bluuebunny/crossref_metadata_2025_split huggingface dataset:
Modeling the Maturation Process of Dendritic Cells (134.40)
Authors: Gonzalez Joanna , Moran Thomas M , Sealfon Stuart C 
Published: January 2023

Abstract
Dendritic cells are antigen-presenting cells responsible for the initiation of immunity. Immature DCs act as sentinels that, upon recognizing a pathogen, alter their gene expression pattern; the mature DCs then secrete signals to stimulate the appropriate immune response. One of these signals is the cytokine IL-12, composed of IL-12p35 and IL-12p40; the interaction of both subunits is required for the production of bioactive IL-12. A related cytokine, IL-23, is formed by the interaction of IL-12p40 with IL-23p19. The relative proportions of IL-23 and IL-12 depend on the pathogen recognized by the DC. Consequently, a model is being built in order to attain a more accurate understanding of IL-12 synthesis. The first part of the model encompasses the transcriptional activation of IL-12 genes. Previous research has identified some of the transcription factors involved and it has been found that the transcriptional regulation of IL-12 and IL-23 involves both common and distinct signaling pathways. However, further experiments are required to obtain a more complete signaling network from virus infection to the activation of these transcription factors. The second part of the IL-12 model involves the synthesis of bioactive IL-12 and mass-action kinetic equations have been developed to approximate the transcription and translation of the two subunits, their dimerization and secretion. Experimental data are currently being obtained for kinetic parameter estimation, as well as for testing the model; this proposed methodology can then be extended to other DC maturation signals in order to acquire a more complete understanding of the initiation of immunity.
Ibuprofen, the popular over-the counter non-steroidal anti-inflammatory drug, blunts human antibody synthesis in vitro (134.41)
Authors: Bancos Simona , Phipps Richard P. 
Published: January 2023

Abstract
Background: Non-steroidal anti-inflammatory drugs (NSAIDs) are effective in reducing fever, relieving pain and inhibiting inflammation. NSAIDs function through inhibition of cyclooxygenases 1 and 2 (Cox-1 and Cox-2). Cox-1 is ubiquitously expressed, while Cox-2 expression is increased during inflammation and in malignant cells. We recently showed that Cox-2 is up-regulated in normal human B lymphocytes after activation and is required for optimal antibody synthesis. We hypothesized that ibuprofen (dual Cox-1/Cox-2 inhibitor) blunts antibody synthesis in human peripheral blood mononuclear cells (PBMC) and B lymphocytes.
Methods: IgM and IgG production in anti-IgM + CpG 2395 stimulated human PBMC and B cells was detected by enzyme-linked immunosorbent assay (ELISA). B cell proliferation and viability in the presence of ibuprofen was calculated using mitochondrial dehydrogenase activity (MTT) and 3H-Thymidine incorporation assays.
Results: Pharmacological concentrations of ibuprofen significantly reduced IgM and IgG synthesis in human PBMC and B lymphocytes in a dose-dependent manner. Ibuprofen had only a modest effect on B cell viability, but significantly inhibited B cell proliferation.
Conclusion: Ibuprofen reduces optimal antibody synthesis in part, by decreasing B cell proliferation.
Research supported by DE011390, AI071064, ES01247 grants.
Immunomodulatory activity of synthetic innate defence regulators (IDRs) (134.45)
Authors: Easton Donna M , Ma Shuhua , Mookherjee Neeloffer , Hamill Pamela , Lynn David , Gardy Jennifer , Mullaly Sarah , Pilat Sandra , Waldbrook Matthew , Thorson Lisa , Falsafi Reza , Chikatamarla Avinash , Team InnateDB , Schofield Louis , Finlay Brett , Brinkman Fiona , Hancock Robert EW 
Published: January 2023

