bluuebunny/crossref_metadata_2025_split huggingface dataset:
Innate Defence Regulator Peptides as a Novel Anti-infective Strategy (94.2)
Authors: Ma Shuhua , Nijnik Anastasia , Madera Laurence , Elliott Melissa , Easton Donna , Mayer Matthew , Kindrachuk Jason , Jenssen Håvard , Mok Ka Yan (Winnie) , Yang Tom , Hancock R.E.W. (Bob) 
Published: January 2023

Abstract
The therapy of bacterial infections is under great threat as multiple antibiotic resistance increases and there is a paucity of new antibiotic discovery and development. Synthetic Innate Defence Regulator (IDR) peptides, which mimic natural host defence (antimicrobial) peptides, have been designed as a novel anti-infective strategy, working by selectively boosting innate immune protective mechanisms while dampening potentially harmful inflammation. These peptides can resolve serious infections in animal models. The optimal method of use of these peptides was evaluated here using an animal model infection by one of the highly resistant bacteria (Superbugs) afflicting our society, Staphylococcus aureus. A luminescent version of this bacterium was utilized to follow the kinetics of infection non-invasively using IVIS imaging. Protection was achieved by both prophylactic and therapeutic administration. Investigation of the mechanism of protection by examining cells infiltrating the infection site and cytokines/chemokines revealed remarkable parallels between in vitro action of the peptides in primary human cells and these animal models. System biology approaches such as Microarray and InnateDB are being utilized to decipher the complex. This has revealed several receptors, signaling pathways, transcription factors and effector proteins involved in the modulation by peptides of innate immunity.
Critical roles of NKp30 and NKG2D in synapse formation and activation of NK cells (94.8)
Authors: Zheng Xiaodong 
Published: January 2023

Abstract
Natural killer (NK) cell is one of innate immune cells with potential of killing target cell without pre-activation. Activating killer receptors NKp30 and NKG2D play important roles in NK cytotoxicity, however, the critical roles of NKp30 and NKG2D in NK cell immunological synapse remains incompletely understood. In this study, the cytotoxicity of NK92 cell against Hela cell with or without blocking NKp30 or NKG2D was measured. The NK92-Hela conjugates were performed by immunofluorescence and Flow cytometry. Blockade of NKp30 or NKG2D inhibited NK cytotoxicity against several MHC I deficient targets. The cytotoxicity of NK92 cells against Hela was lower with blocking NKp30 than that with blocking NKG2D. The same effect of blocking NKp30 or NKG2D was detected in fresh NK cells from PBMC. However, blockade of NKp30 and NKG2D could not decrease the binding rate of NK92 and Hela cells. These results suggest that NKp30 and NKG2D involve in activation but not complete combining with target cells in NK synapse formation.
Human CD1D gene expression is regulated by LEF-1 through distal promoter regulatory elements (95.4)
Authors: Chen Qiaoyi , Zhang Tao , Pincus Seth , Wu Shixiu , Ricks David , Liu Donald , Sun Zhongsheng , Maclaren Noel , Lan Michael 
Published: January 2023

Abstract
CD1d expressing cells present lipid antigen to CD1d-restricted natural killer T (NKT) cells, which play an important role in immune regulation and tumor rejection. Lymphoid enhancer-binding factor-1 (LEF-1) is one of the regulators of the Wnt signaling pathway, which is a powerful regulator in cellular growth, differentiation, and transformation. There is little evidence connecting Wnt signaling to CD1d expression. In this study, we have identified LEF-1 as a regulator of the expression of the gene encoding the human CD1d molecule (CD1D). We found that LEF-1 binds specifically to the CD1D promoter. Over-expression of LEF-1 in K562 or Jurkat cells suppresses CD1D promoter activity and down regulates endogenous CD1D transcripts, whereas knockdown of LEF-1 using LEF-1-specific siRNA increases CD1D transcripts in K562 and Jurkat cells but different levels of surface CD1d on these two cell types. Chromatin immunoprecipitation showed that the endogenous LEF-1 factor is situated at the CD1D promoter and interacts with histone deacetylase-1 to facilitate the transcriptional repressor activity. Knockdown of LEF-1 using siRNA potentiates an acetylation state of histone H3/H4, supporting that LEF-1 acts as a transcriptional repressor for CD1D gene. Our finding links LEF-1 to CD1D and suggests a role of Wnt signaling in the regulation of human CD1D gene.
IL-23 receptor deficiency prevents the development of lupus nephritis in B6/lpr mice (96.20)
Authors: Zhang Zheng , Kyttaris Vasilieios , Kuchroo Vijay , Oukka Mohamed , Tsokos George 
Published: January 2023

