP would also be involved in signaling downstream of the IL1 family of receptors. The IL1 cytokine family has been shown to play a major role in T cell activation, survival, and differentiation; IL1b, in particular, plays a critical role in Th17 lineage cells. We have discovered that BCAP functions downstream of IL1R in CD4 T cells and regulates Th17 differentiation and function. IL1b-induced PI3K-Akt signaling was compromised in BCAP deficient T cells, which contributes to decreased mTOR activation, decreased glycolysis, and defective Th17 differentiation. Transcriptional analysis of BCAP deficient CD4 T cells led to the finding that BCAP is critical for the expression of genes associated with pathogenic Th17 lineage cells. Mice specifically lacking BCAP in T cells have normal development of steady state Th17 cells in vivo yet have decreased experimental autoimmune encephalomyelitis (EAE) disease. This study establishes BCAP as a critical link between IL1R and the metabolic status of activated Th17 cells and further demonstrates that BCAP is an important adapter molecule that functions downstream of IL1R and is critical for the generation of pathogenic Th17 cells in vivo.
Inhibition of Inositol-Requiring Enzyme 1 α in Established TH2 Cells Decreases the Secretion of Cytokines IL-5 and IL-13, but Not IL-4
Authors: Kemp Kyeorda L. , Poe Cody , Youngblood CheyAnne 
Published: January 2023

Abstract
TH2 cells are crucial in immune responses against extracellular pathogens; however, they can play a predominant role in a number of diseases due to inappropriate IL-4, IL-5, and IL-13 cytokine expression. Therefore, understanding how these cells develop and regulate their effector functions is of great importance. In response to an excess of unfolded proteins, serine-threonine kinase Inositol-Requiring Enzyme 1 α or IRE1α activates the unfolded protein response, UPR, which promotes increased protein folding capacity. T cell activation induces ER stress, and inhibition of IRE1α RNase activity in activated primary cells, via treatment of the cells with the commercially available drug 4μ8c, results in the reduction of IL-5, IL-4, and IL-13. Prior to this work, it was unknown if 4μ8c could inhibit TH2 cytokines in established TH2 cells. Treatment of a mouse TH2 cell line and differentiated human TH2 cells with 4μ8c results in inhibition of IL-5 and IL-13, but not IL-4 as measured by ELISA. The reduced cytokine expression is not due to differences in mRNA stability or mRNA levels; it appears to be due to a defect in secretion, as the cells appear to produce cytokines IL-5 and IL-13 as measured by flow cytometry. This suggests that the inhibition of these cytokines is due to post-translational processes. IL-5 and IL-13 produced by TH2 cells are critical for promoting chronic, inflammatory asthma, and 4μ8c blocks their expression in T cells in vitro. Future studies will determine if 4μ8c treatment can ameliorate the effects of these cytokines in a disease model. This work is supported by the NSU Faculty Research Council grant and OK-INBRE NIH P20GM103447.
Functional dissection of a novel IL-22 super-enhancer
Authors: Saini Ankita , Li Shasha , Bando Jennifer K. , Collins Patrick , Cella Marina , Colonna Marco , Oltz Eugene M. 
Published: January 2023

Abstract
Innate lymphoid cells (ILCs) are an emerging family of immune cells that serve essential roles in initiation, regulation and resolution of different immune responses. ILCs respond promptly to various signals present in their microenvironment by producing an array of cytokines and thereby directing the immune responses. These cells have been categorized as ILC1, ILC2 and ILC3 depending upon the effector cytokines secreted and transcription factors expressed by them. The signature effector cytokines produced by ILC3s are IL-22 and IL-17, providing protective immunity against extracellular pathogens. Another crucial facet of IL-22 function is its maintenance of mucosal homeostasis by modulating processes like tissue repair. However, there is a void of knowledge about genomic regions regulating IL-22 in different immune cells. In this regard, recent studies from our lab revealed a novel super-enhancer (SE), regulatory region composed of a large cluster of conventional enhancers, which likely regulates IL-22 and, perhaps, the neighboring IFNG gene. To dissect ILC3-specific functions of enhancers within the SE regulatory region, we are exploiting a murine cell line, MNK3, which mimics ILC3 functions in vitro and in vivo. We found that CRISPR-Cas9 mediated deletion of 5′ and 3′ end of this novel SE abrogated IL-22 expression in MNK3 cells. Furthermore, to identify cell and/or agonist specific elements of SE we set up a CRISPRi screen for which we generated a stable line of MNK3 expressing Dox inducible KRAB-dCas9. Together, our approach will define the cis-elements that regulate IL-22 in distinct immune cells and hence opening the field for novel therapeutic interventions for inflammatory diseases including autoimmune disorders.
Comparison of the open chromatin landscapes of effector and anergic T helper cells
Authors: Torres Annabel , Johnson Matthew , Wells Andrew D. 
Published: January 2023

