bluuebunny/crossref_metadata_2025_split huggingface dataset:
Signaling Through the EBV/C3d Receptor (CR2, CD21) in Human B Lymphocytes: Activation of Phosphatidylinositol 3-Kinase via a CD19-Independent Pathway
Authors: Bouillie Sylvie , Barel Monique , Frade Raymond 
Published: December 2022

AbstractWe herein analyzed the regulation of phosphatidylinositol 3-kinase (PI 3-kinase) activity by CR2 activated on B lymphocyte cell surface. We demonstrated that CR2 activation triggered in vivo PI 3-kinase activity and interaction of PI 3-kinase p85 subunit with a tyrosine-phosphorylated p95 component. The specificity of PI 3-kinase activity was controlled using wortmannin and LY294002. CR2 activation did not trigger tyrosine phosphorylation of PI 3-kinase p85 subunit, but induced direct interaction of tyrosine phosphorylated p95 with the Src homology 2 domain of p85 subunit, as shown using glutathione-S-transferase fusion proteins. Despite identical molecular masses, immunoblotting analysis demonstrated that tyrosine-phosphorylated p95 that interacted in vivo and in vitro with p85 was neither CD19, the 95-kDa proto-oncogene vav, nor Gab1 (a 95-kDa adaptor molecule). Furthermore, p95 tyrosine phosphoprotein also expressed in K562A cells (CR2+ CD19− cells) interacted with Src homology 2 domain of PI 3-kinase p85 subunit after CR2 activation. Activated CR2 did not interact directly with p85 subunit or tyrosine-phosphorylated p95. This suggests the presence of an intermediate molecule between activated CR2 and tyrosine-phosphorylated p95, which may be 3BP2. In addition, in contrast to CD19 activation, CR2 activation did not trigger interaction of CD19 or Vav with PI 3-kinase p85 subunit or coprecipitation of PI 3-kinase activity with CD19. Together, these data clearly demonstrated that CR2 activation triggered in vivo PI 3-kinase activation through a pathway distinct from that triggered through CD19 activation.
Immunosuppressive Activities of Recombinant Glycosylation -Inhibiting Factor Mutants
Authors: Tomura Takafumi , Watarai Hiroshi , Honma Nakayuki , Sato Masahiro , Iwamatsu Akihiro , Kato Yoichi , Kuroki Ryota , Nakano Tatsumi , Mikayama Toshifumi , Ishizaka Kimishige 
Published: December 2022

AbstractWe have shown previously that glycosylation-inhibiting factor (GIF) in culture supernatants of suppressor T cell (Ts) hybridomas had bioactivity, while the same cells contained a substantial quantity of inactive GIF in cytosol. Mass-spectrometric analysis of GIF in the culture supernatant and cytosol of a Ts hybridoma provided direct evidence that GIF protein was posttranslationally modified in the Ts cells, and that the GIF bioactivity is associated with the posttranslationally modified species. Assuming that conformational changes induced by the posttranslational modifications are responsible for generation of bioactivity, we constructed cysteine mutants of human rGIF (rhGIF) in which cysteine at position 57, 60, or 81 was replaced with Ala, and the mutants were expressed in Escherichia coli. Replacement of Cys57 or Cys60 with Ala resulted in generation of bioactivity, while replacement of Cys81 with Ala failed to do so. It was also found that replacement of Cys57 with Ala and carboxymethylation of a sulfhydryl group in Cys60 synergistically increased the GIF bioactivity of the GIF derivatives. A mutated GIF protein, in which Cys57 and Asn106 in the rhGIF were replaced with Ala and Ser, respectively, had immunosuppressive effects on the IgE and IgG1 Ab responses of BDF1 mice to DNP-OVA, while wild-type rhGIF did not. Evidence was obtained that the mutated GIF suppressed Ag priming of Th cells for the Ab responses and proliferative response.
IL-4 Enhances IL-10 Gene Expression in Murine Th2 Cells in the Absence of TCR Engagement
Authors: Schmidt-Weber C. B. , Alexander S. I. , Henault L. E. , James L. , Lichtman A. H. 
Published: December 2022

