bluuebunny/crossref_metadata_2025_split huggingface dataset:
TCR Engagement Regulates Differential Responsiveness of Human Memory T Cells to Fas (CD95)-Mediated Apoptosis
Authors: Di Somma M. Maddalena , Somma Francesca , Montani Maria Saveria Gilardini , Mangiacasale Rosamaria , Cundari Enrico , Piccolella Enza 
Published: January 2023

Abstract
In this work, we have tried to establish whether human memory T cells may be protected from Fas (CD95)-induced apoptosis when correctly activated by Ag, and not protected when nonspecifically or incorrectly activated. In particular, we wanted to investigate the molecular mechanisms that regulate the fate of memory T cells following an antigenic challenge. To address this issue, we chose an experimental system that closely mimics physiological T cell activation such as human T cell lines and clones specific for viral peptides or alloantigens. We demonstrate that memory T cells acquire an activation-induced cell death (AICD)-resistant phenotype when TCRs are properly engaged by specific Ag bound to MHC molecules. Ag concentration and costimulation are critical parameters in regulating the protective effect. The analysis of the mechanisms involved in the block of CD95 signal transduction pathways revealed that the crucial events are the inhibition of CD95-associated IL-1β-converting enzyme (ICE)-like protease (FLICE) activation and poly(ADP)-ribose polymerase cleavage, and the mRNA expression of FLICE-like inhibitory protein. Furthermore, we have observed that TCR-mediated neosynthesis of FLICE-like inhibitory protein mRNA is suppressed either by protein tyrosine kinase inhibitors or cyclosporin A. In conclusion, the present analysis of the effects of TCR triggering on the regulation of AICD suggests that AICD could be inhibited in human memory T cells activated in vivo by a foreign Ag, but may become operative when the Ag has been cleared.
T1/ST2 Expression Is Enhanced on CD4+ T Cells from Schistosome Egg-Induced Granulomas: Analysis of Th Cell Cytokine Coexpression Ex Vivo
Authors: Löhning Max , Grogan Jane L. , Coyle Anthony J. , Yazdanbakhsh Maria , Meisel Christian , Gutierrez-Ramos Jose-Carlos , Radbruch Andreas , Kamradt Thomas 
Published: January 2023

AbstractTh cells are categorized into subsets based on the cytokine production of in vitro-differentiated Th populations. For in vivo-differentiated Th subsets, little is known about the heterogeneity of cytokine production in single cells. We recently described a molecule, T1/ST2, that is preferentially expressed on the surface of Th2 cells. Here we combined high-gradient magnetic cell separation with four-color single-cell cytometry to analyze simultaneously three intracellular cytokines and T1/ST2 surface expression on CD4+ cells from lungs containing granulomas induced by Schistosoma mansoni eggs. T1/ST2 was highly up-regulated on CD4+ T cells from hepatic granulomas and granulomatous lungs. T1/ST2+ cells from granulomatous lungs preferentially produced type 2 cytokines ex vivo. In the total CD4+ population, coexpression of type 1 and type 2 cytokines occurred frequently. However, such coproduction was drastically reduced in T1/ST2+ cells compared with T1/ST2− cells. Coexpression of type 1 and type 2 cytokines was also rare in cells simultaneously producing two cytokines of one type. These findings indicate that individual CD4+ T cells in vivo have different levels of commitment to a certain Th phenotype. Coexpression of two type 2 cytokines or production of one type 2 cytokine together with surface expression of T1/ST2 indicate advanced commitment to the Th2 phenotype.
Modulation of Monocytic Cell Activity and Virus Susceptibility During Differentiation into Macrophages
Authors: Basta Sameh , Knoetig Sonja M. , Spagnuolo-Weaver Martha , Allan Gordon , McCullough Kenneth C. 
Published: January 2023

