bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of chromatin remodeling during thymocyte differentiation (88.5)
Authors: Dovat Sinisa , Song Chunhua , Gurel Zafer , Payne Kimberly , Popescu Marcela 
Published: January 2023

Abstract
The Ikaros gene encodes a DNA-binding protein that functions as a master regulator of lymphoid development. Ikaros protein interacts with histone deacetylation complex and represses transcription of its target genes via chromatin remodeling. We have previously reported that Ikaros’ activity and protein stability is regulated by CK2 kinase (Popescu et al. J Biol Chem 2009 284:13869). We studied the effect of CK2-mediated phosphorylation on Ikaros function in primary thymocytes. Ikaros proteins with phosphomimetic mutations at CK2 phosphorylation sites 1) lose the ability to associate with Sin3a, a component of the NuRD histone deacetylase, as indicated by co-immunoprecipitation assays and 2) fail to repress genes involved in thymocyte differentiation as indicated by luciferase reporter assay. The introduction of phosphoresistant mutations at five N-terminal CK2 phosphorylation sites on the Ikaros protein restored Ikaros’ association with Sin3a, as well as wild-type levels of repressor activity. The treatment of primary thymocytes with CK2 kinase inhibitors resulted in increased levels of Ikaros protein and increased Ikaros’ DNA-binding affinity to the promoter regions of its target genes, as measured by quantitative chromatin immunoprecipitation. These results demonstrate that CK2 kinase regulates thymocyte differentiation by controlling Ikaros’ association with chromatin remodeling complexes and its ability to repress the transcription of developmentally regulated genes.
Conversion of Adenines to Inosines in RNA During Virus Infections Is an Innate Immune Stimulus (89.10)
Authors: Liao Jie-Ying , Prater Chrissy , Thakur Sheetal , Zalinger Zachary , Imani Farhad 
Published: January 2023

Abstract
Inflammatory responses that are generated during viral infections are critical for protective anti-viral immune responses. The exact viral-derived structures (pathogen-associated molecular patterns, PAMPs) that activate uninfected cells to induce pro-inflammatory signals are not completely characterized. To date, the viral PAMPs that are identified include double-stranded RNA (dsRNA) and 5'-triphosphate-containing single-stranded RNA (ssRNA). During viral infections, the cellular enzyme adenosine deaminase acting on RNA-1 (ADAR-1) catalyzes conversion of adenines to inosines, which is a pathway for blocking viral replication. ADAR-1 activation in the infected cells leads to the presence of inosine-containing viral and cellular RNA (Ino-RNA). Here, we report that Ino-RNA from infected cells is a stimulus for inflammatory innate immune response. Treatment of primary human bronchial epithelial (PHBE) cells, primary human alveolar macrophages or C57BL/J6 mice with in vitro synthesized Ino-RNA rapidly and potently induced several markers of innate immune response such as IFN-β, TNF-α, IL-6 and RANTES. We further identified increased secondary structures in ssRNA upon conversion of adenines to inosines. The increased secondary structures activated the dsRNA-specific binding protein PKR. In addition, Ino-RNA activated the MAP Kinase pathways, which is downstream of PKR. Our results suggest that Ino-RNA is a novel viral innate immune recognition element.
IL1 is Required for CD8 T Cell Stimulation of Endothelial Cells to Produce KC in Contact Hypersensitivity (89.13)
Authors: Kish Danielle , Gorbachev Anton , Fairchild Robert 
Published: January 2023

Abstract
Contact hypersensitivity (CHS) is a CD8 T cell-mediated response to hapten sensitization and challenge of the skin. Elicitation of CHS is initiated when hapten-primed CD8 T cells producing IFN-γ and IL-17 traffic to the challenge site and stimulate endothelial cells (EC) to produce CXCL1/KC. KC is required for the neutrophil recruitment directing CD8 T cell infiltration through the EC where they are activated to induce the edema of CHS. This study investigated the role of IL1 in the CD8 T cell priming and elicitation of CHS. Both KC production and neutrophil infiltration into the skin challenge site as well as CHS responses were low-absent following challenge of sensitized IL1R-/- vs. wild-type (WT) mice. Sensitization of IL1R-/- mice induced marked decreases in migration of hapten-presenting Langerhans cells into the skin-draining lymph nodes and priming of CD8+ T cells producing IFN-γ or IL-17 vs. WT mice. Transfer of WT hapten-presenting Langerhans cells to IL1R-/- mice did not correct these defects. Transfer of primed WT CD8 T cells to naïve IL1R-/- mice also failed to induce KC production to hapten challenge. These results indicate that IL1 signaling is required for optimal migration of hapten-presenting dendritic cells from the sensitized skin to the draining lymph nodes, for hapten-primed CD8 T cell priming, and for activation of these CD8 T cells by hapten-presenting endothelial cells in the challenge site to induce CXCL1/KC production and the CHS response.
ATP induced microvesicles stimulate macrophage activation (89.18)
Authors: Thomas Louis , Salter Russell 
Published: January 2023

