bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of PI3-Kinase signalling in immunomodulation by a protective innate defence regulator peptide (135.6)
Authors: Madera Laurence , Nijnik Anastasia , Waldbrook Matt , Mookherjee Neeloffer , Gardy Jennifer , Falsafi Reza , Elliott Melissa , Hancock Robert EW 
Published: January 2023

Abstract
The rise of antibiotic resistance has lead to the search for new therapeutics against bacterial infections. Innate defence regulators (IDRs), synthetic versions of natural host defence peptides, are being developed as anti-infective agents due to their immunomodulatory properties. HDPs have been shown to suppress infections by boosting leukocyte recruitment, while limiting harmful inflammation. To optimize the IDR regulatory function, we developed peptide 1002, a bactenecin derivative, by screening a peptide library for enhanced leukocyte chemoattractants induction in human blood mononuclear cells (hBMC). 1002 enhanced protection in a Staphylococcus aureus murine infection model, correlating with increased chemokine production and leukocyte recruitment. 1002 also suppressed inflammatory cytokine production by lipopolysaccharide-stimulated hBMC. IDRs are being deciphered to assist in their development as therapeutics. We found that 1002 regulation is dependent on the PI3-Kinase pathway, a signal cascade that regulates immune and inflammatory responses. PI3-K inhibition abrogated 1002-mediated chemokine induction and reduced its anti-inflammatory properties. 1002 influence on PI3-K signalling was confirmed by examining responses of pathway elements and by systems biology microarray analysis of the immune network by InnateDB. Understanding the mechanisms of IDR immune regulation via PI3-K modulation will aid in their development as novel anti-infective agents. This work was supported by Genome BC and Genome Prairie for the Pathogenomics of Innate Immunity Research Program, and by FNIH and CIHR through the Grand Challenges in Global Health Initiative.
The inflammasome as a therapeutic target to improve outcomes after injury to the central nervous system (135.73)
Authors: de Rivero Vaccari Juan Pablo , Lotocki George , Dietrich W. Dalton , Keane Robert W. 
Published: January 2023

Abstract
The inflammasome constitutes one of the host's defenses against pathogens and deleterious stimuli by activating caspase-1. Inflammation with production of cytokines processed upon inflammasome activation is a crucial part of the secondary cascade of cell death induced after central nervous system (CNS) injury. The molecular mechanisms underlying trauma-induced inflammation in the CNS are poorly defined. In this study, the cellular distribution and protein association of inflammasome components were investigated in spinal cords and brains of rats subjected to moderate cervical injury and fluid-percussion brain injury, respectively. Coimmunoprecipitation experiments revealed that in the CNS, Apoptosis-Associated Speck-like protein containing a CARD (ASC) associates with caspase-1, caspase-11, NLR family, Pyrin domain containing 1 (NLRP1) and the inhibitor of apoptosis XIAP, forming the NLRP1 inflammasome. Injury to the CNS affects the composition of the inflammasome resulting in increased association of caspases-1 and -11 and NLRP1 with ASC and cleavage of XIAP. Immunohistochemistry revealed that motor neurons and cortical neurons express the NLRP1 inflammasome proteins. Therapeutic neutralization of ASC after injury inhibited inflammasome activation as shown by decreased caspase-1 activation and XIAP cleavage, resulting in reduced lesion volume and improved functional outcomes after spinal cord injury. Thus the inflammasome forms part of the innate immune response in the
CNS and its inhibition may offer a novel therapeutic strategy to improve outcomes after injury and CNS inflammatory diseases.
This research was supported by the NIH/NINDS and the US Army.
Catecholamine and PKA-dependent regulation of Foxo1 mRNA and protein in S49 lymphosarcoma cells (136.12)
Authors: Davis Daniel H. , Cohen Rhonna , Chambers Donald A. 
Published: January 2023