Abstract
A major component of innate immune responses relies on monocytes and macrophages, virus infection of which will pose a particular problem for immunological defense. Consequently, the monocytic cell differentiation pathway was analyzed in terms of cellular modulations therein and their relation to monocytotropic virus infection. Differentiation was characterized by down-regulation of CD14, MHC Ags, the monocytic SWC1 marker, and p53; concomitant up-regulation of the SWC9 macrophage marker, a putative porcine CD80 (detected with anti-human CD80 Ab), and acid phosphatase secretion were also characteristic. Elevated phagocytic and endocytic activities as well as endosomal/lysosomal acidification were identified as being important to the macrophage. In contrast, monocytes possessed high accessory activity. This was multifactorial, concomitantly requiring 1) high MHC Ag expression; 2) enzyme activity of esterase, peroxidase, myeloperoxidase, and 5′ nucleotidase in preference to glucosidase, galactosidase, and glucuronidase; and 3) elevated capacity for spontaneous IL-1 production. Only with all parameters was efficient stimulation of Ag-specific lymphocytes possible. These results point to a continuous process during differentiation, involving inter-related characteristics linking the more accessory monocyte to the scavenger macrophage, both in vitro and in vivo. Of particular interest was how these characteristics related to monocytotropic virus infection, and how a particular virus could show a clear preference for the differentiating macrophages. Such results not only further our understanding of porcine immunology, but also provide evidence and a potential model for the determination and characterization of monocytotropic virus-host cell interactions.
Administration of an IL-12-Encoding DNA Plasmid Prevents the Development of Chronic Graft-Versus-Host Disease (GVHD)
Authors: Okubo Tadanobu , Hagiwara Eri , Ohno Shigeru , Tsuji Takashi , Ihata Atsushi , Ueda Atsuhisa , Shirai Akira , Aoki Ichiro , Okuda Kenji , Miyazaki Jun-ichi , Ishigatsubo Yoshiaki 
Published: January 2023

AbstractThe transfer of DBA/2 spleen cells into (C57BL/10 × DBA/2)F1 mice induces chronic graft-vs-host disease (GVHD), which is characterized by the production of Th2 cytokines, hypergammaglobulinemia, and immune complex-mediated glomerulonephritis like systemic lupus erythematosus. IL-12 strongly induces the production of Th1 cytokines and reduces Th2 activity in vivo. In this study, the effect of gene therapy on the development of murine chronic GVHD was examined using an IL-12-encoding plasmid (pCAGGSIL-12), with the expectation that it might regulate Th1/Th2 activity and have a beneficial impact on the clinical manifestations of disease. pCAGGSIL-12 or its p40 antagonist plasmid (pCAGGSp40) were injected i.m. every 3 wk in GVHD-induced (C57BL/10 × DBA/2)F1 mice. A total of 100 μg of pCAGGSIL-12 improved the Th1/Th2 balance in vivo, suppressed the production of IgG, and significantly reduced the development of glomerulonephritis. GVHD was exacerbated by injection of the pCAGGSp40 antagonist. Our results demonstrate that GVHD can be treated successfully by the administration of an IL-12-encoding plasmid, and that such therapy does not induce acute GVHD.
Similarities and Differences Between Human and Murine TNF Promoters in Their Response to Lipopolysaccharide
Authors: Kuprash Dmitry V. , Udalova Irina A. , Turetskaya Regina L. , Kwiatkowski Dominic , Rice Nancy R. , Nedospasov Sergei A. 
Published: January 2023

AbstractTranscription of the TNF gene is rapidly and transiently induced by LPS in cells of monocyte/macrophage lineage. Previous data suggested that multiple NF-κB/Rel binding sites play a role in the transcriptional response to LPS of the murine gene. However, the relevance of homologous sites in the human TNF gene remained a matter of controversy, partly because the high affinity NF-κB/Rel site located at −510 in the murine promoter is not conserved in humans. Here we used two sets of similarly designed human and mouse TNF promoter deletion constructs and overexpression of IκB in the murine macrophage cell line ANA-1 to show remarkable similarity in the pattern of the transcriptional response to LPS, further demonstrating the functional role of the distal promoter region located between −600 and −650. This region was characterized by mutagenesis of protein binding sites, including two relatively low affinity NF-κB/Rel sites, #2 and 2a. Mutation in each of the NF-κB sites resulted in 2- to 3-fold lower transcriptional activity in response to LPS. In contrast to LPS activation, the response to PMA was substantially lower in magnitude and required only the proximal promoter region. In summary, the functional topography of human and murine promoters when assayed in the same system has some marked similarities. Our observations support the notion that full LPS response of TNF gene requires both NF-κB and non-NF-κB nuclear proteins. Our data also suggest that the functional activity of a given κB site depends on the entire DNA sequence context in the promoter region.
Repression of IL-4-Induced Gene Expression by IFN-γ Requires Stat1 Activation
Authors: Venkataraman Chandrasekar , Leung Stewart , Salvekar Anupama , Mano Hiroyuki , Schindler Ulrike 
Published: January 2023