Abstract
IL-17 producing T cells infiltrate kidneys of patients with lupus nephritis, and IL-23-treated lymphocytes from lupus-prone mice may transfer disease to Rag-1-/- mice. Here we show that IL-23 receptor deficient lupus prone B6/lpr mice display decreased numbers of CD3+CD4-CD8- cells and IL-17 producing cells in the lymph nodes, and produce less total serum IgG and anti-double strand DNA antibodies. In addition, clinical and pathology measures of lupus nephritis are abrogated. The presented experiments imply the importance of IL-23 receptor medicated signaling in the development of lupus nephritis and possible drug target for the treatment of the disease.
APPLICATIONS OF IMMUNOSUPPRESSIVE DNA TO TREAT AUTOIMMUNE AND AUTOINFLAMMATORY DISEASES (96.22)
Authors: Yagci Fuat , Aslan Ozlem , Songur Murat , Karatepe Kutay , Gursel Ihsan 
Published: January 2023

Abstract
Synthetic oligodeoxynucleotides (ODN) expressing suppressive TTAGGG motifs effectively down-regulate the production of proinflammatory and Th1 cytokines elicited either by Toll-Like Receptor (TLR) dependent or independent immune stimuli. Although initially identified by their ability to block CpG-induced immune activation, it was shown that suppressive ODN was able to block multiple forms of immune stimulation and to be effective in the prevention and treatment of pathologic autoimmune and autoinflammatory diseases. Endotoxin-induced uveitis (EIU) is an established animal model of acute ocular inflammation. It is induced by either systemic or intravitreal administration of lipopolysaccharide (LPS). Familial Mediaterranean Fever (FMF) is an autosomal recessive periodic fever disease characterized by recurrent, self-limiting, febrile, inflammatory attacks of the serosal membranes such as peritoneum, pleura, and synovia. Present study aims to demonstrate the downregulatory effect of the suppressive DNA on EIU and FMF as local autoimmune and systemic autoinflammatory diseases respectively. Results indicated that LPS induced EIU in mice, A151 treatment strongly downregulated either expression or protein levels of IL6, IP10, IL1β, MIP1β, MIP3a or iNOS. Moreover, FMF patient PBMCs were more responsive to TLR ligand stimulation compared to healthy subjects and A151 strongly reversed this activation and suppressed several key Th1-cytokine/chemokine levels
Small Modular ImmunoPharmaceutical (SMIP™) Molecules Directed at the TCR Complex (CD3) Block T Cell Activation and Cause Minimal Cytokine Release In Vitro (96.25)
Authors: Wang Camilla , Beckett Travis , Odegard Valerie , Hussell Scott , Mohler Ken , McMahan Catherine 
Published: January 2023

Abstract
SMIP molecules are single chain binding proteins based on a scFv-CH2-CH3 format. We have generated SMIP proteins using binding domains from well characterized mitogenic and non-mitogenic anti-CD3 antibodies, and have found dramatic differences in the biologic consequences on human T cells exposed to the compounds. In particular, compared to antibodies, SMIP proteins cause a unique Ca++ flux, are better able to down modulate the TCR complex from the cell surface, and can inhibit naïve and activated T cells in various in vitro assays. Importantly, SMIP proteins cause a much reduced level of cytokine release, notably, when compared to second generation anti-CD3 antibodies. Most known anti-CD3 antibodies cause activation of T cells, especially when cross linked via FcR on the surface of APCs. Without cross linking, these antibodies have minimal activation of T cells in vitro. In the clinic, second generation anti-CD3 antibodies are still plagued with cytokine release syndrome, despite showing little detectable cytokine release from primary T cells in vitro. We have developed an activated T cell blast assay sensitive enough to detect in vitro cytokine release caused by anti-CD3 antibodies in second generation formats. Results from these assays indicate that features in the SMIP format could generate novel CD3-directed therapeutics with superior efficacy and reduced potential for the serious side effects of Cytokine Storm.
Delta-like ligand 4 regulation of effector T cell function during experimental autoimmune encephalomyelitis (96.8)
Authors: Reynolds Nathan , Lukacs Nicholas , Karpus William 
Published: January 2023