Abstract
Cationic host defence (antimicrobial) peptides, e.g. cathelicidin LL-37, have a variety of immunomodulatory activities that favour the safe resolution of infections. We have studied novel synthetic cationic innate defence regulator peptides that are not directly antimicrobial but are anti-infective in vivo, due to modulation of innate immunity. A range of peptides derived from the small bovine peptide bactenecin were screened for immunomodulatory activities in vitro; e.g. promotion of chemokine production and suppression of pro-inflammatory cytokines. Since innate immunity is complex, involving &gt;1,500 gene products, a systems biology approach was utilized to characterize peptide modulation of innate immunity, including analysis of receptors, signalling pathways, transcription factors and downstream genes. To permit visualization and bioinformatic analysis of complex events, an innate immunity database (www.innatedb.ca), a network visualization tool (Cerebral) and downstream analysis tools (e.g. pathway overrepresentation analysis) were developed and provided insight into how this selective modulation occurs. In vivo data indicate that these activities provide protection in animal model infections of Gram positive and Gram negative bacterial infections as well as severe malaria. Thus IDRs have great potential for use as novel anti-infective agents.
Supported by Genome BC, FNIH and CIHR.
Glycyrrhizin improves the antimicrobial peptide production by human epidermal keratinocytes (NHEK) transwell-cultured with peripheral blood linage- CD34+ cells, a suppressor cell on the antimicrobial peptide production by NHEK (134.46)
Authors: Yoshida Shohei , Jones Vickie C , Jeschke Marc G , Kobayashi Makiko , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
Previously, linage- CD34+ cells isolated form burn patient peripheral blood cells were shown to be inhibitory on the antimicrobial peptide production by NHEK. In the present study, the effect of glycyrrhizin (GL) on the CD34+ suppressor cell activity for the production of β-defensin 1 (BD-1) by NHEK was determined. Linage- CD34+ cells were isolated from peripheral blood of healthy donors after 1-hr of programmed exercise, as previously described. CD34+ cells (1 x 105 cells/ml, upper chamber) were transwell-cultured with NHEK (1 x 105 cells/ml, lower chamber) in the presence or absence of GL (Minophagen Pharmaceutical Co., Ltd., Tokyo, Japan). Forty-eight hrs after cultivation, cells in the lower chamber were assayed for human BD-1 and cathelicidin (CAP-18) mRNAs by RT-PCR. BD-1 and CAP-18 mRNAs were consistently expressed by NHEK. However, NHEK transwell-cultured with CD34+ cells did not express these peptide mRNAs. When transwell-cultures between these 2 cells were performed in the presence of 100 μg/ml of GL, mRNAs for BD-1 and CAP-18 were expressed by NHEK. The mRNA expression for BD-1 and CAP-18 by NHEK was not directly influenced by GL. These results indicate that the suppressor cell activity of linage- CD34+ cells on the antimicrobial peptide production by NHEK is modulated by GL. This study was supported by SHC NA #8610.
Improved survival in experimental sepsis with conjugated cholic acids preventing activation of dendritic cells by LPS (134.48)
Authors: Kim Youn-Hee , Seong Seung-Yong 
Published: January 2023

Abstract
The incidence and mortality of sepsis have steadily increased during the last few decades. The development of targeted therapy has been hampered due to multiple targets of LPS. Here, we tested bioemulsifiers inhibiting interaction of LPS with dendritic cells (DCs) in vitro and in an experimental mouse model. We found that glycine-conjugated cholic acids (CCA) and taurine-CCA reduced LPS-mediated NF-κB activation by inhibiting LPS binding with target cells. CCAs were also able to inhibit expression of costimulatory molecules, production of inflammatory cytokines and NO of DCs by LPS. In addition, CCAs inhibit T cell proliferation by DCs activated with LPS. In vivo, CCA administration reduced blood proinflammatory cytokines and improved mice survival following LPS challenge. Considering functional impairment of DCs in sepsis patients, protecting DCs from hyperactivation by LPS might be crucial in the physiological outcome of sepsis. Taken together, CCA might mitigate the development of severe sepsis.
The role of IL-1β in the vaccine adjuvant Monophosphoryl Lipid A’s TRIF-biased TLR4 signaling and low toxicity (134.49)
Authors: Eaves Chelsea Anne , Cekic Caglar , Mitchell Thomas C. 
Published: January 2023

Abstract
Vaccines often require adjuvants to elicit responses to otherwise weakly immunogenic proteins. The TLR4 agonist, monophosphoryl Lipid A (MLA) retains the immunogenicity, but not the toxic inflammatory response, of its parent molecule S. minnesota LPS. We previously reported that MLA is a TRIF-biased agonist of TLR4, and have now determined that a single phosphate group difference is sufficient for this signaling difference, as well as for a loss of IL-1β production. These two characteristics of MLA contribute greatly to its reduced toxicity. In order to determine the relationship between TRIF-biased signaling and the loss of IL-1β, we compared IL-1β production by RT-PCR and ELISA in primary bone marrow-derived dendritic cells. Preliminary studies show that MyD88 signaling is required for post-transcriptional IL-1β processing, indicating that TRIF-bias may cause MLA to produce low levels of IL-1β protein. Discovering the mechanisms responsible for these low-toxicity adjuvant properties of MLA will aid in the generation of safer and more effective vaccines. This project was supported by funding from the NIH/NIAID.
Rational design of enhanced affinity at the MICA-NKG2D interface (134.51)
Authors: Stoufer Lisa K. , Davies Luke D. , Culpepper David , McFarland Benjamin J. 
Published: January 2023