Abstract
We previously linked psychological stress to lymphocyte physiology by demonstrating the presence of a norepinephrine (NE)/beta-2-adrenergic receptor/cAMP/ PKA-dependent mRNA decay system in T lymphocytes. Foxo1 and the PKA pathway can participate in the promotion of cell cycle arrest and apoptosis of T lymphocytes, so we explored whether a PKA-selective cAMP analog (8CPT-cAMP) or NE could regulate BALB/c murine thymomas, S49wt and S49kin- (PKA-deficient), by affecting the genetic expression of Foxo1. Real-time PCR studies showed a 2-3 fold upregulation of Foxo1 mRNA in S49wt, but not S49kin- cells with NE or 8CPT-cAMP 2hr after treatment. Foxo1 total protein, determined by immunoblotting, was increased 6hr after treatments. Actinomycin D treatment of S49wt cells incubated with NE or 8CPT-cAMP revealed that the increase in Foxo1 was not associated with altered mRNA stability. Transfection of S49wt and S49kin- cells with a Foxo1 promoter luciferase reporter suggested Foxo1 mRNA increases due to enhanced Foxo1 message transcription. These results associate NE/cAMP/PKA in modulating increased Foxo1 mRNA and protein expression, and links Foxo1 to PKA-mediated gene regulation of lymphocyte physiology.
Catecholamine/cAMP regulation of Thy1 mRNA stability in S49 cells involves a 3’UTR cis-acting element (136.13)
Authors: Koduvayur Sujatha P , LaJevic Melissa D , Cohen Rhonna L , Chambers Donald A. 
Published: January 2023

Abstract
Under conditions of psychological stress, release of the catecholamine, norepinephrine (NE), by sympathetic nerves has been reported. We described Thy1 mRNA regulation via a NE-ß2 adrenergic receptor-cAMP-PKA dependent pathway (NE pathway) in murine thymoma (S49) cells. To understand the molecular mechanism(s) underlying this regulation we studied the involvement of a 116nt region containing a putative ARE in the 3'UTR of Thy1 mRNA. The Thy1 ARE region was inserted downstream of the GFP coding region and the resultant plasmid was transfected into S49 cells. The presence of this region reduced the expression of exogenous GFP mRNA upon stimulation of the NE pathway. This ARE region also binds to known ARE binding proteins (bps) including AUF1, HUR, TIAR and Hsp70 and binding of ARE bps to known AREs like TNFα mRNA is competed away by the Thy1 ARE region. Additionally, silencing the ARE bp HUR, a known enhancer of mRNA stability, increased Thy-1 mRNA decay upon stimulation of the NE pathway in S49 cells. These results suggest that Thy1 mRNA UTR can bind RNA bps and contains a novel cis-acting, 116 nt. ARE region that participates in the mechanism of NE pathway regulated mRNA decay. Supported by NIH.
PRDI-BF1/Blimp-1 Associates with Cytokine-Mediated Activation of Human Natural Killer Cells (136.25)
Authors: Smith Matthew A , Wei Sheng , Djeu Julie , Caligiuri Michael , Wright Kenneth L 
Published: January 2023

Abstract
PRDI-BF1 (Positive Regulatory Domain I - Binding Factor 1,) is a zinc finger transcription factor, originally identified as a suppressor of interferon-beta in virally-infected human fibroblasts. It is most widely recognized as a key regulator of B-cell terminal differentiation into antibody-producing plasma cells. More recently, several reports have described functions in T-cell homeostasis. Here, we demonstrate that PRDI-BF1 is expressed in human natural killer (NK) cells and is involved in the attenuation of activation-induced cytokine gene transcription. Ex vivo stimulation of negatively-selected human CD56+ lymphocytes with activating cytokines increases levels of PRDI-BF1 mRNA and protein. This induction occurs primarily in the CD56dim population. The combination of IL-2, IL-12 and IL-18 leads to synergistic increases in PRDI-BF1 and IFN? as part of a global transcriptional profile that we have characterized via microarray analysis. Blockade of PRDI-BF1 induction via siRNA leads to enhanced TNFα and IFNγ transcription. Chromatin immunoprecipitation analysis of both NK cell lines and primary CD56+ lymphocytes reveals occupancy of several promoters, including IFNγ. Collectively, these data provide evidence for novel contributions of PRDI-BF1 to natural killer cell function and innate immunity.
IL-10 produced by host animals, but not by transduced donor B cells, is required for tolerance delivered by B-cell gene therapy (136.34)
Authors: Su Yan , Noben-Trauth Nancy , Scott David W. 
Published: January 2023