AbstractBoth IL-4 and IL-10 are regulatory cytokines produced by Th2 cells that can down-regulate cell-mediated immune responses. The studies reported here examine the influence of various cytokines in the regulation of T cell IL-10 production. The results indicate that IL-10 gene expression by TCR transgenic Th2 cells is significantly up-regulated by IL-4 in the absence of TCR signals. IL-4 enhances both IL-10 mRNA levels and secreted protein, and this effect is not related to enhanced mRNA stability. TCR-mediated IL-10 gene expression is inhibited by cyclosporin A, but IL-4-mediated IL-10 expression is not. IL-4 also enhances IL-13 mRNA levels, to a lesser extent than IL-10, but does not significantly effect the expression of other cytokine mRNAs. Furthermore, IL-4 does not significantly enhance IL-10 expression in Th1 cells. IL-2 also enhances effector cytokine production in the absence of TCR signals, but in a subset nonspecific manner, increasing both Th2 IL-4 mRNA and Th1 IFN-γ mRNA. These data suggest that Th2 IL-4 production may contribute to the down-regulation of immune responses by directly enhancing Th2 IL-10 production. In addition, the data clearly demonstrate that exogenous cytokines can significantly influence effector cytokine production by effector T cells without the requirement for TCR signals.
IL-2 Induces STAT4 Activation in Primary NK Cells and NK Cell Lines, But Not in T Cells
Authors: Wang Kathy S. , Ritz Jerome , Frank David A. 
Published: December 2022

AbstractIL-2 exerts potent but distinct functional effects on two critical cell populations of the immune system, T cells and NK cells. Whereas IL-2 leads to proliferation in both cell types, it enhances cytotoxicity primarily in NK cells. In both T cells and NK cells, IL-2 induces the activation of STAT1, STAT3, and STAT5. Given this similarity in intracellular signaling, the mechanism underlying the distinct response to IL-2 in T cells and NK cells is not clear. In this study, we show that in primary NK cells and NK cell lines, in addition to the activation of STAT1 and STAT5, IL-2 induces tyrosine phosphorylation of STAT4, a STAT previously reported to be activated only in response to IL-12 and IFN-α. This activation of STAT4 in response to IL-2 is not due to the autocrine production of IL-12 or IFN-α. STAT4 activated in response to IL-2 is able to bind to a STAT-binding DNA sequence, suggesting that in NK cells IL-2 is capable of activating target genes through phosphorylation of STAT4. IL-2 induces the activation of Jak2 uniquely in NK cells, which may underlie the ability of IL-2 to activate STAT4 only in these cells. Although the activation of STAT4 in response to IL-2 occurs in primary resting and activated NK cells, it does not occur in primary resting T cells or mitogen-activated T cells. The unique activation of the STAT4-signaling pathway in NK cells may underlie the distinct functional effect of IL-2 on this cell population.
A Selective Defect of IFN-γ- But Not of IFN-α-Induced JAK/STAT Pathway in a Subset of U937 Clones Prevents the Antiretroviral Effect of IFN-γ Against HIV-1
Authors: Bovolenta Chiara , Lorini Alessandro L. , Mantelli Barbara , Camorali Laura , Novelli Francesco , Biswas Priscilla , Poli Guido 
Published: December 2022