Abstract
MICA is an MHC-like protein expressed on stressed cells. It binds the NKG2D immunoreceptor and activates natural killer cells. Our research seeks to enhance binding affinity at contacts between MICA and NKG2D. Previous research has focused on mutating the area beneath the binding site, an indirect design strategy that has increased the association and affinity between MICA and NKG2D by more than an order of magnitude. We currently aim to increase binding affinity directly by designing point mutations at MICA-NKG2D protein-protein contacts. RosettaDesign was used to predict which contact mutations would be most beneficial at the MICA-NKG2D interface. We have produced multiple point mutants of MICA in an undergraduate research environment and measured NKG2D binding kinetics and thermodynamics using surface plasmon resonance. In the first round of design, 4 out of 6 mutations enhanced NKG2D affinity by 0.5 kcal/mol or more, up to 1.2 kcal/mol. This success rate is similar to published accounts of antibody-antigen design and other protein design strategies. Once a panel of point mutations is complete, we will investigate the kinetics and thermodynamics of combining mutations, from direct and indirect design strategies.
Meprin metalloproteases are involved in monocyte trans-extracellular matrix migration and meprin deficiency aggravates LPS-induced septic shock (134.56)
Authors: Jin Hong-Jian , Banerjee Sanjita , Yura Renee , Bao Lei , Bond Judith , Sun Qi 
Published: January 2023

Abstract
Meprins are metalloproteases first discovered in kidney and intestinal epithelial cells, and have been implicated in autoimmune diseases. We have shown that homeostasis of monocytes is compromised in meprin alpha/beta null (alpha-/-/beta-/-) mice. However, the mechanisms responsible for the involvement of meprins in the immune and hematological system remain uncharacterized. We report here that human monocytes expressed meprins, and the expression of meprins was down-regulated in mature macrophages and dendritic cells but not their immature counterparts. We further show that bone marrow monocytes and peritoneal macrophages from meprin-deficient mice had decreased efficiency in migration through extracellular matrix (ECM) compared to those from the wild-type mice in Boyden assays without marked defects in cell mobility in the absence of ECM. In a sepsis shock model, meprin-deficient mice exhibited more severe hypothermia than the wild-type mice in response to peritoneally injected lipopolysaccharide (LPS), and the prevalence of blood monocytes in the merpin-deficient mice were only 25% of that in the wild-type mice, with compensatory increase of lymphocytes. While meprin-deficient mice in the steady state had moderately lower blood levels of IL-12, MCP-1 and IL-6 than the wild-type mice, LPS-challenged meprin-deficient mice exhibited a drastic elevation of the above inflammatory cytokines in comparison to the wild-type mice, with comparable blood concentrations of IFN-gamma and IL-10 between the two mice. These results indicate that meprin metalloproteases are required for efficient trans-ECM migration of monocytes, and meprin deficiency is associated with defects in monocyte functions which have pathological consequences.
Heat shock protein 90 protects degradation of retinoic acid-inducible gene-I (134.75)
Authors: Matsumiya Tomoh , Imaizumi Tadaatsu , Yoshida Hidemi , Topham Matthew K. , Stafforini Diana M. , Satoh Kei 
Published: January 2023

Abstract
Retinoic acid-inducible gene I (RIG-I) plays a pivotal role in innate immune responses as an intracellular sensor of viral RNAs. Recognition of dsRNA by RIG-I activates downstream IFN-inducible molecules including IRF3, resulting in the induction of type I IFN. However, the mechanisms that participate in downstream events following activation of RIG-I are incompletely understood. The objective of this study was to identify proteins that associated with RIG-I. Studies combining immunoprecipitation and mass spectrometry showed that RIG-I formed part of a protein complex that included heat shock protein 90 (HSP90), a molecular chaperone. Treatment with geldanamycin, a HSP90 inhibitor, dissociated the RIG-I: HSP90 complex and also resulted in the degradation of RIG-I protein. In contrast, the levels of RIG-I mRNA were not affected. In addition, the proteasomal inhibitor MG-132 significantly abolished geldanamycin-mediated RIG-I degradation, suggesting that HSP90 protected RIG-I from proteasomal degradation. Our results also showed that pretreatment with geldanamycin significantly reduced RIG-I mediated IFN-β promoter activity induced by polyinosinic: polycytidylic acid. Taken together, these data suggest that HSP90 plays a role in innate immune responses by shielding RIG-I from proteasomal degradation.
STAT6 Independent Alternative Activation of the Pulmonary Environment in Response to Helminth Infection. (134.78)
Authors: Siracusa Mark  C. , Craig John M. , Urban Joseph F , Artis David , Pearce Edward J 
Published: January 2023