Abstract
Our previous studies indicated that activated B cells, which were retrovirally transduced with an IgG-antigen fusion construct, could effectively induce antigen-specific tolerance in vivo. Since IL-10 is an important cytokine for maintaining immune tolerance and suppression, in this study we investigated the role of IL-10 in the B-cell delivered gene therapy for tolerance induction. First we transduced either wild type (wt) or IL-10 knockout (KO) B cells with ovalbumin (OVA) peptide-IgG fusion protein via a retroviral vector and found that both wt and IL-10 KO B cells are tolerogenic antigen-presenting cells (APC) capable of inducing antigen-specific tolerance in host animals.. Next we tested whether IL-10 produced by host animals is required for tolerance induction. Animals that received OVA peptide-IgG B cells were injected weekly with anti-IL-10 receptor antibody (1B1.3a) or a control antibody (rat IgG1 GL113). This in vivo anti-IL-10 receptor blockade completely abrogated the tolerogenic effects of OVA peptide-IgG B cells. Together, our results suggest that tolerance induced by B cells requires the IL-10 produced by host, but not by donor B cells. These data suggest that regulatory T cells which produce IL-10 are involved in the tolerance induction by this B-cell gene therapy protocol. (Supported by NIH grants RO1 AI035622 and DK068343)
Th17 cells mediate sustained autoimmune inflammation and are highly resistant to restimulation-induced cell death (137.17)
Authors: Shi Guangpu , Ramaswamy Madhu , Vistica Barbara P. , Tan Cuiyan , Wawrousek Eric F. , Siegel Richard M. , Gery Igal 
Published: January 2023

Abstract
Both Th1 and Th17 T-cell subsets can mediate inflammation, but the kinetics of the pathogenic process mediated by these two subsets has not been investigated. Using an experimental system in which TCR-transgenic Th1 or Th17 cells specific for hen egg lysozyme (HEL) induce ocular inflammation in recipient mice expressing eye-restricted HEL, we found important differences in the in vivo behavior of these two subsets. Th1 cells initially proliferated considerably faster and invaded the eye more quickly than their Th17 counterparts, but then disappeared rapidly. By contrast, Th17 cells accumulated and remained the majority of the infiltrating CD4 cells in the eye for at least 15 days after transfer, mediating more long-lasting pathological changes. Th17 cells were highly resistant to restimulation-induced apoptosis, a major pathway by which autoimmune and chronically restimulated Th1 cells are eliminated. Th17 cells had reduced FasL production and resistance to Fas-induced apoptosis relative to Th1 cells despite normal surface expression of Fas. Moreover, Th1 recipient eyes had a higher expression level of FasL than Th17 recipient eyes. Resistance to programmed cell death in Th17 cells may enable more long-lasting responses against pathogens, but may also enable the chronic autoimmune tissue destruction that has been ascribed to Th17 cells.
Gender-dependent differential antigen uptake by CD11bloF4/80lo macrophages from experimental autoimmune encephalomyelitis resistant male and susceptible female SJL mice (137.2)
Authors: Hussain Shabbir , Stohlman Stephen A 
Published: January 2023