Abstract
IFN-γ antagonizes many physiological responses mediated by IL-4, including the inhibition of IL-4-induced IgE production. This event is largely mediated at the level of transcription. We observed that the IL-4 response element of the germline epsilon promoter is sufficient to confer IFN-γ-mediated repression onto a reporter construct. The inhibitory effects were observed in both lymphoid and nonlymphoid cell lines. Stat1, which is activated by IFN-γ, cannot recognize the Stat6-specific IL-4 response element in the ε promoter. Hence, competitive DNA binding does not seem to be the underlying mechanism for the inhibitory effect. This is supported by the observation that inhibition is not seen at early time points, but requires prolonged IFN-γ treatment. IFN-γ stimulation results in a loss of IL-4-induced Stat6 tyrosine phosphorylation, nuclear translocation, and DNA binding. Using the fibrosarcoma cell line U3A, which lacks Stat1, we demonstrated that the transcription activation function of Stat1 is required for the IFN-γ-mediated repression. Repression was restored by overexpression of Stat1α, but not Stat1β, in U3A cells. Treatment with IFN-γ, but not IL-4, specifically up-regulates the expression of SOCS-1 (silencer of cytokine signaling), a recently characterized inhibitor of cytokine signaling pathways, such as IL-6 and IFN-γ. Overexpression of SOCS-1 effectively blocks IL-4-induced Stat6 phosphorylation and transcription. This suggests that IFN-γ-mediated repression of IL-4-induced transcription is at least in part mediated by SOCS-1.
CD40 Ligand-CD40 Interaction Induces Chemokines in Cervical Carcinoma Cells in Synergism with IFN-γ
Authors: Altenburg Andreas , Baldus Stephan E. , Smola Hans , Pfister Herbert , Hess Sigrun 
Published: January 2023

Abstract
Cellular immunity plays a major role in controlling human papilloma virus infection and development of cervical carcinoma. Mononuclear cell infiltration possibly due to the action of chemokines becomes prominent in the tumor tissue. In fact, the macrophage chemoattractant protein-1, MCP-1, was detected in cervical squamous cell carcinoma in situ, whereas absent in cultured cells. From this, unknown environmental factors were postulated regulating chemokine expression in vivo. In this study, we show high CD40 expression on cervical carcinoma cells and CD40 ligand (CD40L) staining on attracted T cells in tumor tissue, suggesting a paracrine stimulation mechanism via CD40L-CD40 interactions. We therefore investigated chemokine synthesis in nonmalignant and malignant human papilloma virus-positive cell lines after CD40L exposure. Constitutive expression of MCP-1, MCP-3, RANTES, and IFN-γ-inducible protein-10 was almost undetectable in all cell lines tested. CD40L was able to induce MCP-1 production; however, despite much higher CD40 expression in malignant cells, MCP-1 induction was significantly lower compared with nontumorigenic cells. After sensitization with IFN-γ, another T cell-derived cytokine showing minimal effects on CD40 expression levels, CD40 ligation led to a more than 20-fold MCP-1 induction in carcinoma cell lines. An even stronger effect was observed for IFN-γ-inducible protein-10. Our study highlights the synergism of T cell-derived mediators such as CD40L and IFN-γ for chemokine responses in cervical carcinoma cells, helping to understand the chemokine expression patterns observed in vivo.
Overexpression of Bcl-2 in Transgenic Mice Decreases Apoptosis and Improves Survival in Sepsis
Authors: Hotchkiss Richard S. , Swanson Paul E. , Knudson C. Michael , Chang Katherine C. , Cobb J. Perren , Osborne Dale F. , Zollner Kimberly M. , Buchman Timothy G. , Korsmeyer Stanley J. , Karl Irene E. 
Published: January 2023

AbstractIn sepsis there is extensive apoptosis of lymphocytes, which may be beneficial by down-regulating the accompanying inflammation. Alternatively, apoptosis may be detrimental by impairing host defense. We studied whether Bcl-2, a potent antiapoptotic protein, could prevent lymphocyte apoptosis in a clinically relevant model of sepsis. Transgenic mice in which Bcl-2 was overexpressed in T cells had complete protection against sepsis-induced T lymphocyte apoptosis in thymus and spleen. Surprisingly, there was also a decrease in splenic B cell apoptosis in septic Bcl-2 overexpressors compared with septic HeJ and HeOuJ mice. There were marked increases in TNF-α, IL-1β, and IL-10 in thymic tissue in sepsis in the three species of mice, and the increase in TNF-α and IL-10 in HeOuJ mice was greater than that in Bcl-2 mice. Mitotracker, a mitochondrial membrane potential indicator, demonstrated a sepsis-induced loss of membrane potential in T cells in HeJ and HeOuJ mice but not in Bcl-2 mice. Importantly, Bcl-2 overexpressors also had improved survival in sepsis. To investigate the potential impact of loss of lymphocytes on survival in sepsis, Rag-1−/− mice, which are totally deficient in mature T and B cells, were also studied. Rag-1−/− mice had decreased survival compared with immunologically normal mice with sepsis. We conclude that overexpression of Bcl-2 provides protection against cell death in sepsis. Lymphocyte death may be detrimental in sepsis by compromising host defense.
Dose Dependence of CTL Precursor Frequency Induced by a DNA Vaccine and Correlation with Protective Immunity Against Influenza Virus Challenge
Authors: Fu Tong-Ming , Guan Liming , Friedman Arthur , Schofield Timothy L. , Ulmer Jeffrey B. , Liu Margaret A. , Donnelly John J. 
Published: January 2023