AbstractIFN-γ induces transcription of several IFN-stimulated genes (ISGs). Recently, the IFN-γ-dependent Janus kinase (JAK)/STAT pathway has been shown to mediate the activation of some ISGs, by the sequential phosphorylation of two JAK kinases (JAK1 and JAK2) and of STAT1. Given that the JAK/STAT is the major, but not the only pathway linked to the IFN-γR, aim of our work was to investigate the signal-transduction pathway(s) by which IFN-γ exerts its effects on acute replication of HIV in monocytic cells. To this end, we utilized clones previously derived from the U937 promonocytic cell line, differing for their efficient (plus clones) or inefficient (minus clones) abilities of supporting HIV replication. Unlike IFN-α, IFN-γ did not inhibit HIV replication in plus clones, whereas virus production in minus cells was efficiently inhibited by both types of IFN. Plus clones generated a JAK/STAT signal-transduction pathway in response to IFN-α, but not IFN-γ. In contrast, minus clones responded to either cytokines. The functional defect of plus clones in response to IFN-γ was correlated to a selective defect of IFN-γR2, but not IFN-γR1, membrane expression. Surprisingly enough, IFN-γ stimulation of plus clones induced IFN-stimulated gene factor 3 (ISGF3γ). These results strongly support the hypothesis that the JAK/STAT pathway is responsible for the antiretroviral effect of IFN-γ, and further provide evidence for a potential second pathway triggered by IFN-γ in the absence of IFN-γR2 chain cell surface expression and involving ISGF3γ.
Activation of Human CD8+ αβ TCR+ Cells byMycobacterium tuberculosisVia an Alternate Class I MHC Antigen-Processing Pathway
Authors: Canaday David H. , Ziebold Christine , Noss Erika H. , Chervenak Keith A. , Harding Clifford V. , Boom W. Henry 
Published: December 2022

AbstractHuman immune responses to M. tuberculosis are characterized by activation of multiple T cell subsets including CD4+, CD8+, and γδ T cells, and the role of CD8+ αβ TCR+ T cells in this response is poorly understood. Stimulation of T cells from healthy tuberculin skin test-positive persons with live M. tuberculosis-H37Ra or soluble M. tuberculosis Ags readily up-regulated IL-2Rα (CD25) expression on CD8+ T cells. Purified resting and activated CD8+ T cells produced IFN-γ and proliferated to both M. tuberculosis bacilli and soluble mycobacterial Ags with monocytes as APC. Precursor frequency of mycobacterial Ag-specific CD8+ T cells by IFN-γ enzyme-linked immunospot was 5–10-fold lower than the precursor frequency of CD4+ T cells, and IFN-γ secretion by CD8+ T cells was 50–100-fold lower. CD8+ T cells secreted ∼10-fold less IFN-γ per cell than CD4+ T cells in response to mycobacterial Ags. CD8+ T cell responses to M. tuberculosis bacilli were blocked by anti-MHC class I antibody and required Ag processing. Processing of M. tuberculosis bacilli by monocytes for presentation to MHC class I-restricted CD8+ T cells was insensitive to brefeldin A treatment, which blocks the conventional MHC class I Ag-processing pathway. These results represent the first demonstration that human cells can process pathogen Ags via an alternate Ag-processing pathway for MHC class I and suggest a mechanism for participation of IFN-γ-secreting CD8+ T cells in the human immune responses to M. tuberculosis.
Evaluation of the Role of Mitogen-Activated Protein Kinases in the Expression of Inducible Nitric Oxide Synthase by IFN-γ and TNF-α in Mouse Macrophages
Authors: Chan Edward D. , Winston Brent W. , Uh Soo-Taek , Wynes Murry W. , Rose David M. , Riches David W. H. 
Published: December 2022