Abstract
Notch signaling mediates cell fate decisions through specific ligand-receptor binding. Signaling by Delta-like ligand 4 (DLL4) influences both CD4+ Th1 and Th17-associated cytokine production and contributes to adaptive immunity. Experimental autoimmune encephalomyelitis (EAE) is an autoimmune disease of the central nervous system (CNS) that is mediated by autoreactive Th1 and Th17 CD4+ T cells. We examined the contributions of DLL4 in regulating Th1 and Th17 functions during EAE. CNS-infiltrating myeloid dendritic cells, macrophage and resident microglia express high levels of DLL4 during EAE. Blocking DLL4 during antigen-restimulation of primed T cells resulted in decreased inflammatory cytokine production by these T cells. Administration of anti-DLL4 to PLP139-151-primed mice decreased EAE severity. Similarly, pretreatment of PLP139-151-specific T cells with anti-DLL4 reduced adoptive disease transfer. Treatment of mice with anti-DLL4 following adoptive transfer of primed T lymphocytes blocks localization of these T cells to the CNS and prevents disease. These results suggest DLL4-Notch signaling regulates autoimmune T cell effector function and localization to the CNS. (Supported by NMSS RG4146-A-4)
Tannic acid and quercetin display a therapeutic effect on atopic dermatitis via suppression of angiogenesis and Th2 polarization. (97.7)
Authors: Jung Min Kyung , Cho Dae-Ho 
Published: January 2023

Abstract
Atopic dermatitis (AD) is a chronic inflammatory skin disease that is characterized by inflammation, abnormal angiogenesis and elevated Th2 polarization. Tannic acid (TA) and quercetin are well known polypenolic compounds, have documented anti-inflammatory effect. However, little is known about the effects of TA and quercetin on atopic dermatitis and related mechanisms. The present study investigated whether TA and quercetin had a therapeutic effect on atopic dermatitis. In vitro, we found that TA and quercetins reduced pro-angiogenic factor and Th2-inducing cytokine in human keratinocytes. The inhibitory effect was higher in the combined application of the two agents than in the application of one agent alone. To investigate the effect of combined application with TA and quercetin in vivo, an AD-induced Nc/Nga mouse model was used. The group receiving TA and quercetin showed disease improvement with down-regulation of serum IgE, TARC production and angiogenesis level compare to non-treated group. Taken together, these data demonstrate that TA and quercetin have a therapeutic effect on AD by suppressing angiogenesis, Th2 polarization.
Dendritic Cell subsets: Functional analysis and TLR Phenotyping (98.13)
Authors: Rosenberg Jonathan , Chitta Sriram , Singh Gita , Quintel Payton , Lee Hyun-Ku , Singh Sujay 
Published: January 2023

Abstract
In-depth analysis of subtypes of Dendritic Cells (DCs), both myeloid (mDC) and plasmacytoid (pDC) subsets, has been developed through Toll-like Receptor (TLR) Phenotyping by Flow Cytometry. Sets of TLR specific mAbs are combined with cell lineage and DC markers like CD11c and CD123 for mDC and pDC, in a simple assay for consistent DC analysis. DC subsets in peripheral blood differ in origin, phenotype, and function as analyzed by cytokines, chemokines or pro-inflammatory factors produced upon interaction with pathogens. DC response to invading pathogens depends upon expression of a spectrum of receptors, such as TLRs, which are expressed on the cell surface and in intracellular endosomes. TLR signaling in DC subtypes results in production of cytokines, chemokines and pro-inflammatory factors that elicit and control adaptive immune responses. TLR Phenotyping analysis includes development of 4 color Flow Cytometry assay cocktails for TLR expression of DCs to characterize mDC or pDC in human peripheral blood. In combination with activation and functional markers such as cytokines or chemokines the assay system provides a new research tool to identify changes in TLR expression in steady state or pathogen infected conditions and during differentiation of monocytes, DCs, B cells/B cell lines (EBV transformation), or T cells. The approach also provides additional information on TLR signaling and downstream pro-inflammatory mediator production.
LAB/NTAL regulates zymosan-mediated inflammatory responses in Bone Marrow Derived Dendritic Cells (BMDC) (98.14)
Authors: Orr Selinda , Quigley Laura , Roessler Stephen , Hodge Deborah , Razzook Catherine , Cardone Marco , Zhang Weiguo , Trinchieri Giorgio , Lyakh Lyudmila , McVicar Daniel 
Published: January 2023