Abstract
Adenosine triphosphate (ATP) has a unique role as a damage associated molecular pattern (DAMP) as it is normally kept within cells unless there is cellular/tissue injury. As a DAMP, ATP has several known mechanisms for immune cell activation. Our lab and others have reported that exogenous ATP administration to macrophages in cell culture induces rapid shedding of microvesicles (MV). In this study, we harvested MV from ATP treated macrophages and demonstrated that MV are capable of stimulating naive macrophages to release pro-inflammatory cytokines and induce co-stimulatory surface marker upregulation. MV- induced activation is dependent on a cAMP/p38 MAPK pathway and also results in NFkappaB translocation. Activation is partially dependent on TLR4, but is not due to the TLR4 agonist HMGB1, which is present in MV. TNFalpha is also present in MV but is not required to activate macrophages since MV from TNF-deficient cells are also stimulatory. Biochemical fractionation of MV was then used to show that the stimulatory component is phospholipid in nature, and we are currently working to identify the bioactive compound. These findings suggest a novel means by which macrophages at sites of cellular damage can become sterilely activated by ATP to further propagate cellular damage via shed microvesicles.
Induction of active immune suppression by co-immunization with DNA and cytokines plasmid in SPF chickens (89.26)
Authors: He Cheng , Ling Yong , Zhang Chang , Yang Junjing 
Published: January 2023

Abstract
Avian chlamydiosis has been widely prevalent in Chinese intensive farms, resulted in the significant economic loss as well as the huge potential threat to human being health. It is an urgent need to develop an appropriate vaccine against avian Chlamydopila psittaci. Th1-cell immunity plays a main role in the vaccination of avian Chlamydosis, but DNA vaccine based on the main outer membrane protein is unable to induce the incomplete protection in large animals. Hence, the co-vaccination with DNA and cytokine plasmids is considered to be a promising approach for improving efficacy of DNA vaccine. In this study, the SPF chickens were administered with DNA plus IL-2 plasmid, IL-12α plasmid, DNA-IL-12β plasmid and DNA-15 plasmid, respectively on day 0, day 14 and day 28. The birds inoculated with DNA were regarded as the positive control. All chickens were detected antibody levels,cytokine titers and challenged with C.psittaci to assess the protection. Results show that DNA vaccine plus four cytokine plasmids induce the significantly lower IgG antibody levels, the titers of IFN-γ and IL-12 as compared to DNA vaccine. Moreover, the spleens were observed to have heavy loads with chlamydia post challenge in four co-vaccination groups. The interesting findings indicate that DNA vaccine plus cytokine plasmids will induce the immune suppression. Keywords: omp A; Cytokine plasmid; immunosuppression; Chlamydophila psittaci
CD34+CD31+ immature myeloid cells (IMC) as an inhibitor cell on the production of antimicrobial peptide: III. Translational studies of glycyrrhizin in humanized NOD-SCID IL-2rγ-/- mice (NOD-SCID mice) (89.30)
Authors: Yoshida Shohei , Inatsu Akihito , Kobayashi Makiko , Herndon David , Suzuki Fujio 
Published: January 2023