AbstractThe expression of inducible nitric oxide synthase (iNOS) by macrophages is stimulated by coexposure to IFN-γ and a number of stimuli, including TNF-α. Recent work has shown that TNF-α activates members of the mitogen-activated protein kinase family that subsequently trans-activate transcription factors implicated in the regulation of iNOS expression. The objective of this study was to systematically evaluate the role of: 1) p42mapk/erk2, 2) p46 c-Jun NH2-terminal kinase/stress-activated protein kinase (p46 JNK/SAPK), and 3) p38mapk in the induction of iNOS expression during costimulation of mouse macrophages with IFN-γ and TNF-α. All three kinases were activated during costimulation with IFN-γ and TNF-α. However, specific antagonism of the p42mapk/erk2 and p38mapk with PD98059 and SKF86002, respectively, had no effect on the induction of iNOS expression. In contrast, blockade of all three kinases with N-acetylcysteine completely blocked the induction of iNOS expression. In addition, specific antagonism of the JNK/SAPK upstream kinases MEKK (mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase) and MKK4 (mitogen-activated protein kinase kinase 4) with dominant inhibitory mutants blocked transcriptional activation of the iNOS promoter in response to costimulation with IFN-γ and TNF-α. Collectively, these findings support the involvement of p46 JNK/SAPK and its upstream kinases in regulating the induction of iNOS following ligation of the TNF-α receptor CD120a (p55) in the presence of IFN-γ.
Regulation of T Cell Activation In Vitro and In Vivo by Targeting the OX40-OX40 Ligand Interaction: Amelioration of Ongoing Inflammatory Bowel Disease with an OX40-IgG Fusion Protein, But Not with an OX40 Ligand-IgG Fusion Protein
Authors: Higgins Lisa M. , McDonald Stuart A. C. , Whittle Nigel , Crockett Nigel , Shields John G. , MacDonald Thomas T. 
Published: December 2022

AbstractOX40 is a member of the TNFR superfamily, and is found predominantly on activated CD4-positive T cells. In vitro an OX40-IgG fusion protein inhibits mitogen- and Ag-driven proliferation and cytokine release by splenocytes and lymph node T cells. In contrast, an OX40 ligand-IgG fusion protein enhanced proliferative responses. In normal mice, OX40-positive cells are observed only in lymphoid tissues, including Peyer’s patches of the gut. In mice with hapten-induced colitis or IL-2 knockout mice with spontaneous colitis, OX40-positive cells are found infiltrating the lamina propria. Administration of the OX40-IgG fusion protein to mice with ongoing colitis (but not the OX40 ligand-IgG) ameliorated disease in both mouse models of inflammatory bowel disease. This was evidenced by a reduction in tissue myeloperoxidase; reduced transcripts for TNF-α, IL-1, IL-12, and IFN-γ; and a reduction in the T cell infiltrate. Targeting OX40 therefore shows considerable promise as a new strategy to inhibit ongoing T cell reactions in the gut.
IL-17 Is Produced by Nickel-Specific T Lymphocytes and Regulates ICAM-1 Expression and Chemokine Production in Human Keratinocytes: Synergistic or Antagonist Effects with IFN-γ and TNF-α
Authors: Albanesi Cristina , Cavani Andrea , Girolomoni Giampiero 
Published: December 2022

AbstractIL-17 is a novel T cell-derived cytokine that can regulate the functions of a variety of cell types. In this study, we investigated whether hapten-specific T cells isolated from patients with allergic contact dermatitis (ACD) to nickel produce IL-17 and the effects of IL-17 alone or in combination with IFN-γ or TNF-α on the immune activation of keratinocytes. Skin affected with ACD to nickel and skin-derived, nickel-specific CD4+ T cell lines expressed IFN-γ, TNF-α, and IL-17 mRNAs. Four of seven nickel-specific CD4+ T cell clones positive for the skin-homing receptor, cutaneous lymphocyte-associated Ag, were shown to corelease IL-17, IFN-γ, and TNF-α. In contrast, two nickel-specific CD8+ T cell clones failed to synthesize IL-17. Normal human keratinocytes were found to express constitutively the IL-17 receptor gene. IL-17 specifically and dose-dependently augmented IFN-γ-induced ICAM-1 expression on keratinocytes at both the mRNA and the protein level, whereas HLA-DR, MHC class I, and CD40 levels were not modulated by IL-17. On the other hand, IL-17 alone did not affect ICAM-1 or enhance TNF-α-induced ICAM-1. In addition, IL-17, both directly and in synergism with IFN-γ and/or TNF-α, stimulated synthesis and release of IL-8 by keratinocytes. In contrast, IFN-γ- and TNF-α-induced production of RANTES was markedly inhibited by IL-17, and the synthesis of macrophage chemotactic protein 1 was not changed. Taken together, the results suggest that IL-17 is an important player of T cell-mediated skin immune responses, with synergistic or antagonist effects on IFN-γ- and TNF-α-stimulated keratinocyte activation.
Cutting Edge: Human Myeloid Cells Express an Activating ILT Receptor (ILT1) That Associates with Fc Receptor γ-Chain
Authors: Nakajima Hideo , Samaridis Jacqueline , Angman Lena , Colonna Marco 
Published: December 2022