Abstract
Macrophages (MΦ) preferentially regulate induction of antigen-specific Th1 and Th2 immune responses to protein antigens in female and male SJL mice, respectively. Alexa Flour 488 conjugated ovalbumin (OVA) and self-quenched DQ-OVA that fluoresces following lysosomal degradation, were used to determine gender-dependent differences in antigen uptake and processing in peritoneal MΦ. Antigen uptake and processing are higher in CD11bhiF4/80hi compared to the CD11bloF4/80lo MΦ of both genders. CD11bhiF4/80hi MΦ in both genders exhibit similar levels of antigen uptake, processing and MHC class II expression. By contrast, CD11bloF4/80lo MΦ from males show increased antigen uptake and decreased MHC Class II expression compared to female CD11bloF4/80lo MΦ. Despite differences in antigen uptake and MHC class II expression, CD11bloF4/80lo MΦ from both genders show similar kinetics of antigen processing. CD11bloF4/80lo MΦ from both genders presented antigen to CD4+ T cells; however, increased IL-10 was induced during T cell activation by male CD11bloF4/80lo MΦ compared to female CD11bloF4/80lo MΦ. These data demonstrate increased antigen uptake, diminished Class II expression and preferential IL-10 secretion by CD11bloF4/80lo MΦ derived from Th2 responsive EAE resistant male SJL mice. This work was supported by National Institute of Health grant NS53824.
Human keratinocytes release and respond to danger signals in cutaneous lupus erythematosus (137.33)
Authors: Wittmann Miriam , Wang Dong , Eiz-Vesper Britta , Werfel Thomas 
Published: January 2023

Abstract
Lupus erythematosus (LE) is a chronic multisystem disease and cutaneous manifestations belong to the most common clinical features. It was the aim of this study to further decipher the role of keratinocytes in autoinflammation. In immunohistochemistry we found a high expression of inducible HSP70, IL-18R and IL-18 in lesional LE skin. Increased expression of TNFalpha and high mobility group box-1 (HMGB-1) has been described previously. Studies with cultured, patient derived keratinocytes showed that IL-18 stimulation results in significant TNFalpha production in LE derived but not in healthy keratinocytes. Furthermore, we could show that keratinocytes release high levels of HSP70 into the supernatant which was not due to apoptosis. Here we provide evidence, that HSP70-peptide complexes are internalised by keratinocytes and that antigenic peptides induce the production of IFNgamma in T cells via this route. Of note, HMGB-1 as well as TNFalpha significantly enhanced the uptake of HSP70 into human primary keratinocytes. In conclusion, we provide evidence that skin resident cells play an active part in maintaining LE inflammation also by means of secretion of molecules which belong to the danger-associated molecular pattern (DAMP) family. This study was supported by DFG grant Wi1822/5-1 and SFB566/A6.
Residual T cell potential in common myeloid progenitors (138.14)
Authors: Chi Anthony WS , Bhandoola Avinash 
Published: January 2023

Abstract
Downstream of hematopoietic stem cells (HSCs) and multipotent progenitors (MPPs), common myeloid progenitors (CMPs) are thought to represent a progenitor population that contributes to the development of myeloid and erythroid cells. CMPs do not contribute to T cell development in mice, as upon intrathymic transfer they fail to generate T cells in the thymus. However, when co-cultured on OP9 stromal cells expressing Delta-like Notch ligands, CMPs generated T cells, though less efficiently than HSCs or MPPs. Such inefficiency was demonstrated by the increased sensitivity of CMPs to GSI blockade. This defect correlated with reduced expression of Notch1 and also Lunatic fringe. Consistent with this finding, overexpression of ICN1 to mimic constitutively active Notch1 signaling was sufficient to drive CMPs into the T lineage developmental pathway in vivo, and resulted in development of T cell leukemia. This study shows CMPs are still receptive to Notch1 signaling and Notch-mediated T cell leukemogenesis. We are currently exploring mechanisms that might signal downregulation of Notch1 and Lunatic fringe in CMPs, and so initially constrain T potential. We are also reconstituting expression of Notch1 and Lunatic fringe in CMPs using retroviruses, and will determine whether such manipulations are sufficient to confer T potential to CMPs in vivo, or whether additional mechanisms that constrain T potential exist.
Characterizing Thymocyte Migration in the Medulla Using Two-Photon Microscopy (138.20)
Authors: Ladi Ena , Le Borgne Marie , Liao Ying Fang , Robey Ellen 
Published: January 2023