Abstract
In the accompanying paper II, glycyrrhizin (GL) is shown to be inhibitory on the CD34+CD31+ IMC-mediated suppression of HBD-1 production by normal human epidermal keratinocytes (NHEK). In this study, the inhibitory effect of GL on CD34+CD31+ IMC-mediated suppression in HBD-1 production by NHEK was tested in human immune system mice (HIS mice) (X-irradiated NOD-SCID mice inoculated i.d. with NHEK and CD34+CD31+ IMC). Two days after the cell inoculation, inoculation site skin tissues were biopsied (8-mm diameter) from HIS mice treated with or without GL (Minophagen Pharmaceutical Co., Ltd., Tokyo, Japan) 6 and 24 hrs after the cell inoculation. Five biopsied tissue samples were homogenized together on ice in PBS. The concentrations of HBD-2 in the homogenates were measured by ELISA. In the results, 4.9 μg/ml of HBD-2 was detected in skin homogenates derived from NOD-SCID mice inoculated with NHEK, while HBD-2 was not detected in skin homogenates from HIS mice. When HIS mice were treated with 10 mg/kg of GL, HBD-2 was detected in IMC-inoculation site tissues of these mice. The amount of HBD-2 detected in this homogenate was corresponded to that detected in tissues of NOD-SCID mice inoculated with NHEK alone. These results indicate that GL improves the suppressed HBD-2 production in HIS mice. This study was supported by SHC #8610.
Plasma membrane-derived vesicles (PMVs) released from apoptotic Jurkat cells express upregulated surface Fas and induce apoptosis in recipient viable cells (89.45)
Authors: Jorfi Samireh , Ansa-Addo Ephraim , El dine Safa , Inal Jameel 
Published: January 2023

Abstract
Apoptosis, distinct from necrosis, is irreversible programmed cell death. Characterized by membrane blebbing, DNA fragmentation and cell shrinkage it plays an important role in infection. During viral infection, depending on the virus, apoptosis may either be enhanced or inhibited. Apoptosis can be induced by dexamethasone (DEX), cychloheximide (CHX) and H2O2 in vitro in various cell types. PMV release from apoptotic cells (isolated by differential centrifugation, expressing surface phosphatidylserine and of 0.2-1μm average diameter) could be inhibited by pretreatment with calpeptin. Such PMVs, which we distinguished from apoptotic bodies and other microvesicle populations, such as exosomes, have increased Fas expression on DEX and CHX- but not H2O2-mediated apoptotic Jurkat cells and an increased capacity to induce apoptosis in recipient healthy Jurkat cells, maximally at a 5:1 PMV-cell ratio. This effect, specific to PMVs from apoptotic cells, as opposed to those from viable cells, occurred after 30 min incubation of recipient, viable cells with PMVs and dose-dependently increased up to a maximum after 5h incubation. Inhibitors of caspase and anti-Fas blocking antibodies have been used to elucidate the importance of these factors in PMV transmission of this apoptotic signal. PMV-mediated apoptosis is currently being investigated for its role in in vitro studies of Coxsackie B3 infection in HeLa cells.
Galectin binding to and expression modulation by viruses in zebrafish (89.51)
Authors: Jackson Shawn , de las Heras Sanchez Ana , Ahmed Hafiz , Ammayappan Arun , Vakharia Vikram , Vasta Gerardo 
Published: January 2023

Abstract
The galectin family of β-galactoside-binding proteins is evolutionarily conserved. Galectins are known to interact with and affect the infectivity and/or pathogenicity of several viruses. We hypothesize that galectin interactions with viruses influence anti-viral immune responses. We investigate the interactions of zebrafish galectins with the glycosylated (G) protein of the rhabdovirus IHNV. We (a) characterized the carbohydrates present on IHNV proteins, and (b) identified direct interactions between zebrafish galectins (Drgal) and IHNV. We assessed the effects of immune challenge on (c) galectin and (d) cytokine expression in zebrafish. Several IHNV proteins exhibit complex glycosylation patterns. One or multiple IHNV proteins exhibit terminally linked mannose, sialic acid terminally linked to galactose, galactose-β(1-3)-GalNAc, sialic acid terminally linked to galactose or GalNAc, and galactose-β(1-4)-GlcNAc. The proto type Drgal1-L2, the chimera type Drgal3-L1 and the tandem-repeat type Drgal9-L2 all appear to bind to one or more IHNV proteins. All tested Drgal appear to bind to the IHNV G protein. ZFL cells endogenously express mRNA encoding Drgal1-L2, -L3, and -L4, Drgal3-L1 and -L2, and Drgal9-L1 mRNA. Drgal mRNA expression in ZFL cells was not induced by treatment with polyI:C. ZFL cells were found to endogenously express mRNA encoding Mx, TNFα, and IFNαβ. Treatment of these cells with polyI:C induced the expression of mRNA encoding IL-10 and IFNγ2.
Stimulation and priming of innate lymphocytes by oenothein B (89.59)
Authors: Ramstead Andrew , Schepetkin Igor , Quinn Mark , Jutila Mark 
Published: January 2023