Abstract
Immune sensing of fungi occurs via multiple pathogen recognition receptors such as Dectin-1/2, mannose receptor and TLR2. The β-glucans and mannans in fungal cell walls activate these receptors. Stimulation of DCs via TLR2 results in the production of IL-10 and little IL-12, while Dectin-1 utilizes a hemi-ITAM (immunoreceptor tyrosine based activation motif) and Syk to facilitate IL-23 production leading to Th17 polarization. We have recently shown that the linker for activation of B cells/Non-T cell activating linker (LAB/NTAL), a known regulator of ITAM-mediated signaling, is expressed in BMDCs. Here we examine whether LAB regulates the response of BMDC to zymosan by affecting Dectin-1 signaling. We find that LAB, Syk, Cbl, MAPK and Akt are rapidly and transiently phosphorylated following zymosan stimulation. Moreover, zymosan stimulated Lat2-/- BMDC produced decreased IL-10 and IL-12p40 and increased TNF levels suggesting the involvement of LAB in regulating BMDC cytokine production. Unexpectedly, direct engagement of Dectin-1 by purified β-glucans failed to elicit LAB phosphorylation. Interestingly, water-soluble components of zymosan induced phosphorylation of FcRγ, Syk, and LAB. Indeed, purified yeast-derived mannans also stimulated phosphorylation of Syk and LAB in BMDC. These data indicate that LAB regulates the BMDC inflammatory response to fungal pathogens not by affecting Dectin-1 signaling but by regulating mannan-derived signals likely through Dectin-2.
The role of surface charge in cationic liposome-induced antigen-presenting cell (APC) activation and viability (98.21)
Authors: Ma Yifan , Xie Xiaofang , Zhuang Yan , Zhou Dongmei , Cai Lintao 
Published: January 2023

Abstract
Cationic liposomes, an effective carrier for drugs and gene delivery, have recently been reported as a potential vaccine adjuvant. Since the immuno-stimulatory effect of cationic liposomes may be related to their surface charge, we hereby investigated the effect of liposome charge on antigen-presenting cell (APC) activation and viability. The charge of cationic liposomes was adjusted by incorporating into DOTAP (a cationic liposome) variable amounts of DOPC (a neutral lipid). DOPC by itself did not affect monocyte activation or DC maturation. The DOTAP/DOPC liposomes with molar ratios of 4:1, 3:2 or 2:3 not only elevated CD86 on monocytes and induced the production TNF-alpha, but also significantly augmented the expression of CD83 and CD86 on both human and mouse dendritic cells (DCs). However, further increasing the proportion of DOPC significantly attenuated monocyte activation and DC maturation induced by DOTAP/DOPC liposomes. Interestingly, although DOTAP had a higher zeta potential than the DOTAP/DOPC liposome (4:1), it was less potent in inducing monocyte activation and TNF-alpha production. Moreover, the treatment of monocytes with DOTAP for 24 hr caused about 40% apoptosis, while the presence of DOPC significantly increased monocyte viability. Hence, an appropriate surface charge is crucial for enhancing the adjuvant effect of cationic liposomes and reducing their potential cytotoxicity.
Molecular mechanisms of stress-induced glucocorticoid resistance in splenic macrophages: A role for p38 MAPK (98.22)
Authors: Stiner LaTonia , Sheridan John 
Published: January 2023

Abstract
Stress has been shown to increase production of glucocorticoids (GC’s), which suppress the inflammatory response. However, the increased levels of circulating GC’s observed following social disruption stress (SDR) fail to suppress cytokine production or induce apoptosis in splenic CD11b+ cells from WT mice, suggesting that these cells are GC resistant. Moreover, CD11b+ cells from IL-1R1 knockout (KO) mice are not GC resistant following SDR. The results of the present study show that following LPS and corticosterone (LPS/cort) stimulation, CD11b+ cells from WT SDR mice displayed an increase in cytoplasmic p38 phosphorylation (p-p38) when compared to controls. The increase in p-p38 was associated with an increase in nuclear translocation of the GC receptor phosphorylated at Ser234 (GR-pS234), within SDR mice compared to controls. Studies conducted with p38 and JNK inhibitors revealed that a reduction in p38 activity led to an increase in JNK activation and that cytoplasmic GR-pS234 was maintained in SDR and control mice. Furthermore, p-p38 was notably absent in KO macrophages from SDR and control mice stimulated with LPS/cort. These data confirm that SDR enhances GR-pS234 and JNK may facilitate this phosphorylation in the absence of p-p38. These data also support a role for p38 in induction of GC resistance.
Expansion, mobilization, and circulation of myeloid derived cells in mammary tumor bearing (TB) mice. (98.27)
Authors: Talmadge James , Younos Ibrahim , Dafferner Alicia , Samson Holly , Westphal Sherry , Fuminori Abe 
Published: January 2023