AbstractIntramuscular injection of BALB/c mice with a DNA plasmid encoding nucleoprotein (NP) from influenza virus A/PR/8/34 (H1N1) provides cross-strain protection against lethal challenge with influenza virus A/HK/68 (H3N2). CTL specific for the H-2Kd-restricted epitope NP147–155 are present in these mice and are thought to play a role in the protection. To assess the effectiveness of NP DNA immunization in comparison with influenza virus infection in the induction of CTL responses, we monitored the frequency of CTL precursors (CTLp) in mice following i.m. injection with NP DNA or intranasal infection with influenza virus and showed that the CTLp frequency in NP DNA-immunized mice can reach levels found in mice that had been infected with influenza virus. We also measured the CTLp frequency, anti-NP Ab titers, and T cell proliferative responses in mice that were injected with titrated dosages of NP DNA and documented a correlation of the CTLp frequency and the Ab titers, but not proliferative responses, with the injection dose. Furthermore, we observed a positive correlation between the frequency of NP147–155 epitope-specific CTLp and the extent of protective immunity against cross-strain influenza challenge induced by NP DNA injection. Collectively, these results and our early observations from adoptive transfer experiments of in vitro activated lymphocytes from NP DNA-immunized mice suggest a protective function of NP-specific CTLp in mice against cross-strain influenza virus challenge.
Extracellular Signal-Regulated Kinases Regulate Leukotriene C4 Generation, But Not Histamine Release or IL-4 Production from Human Basophils
Authors: Miura Katsushi , Schroeder John T. , Hubbard Walter C. , MacGlashan Donald W. 
Published: January 2023

AbstractHuman basophils secrete histamine and leukotriene C4 (LTC4) in response to various stimuli, such as Ag and the bacterial product, FMLP. IgE-mediated stimulation also results in IL-4 secretion. However, the mechanisms of these three classes of secretion are unknown in human basophils. The activation of extracellular signal-regulated kinases (ERKs; ERK-1 and ERK-2) during IgE- and FMLP-mediated stimulation of human basophils was examined. Following FMLP stimulation, histamine release preceded phosphorylation of ERKs, whereas phosphorylation of cytosolic phospholipase A2 (cPLA2), and arachidonic acid (AA) and LTC4 release followed phosphorylation of ERKs. The phosphorylation of ERKs was transient, decreasing to baseline levels after 15 min. PD98059 (MEK inhibitor) inhibited the phosphorylation of ERKs and cPLA2 without inhibition of several other tyrosine phosphorylation events, including phosphorylation of p38 MAPK. PD98059 also inhibited LTC4 generation (IC50 = ∼2 μM), but not histamine release. Stimulation with anti-IgE Ab resulted in the phosphorylation of ERKs, which was kinetically similar to both histamine and LTC4 release and decreased toward resting levels by 30 min. Similar to FMLP, PD98059 inhibited anti-IgE-mediated LTC4 release (IC50, ∼2 μM), with only a modest effect on histamine release and IL-4 production at higher concentrations. Taken together, these results suggest that ERKs might selectively regulate the pathway leading to LTC4 generation by phosphorylating cPLA2, but not histamine release or IL-4 production, in human basophils.
CD8 T Cells Are Essential in the Development of Respiratory Syncytial Virus-Induced Lung Eosinophilia and Airway Hyperresponsiveness
Authors: Schwarze Jürgen , Cieslewicz Grzegorz , Joetham Anthony , Ikemura Toshihide , Hamelmann Eckard , Gelfand Erwin W. 
Published: January 2023