Abstract
Macrophages adopt an alternatively activated phenotype in the context of a polarized Th2 response as a result of STAT6 (Signal transducer and activation of transcription 6)-dependent IL-4 and IL-13 signaling. Given the strong connection between IL-4/IL-13/STAT6 signaling and the induction of the alternatively activated macrophage phenotype, it was unexpected to observe that infection with Nippostrongylus brasiliensis (Nb), a strong inducer of IL-4 and IL-13, in Stat6-deficient mice resulted in an apparent alternate activation of lung macrophages with the induction of arg1, ym1 and fizz1. The goal of the work presented here was to test the hypothesis that there is a STAT6-independent pathway for alternative activation of lung macrophages in response to the pulmonary inflammation induced by the transient passage of Nb larvae through the parenchyma of the lungs. To test this hypothesis, we isolated CD11c+ lung macrophages from STAT6-/- mice on days 0, 2, 4, 8, and 12 post-Nb to test for the alternative activation. We demonstrate the macrophage-specific induction of Arg1 and to a lesser extent YM1 protein in an IL-4 and IL-13 independent manner. It was also determined that epithelial cells, but not macrophages, produce Fizz1 in a Stat6-independent manner. These results demonstrated the partial alternative activation of macrophages in the absence of IL-4 and IL-13 signaling and underscore the need to study the innate immune mechanisms capable of altering macrophage phenotypes.
Select procyanidins prime the γδ T cell by modulating transcriptome degradation (134.88)
Authors: Holderness Jeff S , Daughenbaugh Katie , Kimmel Emily , Freedman Brett , Hedges Jodi , Jackiw Larissa , Jutila Mark A 
Published: January 2023

Abstract
Tannins from many plant-derived traditional medicines demonstrate immune regulatory activity. Recently, we reported that condensed tannins (procyanidins) isolated from a select number of traditional medicines act as γδ T cell agonists. γδ T cells play an important role in mucosal immunity by regulating the mucosal immune response and maintaining epithelial health. Procyanidins induce a priming state on the γδ T cell, thus improving its ability to quickly respond to secondary stimuli. This priming state is characterized, in part, by an increased half-life of select transcripts in the γδ T cell relevant to innate immunity. This increase in transcript half-life could enable the cell to more quickly produce appropriate immune responses to secondary stimuli. Previously we reported that oligomeric procyanidin fractions, but not monomeric procyanidins, act as γδ T cell agonists. We have further defined the activity of different sizes of procyanidins. The tetrameric and pentameric procyanidins from apple peel have greater specificity for γδ T cells, whereas larger oligomers demonstrate less specificity. Our results suggest procyanidins with specific oligomeric structures prime γδ T cells by stabilizing transcripts with critical functions in innate immunity. Funded by: NIH/ NIAID Contract HHSN266200400009/N01-AI40009 and NCCAM P01 AT004986-01
Divergent macrophage programs for phagocytosis and antimicrobial responses regulate innate immunity to microbial pathogens (134.90)
Authors: Montoya Dennis , Cruz Daniel , Zhang Xiaoran , Krutzik Stephan , Ferguson Benjamin G , Modlin Robert L 
Published: January 2023

Abstract
The mechanisms by which monocytes differentiate into macrophages (Mø) contribute to the outcome of the host response to microbial infection. Previously, we found that IL-10 derived Mø (CD209+ M2) had enhanced phagocytic receptor expression and function versus IL-15 derived Mø (CD209+ M1). Here we show that CD209+ M2 Mø unlike CD209+ M1 Mø, were unresponsive to vitamin D for the induction of the antimicrobial peptide, cathelicidin, known to kill intracellular mycobacteria, demonstrating a divergence of phagocytosis and host defense function.. The divergent Mø programs for phagocytosis and antimicrobial responses regulate the effector arm of innate immunity relative to pathogenesis in microbial infection.
Receptor Like Tyrosine Phosphatases CD45 and CD148 Regulate Integrin- and GPCR-Mediated Pathways in Murine Neutrophils (135.18)
Authors: Zhu jing W , Doan Kristen , Zhang Hong , Lowell Cliff , Weiss Arthur 
Published: January 2023