Abstract
To establish tolerance, a developing thymocyte must be screened for self-reactivity before exiting the thymus. However, the mechanism by which a thymocyte searches the medulla and antigen-presenting cells for self-antigens is not clear. We have developed a cut thymic lobe preparation that allows us to document thymocyte migration within the medulla by two-photon laser scanning microscopy. Medullary thymocytes migrate rapidly through confined random walk making short and dynamic contacts with dendritic cells. In response to negative selection signals, thymocyte migration is significantly slower and constrained to smaller areas. In addition, self-reactive thymocytes engage in long lasting contacts with dendritic cells. Polyclonal medullary thymocytes consist of a small but significant population of relatively slow thymocytes with more constrained migration that may correspond to auto-reactive thymocytes undergoing negative selection. Taken together, our data defines scanning strategies used by medullary thymocytes during negative selection and suggests that a significant proportion of medullary thymocytes are actively receiving negative selecting signals.
Egr1-Rcan1 axis serves as a molecular switch from activation to inhibition in IgE-mediated signalling (139.3)
Authors: Lin Tong-Jun , Yang Yong Jun , Molkentin Jeffery D , Berman Jason N 
Published: January 2023

Abstract
Mast cells play a central role in allergy. Aggregation of FcεRI activates a cascade of signaling events leading to mast cell activation. Subsequently, inhibitory signals are switched on for turning off activated signals. Here we identified that Rcan1 serves as a negative regulator for turning off FcεRI-mediated mast cell activation. FcεRI-induced Rcan1 expression was identified by suppression subtractive hybridization and verified by real-time quantitative PCR and Western blotting. Deficiency of Rcan1 led to increased calcineurin activity, increased NFAT and NF-κB activation, increased cytokine production and enhanced IgE-mediated late phase cutaneous reactions. Forced expression of Rcan1 in wild type or Rcan1 deficient mast cells reduced FcεRI-mediated cytokine production in vitro. Rcan1 deficiency also led to increased FcεRI-mediated mast cell degranulation and enhanced passive cutaneous anaphylaxis. Analysis of the Rcan1 promoter identified a functional Egr1 binding site. Rcan1 expression is preceded by Egr1 expression. Egr1 is a positive regulator of FcεRI-induced cytokine production. Biochemical and genetic evidence suggested that Egr1 controls Rcan1 expression. Our results identified Rcan1 as a novel inhibitory signal in FcεRI-induced mast cell activation and established a new link of Egr1 and Rcan1 in FcεRI signaling.
Lack of negative selection of CD4 T cells by bone marrow (BM)-derived cells causes fatal autoimmunity manifested as acute GVHD (141.16)
Authors: Chen Leo Cheng , Guo Xiaohua , Jensen Peter E , Chen Xinjian 
Published: January 2023