Abstract
MDSCs are increased in the marrow (BM), lungs, liver, and tumors of TB hosts as compared to non-TB (NTB) mice. In vivo, BrdU labeling of leukocytes from 4T1 mammary TB mice revealed that MDSCs had an increased rate of proliferation in BM, spleen, lungs, and liver. MDSCs are observed in the highest frequency in the blood and spleen, suggesting hematopoietic progenitors mobilized from BM and proliferated following arrest. We assessed the biodistribution of circulating cells using CFSE-labeled spleen cells from TB mice, adoptively transferred to normal mice by iv injection and rapidly arrested in the lungs. In contrast, CFSE-labeled MDSCs injected into 4T1 TB mice transversed the lungs, briefly arrested in the liver, and were retained in BM and tumors. We posit that in 4T1 TB mice, the normal arrest in the pulmonary capillary bed is prevented by increased levels of nitric oxide from activated MDSCs resulting in capillary vasodilation and reduced arrest. In 4T1 TB mice, CD3+ T-cells are decreased, although the MFI of CFSE suggests proliferation. Annexin V staining revealed a high frequency of apoptotic CD3+ T-cells, associated with MDSC accumulation. In summary, MDSCs proliferate in lymphoid and parenchymal organs resulting in T-cell apoptosis possibly associated with activation-induced cell death. Further, although MDSCs proliferate in many organs, they rapidly circulate preferentially homing to the spleen and liver.
SHP-1 is a central regulator of dendritic cell function (98.3)
Authors: Ramachandran Indu , Song Weitao , Levitt Jonathan 
Published: January 2023

Abstract
Dendritic cells (DCs) are specialized antigen presenting cells that initiate T cell responses upon activation by pathogen. Once the pathogen is cleared, DCs regulate excessive T cell responses by turning on intrinsic inhibitory signals, the mechanisms of which are not entirely understood. SHP-1 is an inhibitory phosphatase that can deactivate signaling from a wide range of activating receptors such as antigen, cytokine and chemokine receptors in lymphocytes and macrophages. Despite our vast knowledge of the role of SHP-1 in several immune cell types, the specific role of SHP-1 in DCs remains unclear. To address the role of SHP-1 in DCs, we modulated SHP-1 activity in primary mouse bone-marrow DCs by using RNAi, chemical inhibition and SHP-1 functional mutants. We showed that in DCs, SHP-1 inhibited MAPK, NFκB, and AKT activity and lymph node (LN)-chemotactic receptor CCR7 expression, but had no effect on DC maturation. Consequently, SHP-1 inhibition lead to enhanced DC proinflammatory cytokine production, survival and LN migration. In vivo, vaccination with SHP-1-inhibited DCs lead to the expansion of tumor-specific cytotoxic T cells and reduction in FOXP3+ regulatory T cells resulting in effective anti-tumor immunity against mouse melanoma and prostate tumors. Collectively, these data demonstrate that SHP-1 functions as a central inhibitor of DC signaling and DC-induced T cell responses.
Establishment of central tolerance against blood-borne antigens by thymic Sirpα+ conventional dendritic cells (98.5)
Authors: Baba Tomohisa , Mukaida Naofumi 
Published: January 2023

Abstract
Intrathymic negative selection and differentiation of regulatory T (Treg) cells are a cardinal event in central tolerance system. Thymic dendritic cells (DCs) are presumed to be capable of inducing the apoptosis of autoreactive T progenitor cells and of alternatively differentiating them into Treg cells. In spite of the presence of heterogenous DC subsets in thymus, similarly observed on other lymphoid organs such as lymph nodes and spleen, the role of each intrathymic DC subset in the central tolerance is still unclear. Recently, we demonstrated that mice deficient in a CC chemokine receptor, CCR2, exhibited a selective diminution in CD11c+CD11b+CD8-Sirpα+ conventional DC (cDC) subset in the thymus with an accumulation of T cells with reactivity against serum self-antigen in the periphery. Here, we further revealed that Sirpα+ cDCs in the thymus can selectively capture blood-borne antigens due to their unique intrathymic localization nearby small vessels and inside perivascular regions, and then induce the antigen-specific Treg cells and subsequent clonal deletion to an intravenously injected antigen, depending on the intensity of antigen presentation. Finally, we established a novel way to in vivo expand antigen-specific Treg cells via intrathymic antigen presentation by thymic Sirpα+ cDCs, which can dampen aberrant immune reaction in an antigen-specific manner.