Abstract
Oenothein B is an immunomodulatory tannin isolated from the Epilobium angustifolium plant. This compound is known to affect myeloid cells and these effects include enhanced chemotaxis, NF-κB activation, and production of proinflammatory cytokines, including TNFα and IL-6. Its effects on other innate immune cells have not been investigated. Previous studies have shown this compound to be beneficial in tumor models; therefore, it is of interest to determine what effect this compound may have on other innate immune cells known to be beneficial in anti-tumor responses, such as NK cells, NKT cells, and γδ T cells. Previous work from our lab has shown that certain tannins have the ability to directly stimulate γδ T cells, indicating oenothein B may also have immunomodulatory effects on this cell type. Our preliminary results show that oenothein B it is capable of stimulating γδ T cells and other lymphocytes. We also show that priming with oenothein B can enhance responses by these cells to secondary stimuli. This includes augmentation of IFNγ production, which may not be reliant upon oenothein B’s ability to stimulate both macrophages and γδ T cells. The molecular mechanisms, which facilitate these responses, are currently under investigation in both bovine and human cells. Funded by NIH Contract HHSN2662004000009/N01-AI40009, PO1 AT004986, and RR020185.
Beta-mannosylceramide represents a new class of NKT agonists that induces tumor immunity by a novel IFN-γ-independent mechanism. (89.7)
Authors: O'Konek Jessica , Illarionov Petr , Stewart Khursigara Deborah , Ambrosino Elena , Castillo II Bernard , Raju Ravinder , Khalili Maryam , Kim Hee-Yong , Howell Amy , Besra Gurdyal , Porcelli Steven , Berzofsky Jay , Terabe Masaki 
Published: January 2023

Abstract
Type I NKT cells are a unique lymphocyte population which recognize lipid antigens presented by the non-classical MHC-like molecule CD1d. Activation of type I NKT cells by α-galactosylceramide (α-GC) induces tumor immunity by IFN-γ dependent mechanisms. In this study, we characterized how modifications of the sugar head group of α-GC impacted tumor protection in an in vivo mouse model of lung metastasis. β-mannosylceramide (β-ManCer), but not α-mannosylceramide or α-fucosylceramide, induced strong type I NKT cell-dependent tumor immunity. Because this is the first report of a β-linked glycosylcermide capable of inducing significant type I NKT cell-mediated tumor protection, we further characterized the mechanism by which β-ManCer induced tumor elimination. Interestingly, unlike α-GC and its analogs which protected through IFN-γ dependent mechanisms, β-ManCer induced very little cytokine production both in vitro and in vivo and retained protection in the absence of IFN-γ. Rather, β-ManCer protected through a TNF-α and nitric oxide synthase-dependent mechanism novel for NKT cells. Simultaneous administration of sub-therapeutic doses of β-ManCer and α-GC resulted in synergistic reduction in tumor burden, supporting the conclusion that these glycolipids work through different mechanisms. The distinct mechanism of action of β-ManCer and its ability to synergize with α-GC suggest that β-ManCer is the first representative of a new class of type I NKT cell agonists.
Early myocardial infarction with concurrent complement response studied using a novel immunofluorescent approach (89.8)
Authors: Zhang Ming , Charlagorla Pradeepkumar , Green Danielle , Balroop Javi , Shaka-Idusuyi Oghomwen , Reza Mohammed , Patel Monaliben , Matta Sravan 
Published: January 2023

Abstract
Experimental myocardial infarction is typically determined using TTC. TTC, reduced to a red product in intact cells by endogenous enzymes/cofactors, remains colorless in damaged cells from which the enzymes/cofactors have been removed during reperfusion. Its use is limited to studies with prolonged reperfusion and sections cannot be used for other pathological events. Two recent studies showed that propidium iodide (PI), which enters damaged cells and intercalates with DNA to produce a red fluorescent complex, can be injected in vivo following ischemia/reperfusion to assess infarct size. We build on this in a murine model, using PI to study early infarct development and to evaluate the concurrent inflammatory response as indicated by complement C3 deposition. Occlusion of the coronary artery was followed by increasing periods of reperfusion and injection of PI to define infarction. Blue fluorescent microspheres were injected to define the area at risk (AAR). Heart slices were immediately imaged for infarction and AAR, and cryosections obtained from the slices imaged for C3 using FITC-labeled antibody. PI fluorescence revealed a significant infarction at 1 hour of reperfusion which was maintained through 24 hours. C3 deposition lagged infarction development, becoming significant at 3 hours of reperfusion. In summary, in vivo injection of PI allows study of the early stages of infarction and permits analysis, in the same tissue, of concurrently occurring inflammatory events.
Signalling pathways in the inflammatory process of a severe protrein malnutrition model (90.10)
Authors: Roux María , Calabrese Graciela 
Published: January 2023