AbstractIg-like transcripts (ILTs) encode cell surface receptors expressed on myeloid and lymphoid cells that are structurally and functionally related to killer cell inhibitory receptors. One ILT, designated ILT1, contains a short cytoplasmic domain that lacks sequence motifs implicated in signal transduction. Its function is unknown. Similar short cytoplasmic domains have been observed in activating NK cell receptors and FcαR, which transduce stimulatory signals via associated DAP12 and FcεRIγ proteins, respectively. Here we show that ILT1 receptor is selectively expressed on myeloid cells, functions as an activating receptor, and associates with FcεRIγ rather than DAP12.
IFN-γ Increases the Severity and Accelerates the Onset of Experimental Autoimmune Uveitis in Transgenic Rats
Authors: Egwuagu Charles E. , Sztein Jorge , Mahdi Rashid M. , Li Wenmei , Chao-Chan Chi , Smith Janine A. , Charukamnoetkanok Puwat , Chepelinsky Ana B. 
Published: December 2022

AbstractExperimental autoimmune uveitis (EAU) is a predominantly Th1-mediated intraocular inflammatory disease that serves as a model for studying the immunopathogenic mechanisms of uveitis and organ-specific autoimmune diseases. Despite the well-documented role of IFN-γ in the activation of inflammatory cells that mediate autoimmune pathology, recent studies in IFN-γ-deficient mice paradoxically show that IFN-γ confers protection from EAU. Because of the implications of these findings for therapeutic use of IFN-γ, we sought to reexamine these results in the rat, another species that shares essential immunopathologic features with human uveitis and is the commonly used animal model of uveitis. We generated transgenic rats (TR) with targeted expression of IFN-γ in the eye and examined whether constitutive ocular expression of IFN-γ would influence the course of EAU. We show here that the onset of rat EAU is markedly accelerated and is severely exacerbated by IFN-γ. In both wild-type and TR rats, we found that the disease onset is preceded by induction of ICAM-1 gene expression and is characterized by selective recruitment of T cells expressing a restricted TCR repertoire in the retina. In addition, these events occur 2 days earlier in TR rats. Thus, in contrast to the protective effects of IFN-γ in mouse EAU, our data clearly show that intraocular secretion of IFN-γ does not confer protection against EAU in the rat and suggest that IFN-γ may activate distinct immunomodulatory pathways in mice and rats during uveitis.
High Frequency of Specific CD8+ T Cells in the Tumor and Blood Is Associated with Efficient Local IL-12 Gene Therapy of Cancer
Authors: Fernandez Nadine C. , Levraud Jean-Pierre , Haddada Hédi , Perricaudet Michel , Kourilsky Philippe 
Published: December 2022