AbstractViral respiratory infections can cause bronchial hyperresponsiveness and exacerbate asthma. In mice, respiratory syncytial virus (RSV) infection results in airway hyperresponsiveness (AHR) and eosinophil influx into the airways. The immune cell requirements for these responses to RSV infection are not well defined. To delineate the role of CD8 T cells in the development of RSV-induced AHR and lung eosinophilia, we tested the ability of mice depleted of CD8 T cells to develop these symptoms of RSV infection. BALB/c mice were depleted of CD8 T cells using anti-CD8 Ab treatment before intranasal administration of infectious RSV. Six days postinfection, airway responsiveness to inhaled methacholine was assessed by barometric body plethysmography, and numbers of lung eosinophils and levels of IFN-γ, IL-4, and IL-5 in bronchoalveolar lavage fluid were monitored. RSV infection resulted in airway eosinophilia and AHR in control mice, but not in CD8-depleted animals. Further, whereas RSV-infected mice secreted increased amounts of IL-5 into the airways as compared with noninfected controls, no IL-5 was detectable in both bronchoalveolar lavage fluid and culture supernatants from CD8-depleted animals. Treatment of CD8-depleted mice with IL-5 fully restored both lung eosinophilia and AHR. We conclude that CD8 T cells are essential for the influx of eosinophils into the lung and the development of AHR in response to RSV infection.
Lipopolysaccharide-Coated Erythrocytes Activate Human Neutrophils Via CD14 While Subsequent Binding Is Through CD11b/CD18
Authors: Troelstra Annet , de Graaf-Miltenburg Lia A. M. , van Bommel Toon , Verhoef Jan , Van Kessel Kok P. M. , Van Strijp Jos A. G. 
Published: January 2023

AbstractInteraction of LPS with monocytes and neutrophils is known to occur via CD14 and is strongly enhanced by LPS-binding protein (LBP). Integrins as well as CD14 play a role in the interaction of erythrocytes (E) coated with LPS or whole Gram-negative bacteria with phagocytes. We reasoned that the density of LPS on a particle is an important determinant in these interactions. Therefore, E were coated with different concentrations of LPS (ELPS). The binding of these ELPS to neutrophils was evaluated by flow cytometry. Simultaneously, we measured fMLP receptor expression to evaluate neutrophil activation. ELPS only bound to neutrophils in the presence of LBP. Blocking CD14 inhibited both activation and binding, whereas blocking complement (C) receptor 3 (CR3) inhibited binding but not activation. TNF activation restored ELPS binding in CD14-blocked cells but not in cells in which CR3 was blocked. Salmonella minnesota did bind to neutrophils independent of CR3 or CD14. The addition of LBP enhanced binding twofold, and this surplus was dependent upon CD14 but not on CR3. We conclude that ELPS interact with neutrophils via CD14, initially giving rise to cell activation; subsequently, binding is solely mediated by activated CR3.
Nuclear Factor-κB Mediates TNF-α Inhibitory Effect on α2(I) Collagen (COL1A2) Gene Transcription in Human Dermal Fibroblasts
Authors: Kouba David J. , Chung Kee-Yang , Nishiyama Takafumi , Vindevoghel Laurence , Kon Atsushi , Klement John F. , Uitto Jouni , Mauviel Alain 
Published: January 2023

Abstract
Among its plethora of activities as an inflammatory mediator, TNF-α has potent regulatory control on extracellular matrix production and degradation. Earlier studies have documented that TNF-α inhibits type I collagen gene (COL1A2) expression at the transcriptional level, but the characterization of the transcription factors involved has been elusive. In the present study, using transient cell transfection of human dermal fibroblasts with a battery of 5′ end deletion/chloramphenicol acetyltransferase (CAT) reporter gene constructs, we have characterized the TNF-α response element of the COL1A2 promoter. The TNF-α response element was attributed to a specific region that comprises noncanonical activator protein-1 (AP-1) (CGAGTCA) and NF-κB (AGAGTTTCCC) binding sites. TNF-α effect was eliminated by a 2-bp substitution mutation in the NF-κB1 binding half site of the NF-κB cis element. Electrophoretic mobility shift assays (EMSA) showed that recombinant human NF-κB heterodimers as well as NF-κB1 and RelA homodimers, but not AP-1, were capable of binding this element. Further, EMSA with human fibroblast nuclear extracts demonstrated enhanced binding of a single, specific complex within 5 min of TNF-α stimulation, which reached a plateau by 1 h and was not affected by preincubation of cells with cycloheximide. Gel supershift assays identified the complex as the NF-κB (p50/p65) heterodimer, whereas Abs to nuclear factor of activated T cells (NF-AT) and Jun family members failed to recognize the complex. These data suggest that in fibroblasts TNF-α activates and initiates the nuclear translocation of NF-κB that binds a divergent NF-κB element and plays a critical role in the observed inhibition of α2(I) collagen gene transcription.