Abstract
Reciprocal regulation of tyrosine phosphorylation by protein tyrosine kinases and phosphatases is central to normal immune cell function. CD45 is a very abundant and important receptor like tyrosine phosphatase in the immune system that regulates key proximal signaling molecules, Src family kinases. Interestingly, in contrast to its requirement in T cell development, the phenotype of CD45 deficient B cells and myeloid cells is rather subtle. Using a doubly deficient CD45 and CD148 mouse model, we discovered the overlapping function of these structurally distinct phosphatases in regulating immunoreceptor signaling via dephosphorylation of the inhibitory tyrosine of Src-family kinases in B cells and macrophages.
Since SFKs are also critical regulators of many other signaling pathways, including integrin and GPCR signaling, we studied the functions of CD148 and CD45 in these pathways. We focused on integrin- and GPCR-mediated pathways in neutrophils. We found that the superoxide release in response to integrin stimulation was impaired in both CD45 and CD148 deficient neutrophils and even more so in doubly deficient cells. Similar defects were observed when cells were stimulated with the GPCR ligand fMLP. Interestingly, we also observed impairment of calcium flux in CD148 single deficient neutrophils following MIP-2 stimulation, indicating unique function of CD148 in response to a chemokine signal. More detailed investigations on the biochemical and functional basis of these defects are under way.
This work is supported by NIH grant A1066120
Conditional targeting of C-terminal Src kinase (Csk) in NK cells (135.2)
Authors: Hampshire Daniel , Saudemont Aurore , Tarakhovsky Alexander , Colucci Francesco 
Published: January 2023

Abstract
Csk is a key negative regulator of Src-family kinases and has been shown to be a strong inhibitor of immunoreceptor signalling in T cells and granulocytes. Our hypothesis is that an increase in Src kinase activity might take place in NK cells following the elimination of Csk, resulting in an enhanced cellular response even at low levels of stimulation. We aim to establish the role of Csk in NK cells using conditional knockout mice. A transgenic mouse has been developed where the Csk gene is flanked by LoxP (floxed) sites. Crossing Csk floxed transgenic mice with transgenic mice expressing Cre recombinase under the control of a human CD2 gene (hCD2-iCre) will enable the deletion of the Csk gene in a tissue-specific manner. The ROSA26 reporter gene system suggests that Cre deletes the floxed gene in approximately 30% of splenic NK cells and, as reported before, in the vast majority of T cells and B cells. Intracellular staining for Csk demonstrates a correlation between Cre targeting of the ROSA26 locus and the target locus. There is a reduction in the number of lymphocytes in the Csk knockout (KO) mouse. More specifically, there is a decrease in B cells, suggesting a role for Csk in B cell development. However, no obvious phenotype is detected in NK cells. Ongoing studies of NK cell functions will be presented.
Fc-receptor crosslinking stimulates cell cycle progression of macrophages via the ERK pathway (135.25)
Authors: Casadevall Arturo , Pollard Jeffrey W. , Luo Yong 
Published: January 2023

Abstract
Our previous study showed that phagocytosis and Fc receptors (FcR) crosslinking of macrophages could enhance their cell proliferation. In the present study, we further explored the activation of the signaling pathways responsible for this phenomenon. Our investigation revealed an increase in cyclin D1 and cyclin E protein expression in murine macrophages after FcR crosslinking. Specifically, we further identified the extracellular signal-regulated kinase (ERK) signaling pathway as a potential mediator of signals from FcR activation to cyclin D1 expression, since cyclin D1 expression associated with FcR crosslinking was attenuated by specific inhibitors of the ERK1/2 signaling pathway, PD98059 and U0126 and the spleen tyrosine kinase (Syk) inhibitor, Piceatannol. Our findings establish a link between the ERK and cyclin signaling pathways thus providing a mechanism by which FcR activation produces a mitogenic effect that stimulate macrophage proliferation. The results suggest the possibility of new treatment options for certain infectious diseases, chronic inflammatory diseases and leukemias based on interference with FcR-mediated cell cycle progression.
This work was supported by NIH grants AI033142, AI033774, AI052733, HL059842, 5T32CA009173, 5R01HD030280-09, 5RO1CA089617-03 and the Northeastern Biodefense Center grant AI057158-05.