AbstractCancer immunotherapy often aims at the reactivation and expansion of tumor-specific CTL. In an attempt to correlate in situ and/or systemic tumor-specific T cell expansion with tumor regression, we investigated the effects of adenovirus-mediated IL-12 or IFN-γ gene transfer into established P815 murine tumors. While IFN-γ was no more potent than the vector alone, IL-12 gene transfer promoted tumor eradication. Despite this antitumor effect, no significant cytolytic activity was detectable using classical cytotoxicity assays from in vitro restimulated splenocytes. Since intratumor gene delivery may induce a localized expansion of CTL, the presence of P815-specific CD8+ T cells in situ was assessed. Using the Immunoscope approach, we found a dramatic increase in clonotypic T cells at the tumor site following IL-12, but not IFN-γ gene delivery. Antitumor CD8+ T cell frequencies were then re-evaluated using this molecular detection technique, which revealed a comparable expansion of specific T cells in the peripheral organs, most strikingly in the blood. These data show that local IL-12 gene transfer, in contrast to IFN-γ, mediates a potent antitumor effect that correlates to clonal tumor-specific T cell expansions in situ and in the periphery.
TNF Recruits TRADD to the Plasma Membrane But Not thetrans-Golgi Network, the Principal Subcellular Location of TNF-R1
Authors: Jones Sally J. , Ledgerwood Elizabeth C. , Prins Johannes B. , Galbraith Jenny , Johnson David R. , Pober Jordan S. , Bradley John R. 
Published: December 2022

AbstractThe subcellular localization of TNF-R1 to the Golgi apparatus, initially observed in endothelial cells, has been confirmed using transfection of bovine aortic endothelial cells with a human TNF-R1 expression plasmid. The subcellular interactions of TNF-R1 and the TRADD (TNFR-associated death domain protein) adaptor protein have been analyzed in the human monocyte cell line U937 and the human endothelial cell line ECV304 by confocal immunofluorescence microscopy and by Western blot analysis of fractionated cell extracts. In untreated cells, in which TNF-R1 is found on the cell surface but principally localizes to the trans-Golgi network, TRADD is concentrated in the cis- or medial-Golgi region, but separates from the Golgi during cell fractionation. Coimmunoprecipitation studies have shown that TRADD binds to TNF-R1 within 1 min of TNF treatment in a cell fraction-containing plasma membrane. This association is followed by a gradual dissociation, which is prevented if receptor-mediated endocytosis is inhibited by hypertonic medium. In contrast, no association is detected between TRADD and TNF-R1 in the Golgi in response to exogenous TNF at any time examined. These results suggest that although TNF-R1 is predominantly a Golgi-associated protein and TRADD also localizes to the Golgi region, exogenous TNF causes TRADD to bind to TNF-R1 only at the plasma membrane.
IgE Hyperproduction Through Enhanced Tyrosine Phosphorylation of Janus Kinase 3 in NC/Nga Mice, a Model for Human Atopic Dermatitis
Authors: Matsumoto Masahiro , Ra Chisei , Kawamoto Keiko , Sato Hiroaki , Itakura Atsuko , Sawada Junko , Ushio Hiroko , Suto Hajime , Mitsuishi Kouichi , Hikasa Yoshiaki , Matsuda Hiroshi 
Published: December 2022

AbstractIgE hyperproduction frequently observed in patients with atopic dermatitis (AD) may greatly contribute to the pathogenesis of AD, but its mechanisms are still unclear. NC/Nga mice raised in nonsterile circumstances spontaneously suffered from AD-like skin lesions with elevation of plasma IgE levels. We investigated mechanisms of the IgE hyperproduction in NC/Nga mice. Splenic T cells from SPF NC/Nga mice had a level of CD40 ligand (CD40L) expression comparable to that of BALB/c mice. Although there was no difference in the expression of CD40 on B cells between NC/Nga and BALB/c mice, B cells of NC/Nga mice produced much more IgE in the presence of soluble CD40L and IL-4. The stimulation with CD40L and/or IL-4 resulted in tyrosine phosphorylation of Janus kinase 3 (JAK3) in B cells, which was more strongly inducible in NC/Nga mice than in BALB/c mice. In B cells isolated from PBMC of AD patients with high serum IgE levels, JAK3 was constitutively phosphorylated at the tyrosine residue, and its phosphorylation was enhanced by the treatment with CD40L and/or IL-4 as was that in splenic B cells of NC/Nga mice with dermatitis and high IgE levels. Thus, it is suggested that constitutive and enhanced JAK3 phosphorylation in B cells highly sensitive to CD40L and IL-4 may be attributable to IgE hyperproduction in NC/Nga mice and patients with AD.