Abstract
Previous work has shown that dietary antigen starts an intestinal inflammatory process, regulated by TNFα, inducing inflammatory genes transcription by NF-kB factor translocation into intestinal epitelial cells (IECs) nucleus . Therefore, we study IL-17 expression and its related transcriptional factors in our model of secondary immunodeficiency by malnutrition .Western Blot was used to analyze intracellular distribution ( microsomes ,nucleus and cytosol) obtained by ultracentrifugation of IECs from wellnourrished rats (C60) ,as well as from renourrished ones (weaning rats lost 25% of intially body weight and refed with a 20% casein diet during 21 days ( R21). Detection was performed with specific biotinylated antibodies to IL-17; ROR γt , NF-k B and STAT3 and avidin- biotin peroxidase system to detect the immunoreactive bands that were revealed with the DAB substrate. Results: IL-17 was found in the microsomal fraction accompagnied by a nuclear localization of ROR γt , NF-kB and STAT 3 only for R21 rats; while no differences in nuclear Foxp3 detection was observed between C60 compared with R21. We assume that the inflammatory process is associated with a nuclear localization of ROR γt, NF-kB and STAT3 in the renourrished animals withouth activation of Foxp3. Therefore, IL-17 expression contributes to the perpetuation of the inflammatory process, as downregulation of Foxp3 strongly correlates with upregulation of IL-17.
Dendritic cell-expressed CD103 is required for inhibition of mucosal Th2 cell responses (90.3)
Authors: Mullaly Sarah , Maltby Steven , Burrows Kyle , McNagny Kelly , Zaph Colby 
Published: January 2023

Abstract
Dysregulated mucosal CD4 Th2 cell responses are a feature of several inflammatory diseases including asthma, allergies and some forms of inflammatory bowel disease. The integrin CD103 is expressed primarily on a subset of dendritic cells (DCs) that promote regulatory T cell development and intestinal immune homeostasis. However, the role of CD103 in Th2-dependent immunity has not been tested. Using the helminth parasite Trichuris muris, we identify a critical role for CD103 in specifically limiting Th2 responses in vivo. We find that following infection with Trichuris, CD103-deficient (CD103-/-) mice develop an enhanced Th2 response that is associated with decreased levels of IL-10. While in vitro CD4 T cell differentiation was unaffected by CD103 deficiency, bone marrow-derived CD103-/- DCs induce heightened T cell proliferation and increased IL-13 production under Th2 promoting conditions. The inhibitory role of CD103 appears to be specific for Th2 responses, as CD103-/- mice are protected from disease development in a model of chemically-induced colitis. Taken together, these results suggest that CD103 expression on DCs is critically required to specifically regulate Th2 immune responses in vivo.
Shaping of the gut microbiota by B lymphocytes influences GATA4-dependent lipid absorption and body fat accumulation (90.8)
Authors: Shulzhenko Natalia , Morgun Andrey , Gavrilova Oksana , Battle Michele , Matzinger Polly 
Published: January 2023

Abstract
In order to understand the role for B lymphocytes in normal physiology of intestine, we compared global gene expression in the jejunum of wild-type (WT) and B-cell knockout (KO) mice. We found about 300 genes whose expression differed between WT and B-cell KO mice. Genes down-regulated in B- cell KO intestines, were highly enriched for lipid/energy metabolism genes. Consistent with this result, we found lower amounts of para-genital fat and serum leptin level in B-cell KO mice than in their B-cell-sufficient littermates. We revealed GATA4 as the most enriched transcription factor for the down regulated genes. Conditional KO of Gata4 in the intestinal epithelium showed a high correlation with BcKO gene signature. We also found a decrease in cholesterol absorption in the gut of B-cell KO mice (p&lt;0.005) previously observed in Gata4 intestinal KO. In addition, Gata4 KO mice perfectly recapitulated the systemic effects observed in B-cell KO mice presenting decreased levels of body fat and leptin (p&lt;0.01). Studying the mechanism, we found that mice that have B lymphocytes but do not produce soluble antibodies exhibit a very similar gene expression profile as the B-cell KO mice implying that this phenomenon is antibody-dependent. Under germ-free conditions, we found almost no difference between B-cell KO and control mice. Thus we conclude that B lymphocytes are involved in the control of GATA4-dependent lipid absorption and accumulation of body fat by shaping the gut microflora.