Abstract
While it is well established that BM-derived APCs play an essential role in the induction of self-tolerance, the biological consequence of losing this tolerogenic mechanism has yet to be determined. We generated BM chimeras by transplanting BM from MHC II-/- B6 mice into lethally irradiated Rag-/- or WT B6 mice. Lymphopoesis in the recipient mice initially appeared normal, but 5 wks post BMT, most mice became rapidly ill and died shortly afterwards. Histological examination revealed extensive epithelial apoptosis of the GI tract and skin on a pauci-inflammatory background. The liver showed marked vasculitis. The thymus underwent rapid atrophy and loss of developing thymocytes. The heart, lung, kidney, brain, salivary glands, pancreas, pancreatic islets and gonads were not affected. CD4 T cells in the thymus reacted vigorously against B6 DCs in vitro. Although thymic selection of CD8 T cells was unaltered, the CD8 T cells in the lymph nodes of the diseased animals were activated. Transfer of spleen cells from the diseased animals into Rag-/- mice resulted in similar tissue damage and accelerated death of recipients, but no disease if the Rag-/- mice were MHC II-/-. Further experiments involving MHC II-/- BM to MHC I-/- chimeras demonstrated that expression of MHC I by host tissue is also required for disease development. Our findings demonstrate that a consequence of lack of negative selection of CD4 T cells by BM-derived cells is fatal autoimmunity manifested distinctively as acute GVHD. The results also suggest that aGVHD can be induced solely by CD4 T cells generated in the recipients if the donor BM cells lack the MHC II type expressed by thymic epithelial cells of the recipients.
Rare dual TCR T cells contribute substantially to alloreactivity (141.2)
Authors: Morris Gerald P , Allen Paul M 
Published: January 2023

Abstract
T cell recognition of allogeneic MHC has long been recognized, though the nature of the alloreactive TCR repertoire is unknown. We hypothesize that the 1-3% of T cells expressing dual TCR due to incomplete allelic exclusion of TCRα are highly alloreactive due to non-thymically selected secondary TCR. Using the 4 available antibodies against TCRVα we observe an increased frequency of dual TCR T cells among alloreactive T cells in vitro. We quantitatively assessed the contribution of secondary TCR to the alloreactive repertoire by ELISPOT analysis; TCRA-C mice, incapable of expressing secondary TCRα, have a 50% decrease in alloreactive precursor frequency indicating secondary TCR contribute disproportionately to the alloreactive repertoire. We directly demonstrate expansion of the alloreactive repertoire by identification of a secondary non-thymically selected TCR with specific alloreactivity distinct from the primary TCR. The importance of dual TCR T cells is underscored by the dramatic increase of dual TCR among T cells mediating parent to irradiated F1 graft-vs.-host disease (GvHD); dual TCR comprised as much as 40% of peripheral T cells in mice with GvHD, compared to 3% observed in control mice. Together these studies convincingly demonstrate the importance of dual TCR T cells in alloreactivity. NIH AI061173 (PMA) and W.M. Keck Foundation Fellowship in Molecular Medicine (GPM).
The effect of the αGal epitope in the response to ECM in a nonhuman primate model (141.26)
Authors: Daly Kerry A , Stewart-Akers Ann A , Cordero Kevin , Johnson Scott A , Badylak Stephen F 
Published: January 2023

Abstract
The Galα1,3Galβ1,4GlcNAc-R (Gal) Epitope is a major factor in the hyperacute rejection of pig organs transplants in primates. Biologic scaffold materials used for tissue reconstruction and composed of xenogeneic extracellular matrix (ECM) may contain the Gal epitope. However, the effect this epitope has upon the host response is controversial. The present study investigated the effect of the Gal epitope upon the host response to a porcine-derived ECM in an African Green monkey (Cholrocaebus aethiops) abdominal wall resection model. Histologic methods, serology, complement-dependent cytotoxicity (CDC) and gene expression profiling were used to evaluate the host response to allogeneic and both wild-type and Gal-deficient xenogeneic scaffold materials. Although expression of the Gal epitope induced an increase in serum anti-Gal antibodies in recipients, no other differences were noted in the host response between test articles. All ECM scaffolds were well tolerated and showed constructive remodeling during the test period. Recipients of all test articles showed no histologic or humoral evidence of sensitization when a second scaffold was implanted 45 days after the original surgery. The findings of the present study show that the Gal epitope within porcine-derived ECM scaffold material causes a serum antibody response, but no adverse effect upon tissue remodeling.