Abstract
Costimulation through the TCR and CD28 drives T cell activation and differentiation. TCR engagement without CD28 results in anergy, a dominant state refractory to further stimulation. To understand how signals from CD28 and/or TCR imprint anergic and effector cell fates, we analyzed the genome-wide patterns of open chromatin in TCR or TCR/CD28-costimulated CD4+ T cells using ATAC-seq. Remarkably, although anergy represents a loss of T cell function, anergic cells exhibited 40% more open chromatin regions (OCR) than effector cells. Over 85% of the accessible effector genome was shared with anergic cells, and only 6,400 OCR were effector-specific. Conversely, only 60% of the accessible anergic genome overlapped with effectors, while 24,000 OCR were anergy-specific. Nearly one-fifth of effector-specific OCR were located in promoters, and were enriched for ETS, BCL6, and KLF consensus binding motifs. The remaining 5300 OCR, putatively representing the effector-specific enhancer landscape, were enriched for CTCF, ETS, BATF, IRF, and E2F motifs. Conversely, only 5% of anergy-specific OCR were in promoters, and these regions were enriched for motifs for FOS, ETS, IRF, SMAD3, and EGR1. The remaining 23,000 OCR that putatively constitute the anergy-specific enhancer landscape were specifically enriched for NFkB, TBET, NFAT, and Nur77 motifs. These results suggest that while CD28 cooperates with the TCR to activate a minor, effector-specific component of the T cell gene regulatory architecture, the major epigenomic role of costimulation is to actively oppose remodeling of the anergy-associated gene ‘regulome’ imprinted by default by the TCR.
T cell-independent mechanisms for protection against Mycobacterium tuberculosis
Authors: Izzo Angelo A. , Creissen Elizabeth , Izzo Linda , Troudt JoLynn , Bickett Thomas 
Published: January 2023

Abstract
Mycobacterium tuberculosis is responsible for millions of deaths each year and with the rise of drug resistance, the need for an effective vaccine is paramount. Currently, the Mycobacterium bovis strain, BCG is the only vaccine available, which provides inconsistent efficacy against pulmonary tuberculosis. In the mouse model, BCG induces adequate protection against M. tuberculosis infection, even in the absence of T cells. In our preliminary studies, subcutaneous BCG vaccination sufficiently protected mice when given 7 days prior to pulmonary infection with M. tuberculosis, reinforcing the T cell-independent nature of immunity and leading us to hypothesize that innate immunity was responsible. Various studies have defined a significant role for innate immunity during BCG vaccination in what has been termed “trained innate immunity”. However, our studies suggest that whereas “trained innate immunity” may play a role in BCG induced protection against non-mycobacterial diseases, this may not be the case for M. tuberculosis infection. We have found that the ability of BCG to replicate is key and that this may be due to its residence within phagocytic cells, as not all cells of the innate immune system are required for this outcome, based on selective cell depletion studies. As the effect of BCG on innate immunity is beneficial for other diseases, this study focuses on the early immune response shortly after BCG vaccination in the mouse model to help elucidate the mechanisms through which BCG stimulates protection and induces killing of M. tuberculosis. Our studies uncover novel mechanisms of BCG induced immunity that may be important to understanding new vaccination strategies.
CARDS toxin is critical for mediating the early inflammatory response during Mycoplasma pneumoniae infection
Authors: Segovia Jesus A. , Somorajan Sudha R. , Cagle Marianna P. , Baseman Joel B. , Kannan Thirumalai R. 
Published: January 2023

Abstract
Mycoplasma pneumoniae is a leading cause of bacterial community-acquired pneumonia and is implicated in initiation and exacerbation of asthma. One of the characteristic signs of M. pneumoniae infection in humans is an imbalanced proinflammatory immune response. A recently identified M. pneumoniae virulence factor is the Community-Acquired Respiratory Distress Syndrome (CARDS) toxin, an ADP-ribosylating and vacuolating protein produced abundantly during infection that is capable of displaying inflammation and pathology similar to infection with live M. pneumoniae. Our prior studies have focused heavily on the potent inflammatory effects of CARDS toxin in vitro, however its exact role during infection is unclear. We hypothesized that an M. pneumoniae strain lacking CARDS toxin expression would be unable to trigger the robust inflammatory response in vivo that is necessary for M. pneumoniae pathogenesis.
We genetically modified the clinical isolate M. pneumoniae S1 strain to generate a CARDS-deficient mutant we designated M7. M7 and S1 were used in vivo to identify changes in the proinflammatory response. Interestingly, we observed a drastic difference in the dynamics of M. pneumoniae infection during the early course of infection. Secretion of the cytokines TNF-α and CXCL1 were both significantly altered early during infection in M7-infected mice. The effects of this response led to a significant impact on Mycoplasma survival. By utilizing a novel CARDS toxin-deficient mutant of M. pneumoniae, our findings provide valuable insight into our understanding of the innate immune response against M. pneumoniae and implicate CARDS toxin as a potentially important target for therapeutic intervention during M. pneumoniae infection.