bluuebunny/crossref_metadata_2025_split huggingface dataset:
Activation of cytokine production and adhesion molecule expression on THP-1 myelomonocytic cells by macrophage colony-stimulating factor in combination with interferon-γ
Authors: Kimball Edward S , Kovacs Elizabeth , Clark M Christina , Schneider Craig R 
Published: January 2018

Abstract
THP-1 myelomonocytic leukemia cells cultured with either macrophage colony-stimulating factor (M-CSF) or interferon-γ (IFN-γ) alone produce, at best, only low levels of interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α). However, combinations of the two factors resulted in at least 3- to 20-fold greater amounts of IL-1β and TNF-α than would have been predicted by additive mechanisms. This enhanced cytokine production was observed when M-CSF and IFN-γ were added simultaneously or when M-CSF was added 24 h after addition of IFN-γ to the cells. Similar results were obtained with fresh human peripheral blood cells treated with IFN-γ + M-CSF. Cycloheximide treatment of the cultures containing M-CSF and IFN-γ inhibited the production of IL-1β and TNF-α. Northern blotting studies revealed no effect of IFN-γ alone on IL-1β or TNF-α mRNA production. IL-1β and TNF-α mRNA expression was observed at 2 and 6 h after treatment with M-CSF or IFN-γ + M-CSF. Higher TNF-α mRNA expression was observed at 2 and 6 h after treatment with IFN-γ + M-CSF, and higher IL-1β mRNA expression was observed at 2 h after treatment with IFN-γ + M-CSF compared with mRNA levels observed for cells cultured only with M-CSF. These results suggest that the augmented cytokine production resulting from treatments with combinations of M-CSF and IFN-γ occurs due to increased cytokine mRNA and increased cytokine protein synthesis. In addition to up-regulating cytokines, combinations of IFN-γ and M-CSF resulted in augmented cell surface expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1. This was accompanied by morphological and functional changes that included plastic adherence, extensive homotypic aggregation, and a macrophage-like appearance. These phenotypic changes and enhancements in cytokine expression and cell surface molecule expression may be related to activation of monocytic cells to become cytotoxic effectors by M-CSF and IFN-γ combinations. In vitro cytotoxicity against A-375 melanoma cells was greatest for cultures that contained M-CSF and IFN-γ in combination.
Some aspects of IL-8 pathophysiology III: chemokine interaction with endothelial cells
Authors: Rot Antal , Hub Elin , Middleton James , Pons Françoise , Rabeck Christa , Thierer Kamillo , Wintle Jonathan , Wolff Barbara , Zsak Marion , Dukor Peter 
Published: January 2018

Abstract
Chemolrines have been convincingly implicated in driving leukocyte emigration in different inflammatory reactions. However, the cellular and molecular mechanisms of chemokine involvement in leukocyte emigration are not yet clear. We and others suggested that leukocyte adhesion to the endothelium and transmigration are induced by chemokines immobilised on the endothelial cell surface. This would require the presence of specific chemokine binding sites in this microanatomical location. Using an in situ binding assay we demonstrated the presence of binding sites for interleukin-8 (IL-8) and RANTES, but not monocyte inflammatory protein-1α on the endothelium of postcapillary venules and small veins in human skin. In contrast, venules and veins in various anatomical locations showed dramatically differing IL-8 binding patterns. The subcellular distribution of IL-8 in the venular endothelial cells following its in vivo and ex vivo injections was studied by use of electron microscopy. Our results suggest that IL-8 was internalized by the endothelial cells, transported transcellularly via plasmalemmal vesicles, and released onto the luminal surface where it appeared located preferentially on tips of membrane protrusions. We were unable to study the endothelial IL-8 binding or transport in vitro because all the in vitro propagated endothelial cell lines and primary endothelial cells tested lacked IL-8 binding sites. This includes human umbilical vein endothelial cells (HUVECs), which also did not bind IL-8 in situ. However, HUVECs provided a satisfactory in vitro system to study the secretion of IL-8 by the endothelial cells. Two possible alternative pathways were described: secretion directly from the Golgi apparatus or following storage in Weibel-Palade bodies.
Role of eosinophil-chemotactic C-C chemokines in cutaneous inflammation
Authors: Schröder Jens-M , Noso Norio , Sticherling Michael , Christophers Enno 
Published: January 2018

Abstract
In the dermal sites of atopic skin, eosinophil (Eo) granule protein or more rarely intact Eos represent a characteristic histological feature. We addressed the question of whether lesional scales of patients with various eosinophilic skin disorders contain Eo attractant and tried to characterize it biochemically. In scales of a patient with drug reaction, heparin-binding Eo attractants could be identified. High-performance liquid chromatographic analyses together with specific ELISA and Western blot analyses revealed identity with RANTES. No other heparin-binding Eo chemotaxin could be identified. HPLC analysis of pooled lesional scale extracts of patients with atopic dermatitis showed fractions containing only weak heparin-binding Eo-chemotactic activity, which, however, showed RANTES immunoreactivity. In experiments to elucidate the putative cellular origin of Eo-attracting chemokines in human skin we investigated supernatants of atopic skin–derived T lymphocytes as well as supernatants of stimulated dermal fibroblasts for Eo-chemotactic factors. Unexpectedly, we did not find any heparin-bound Eo attractants in supernatants of stimulated cultured atopic skin–derived T lymphocyte clones, whereas fibroblasts produced RANTES as well as granulocyte-macrophage colony-stimulating factor. Therefore, fibroblasts are a likely source of eosinophil attractant cells, which could contribute to the Eo infiltrate. Selectivity of the infiltrate might come from selective induction of RANTES and/or induction of other as yet unidentified Eo-specific chemokines.
Effects of dexamethasone* on bovine circulating T lymphocyte populations
Authors: Burton Jeanne L , Kehrli Marcus E 
Published: January 2018

Abstract
In cattle, γδ T cells represent a higher proportion of circulating T cells than in humans. Bovine γδ T cells can be recognized by expression of γδ T cell receptor (γδ TCR) determinants or by a 215/230-kDa surface antigen (WC1). WC1 is expressed on 90% or more of circulating bovine γδ T cells. The effects of dexamethasone on this and other subsets (CD3, CD2, CD4, CD8) of peripheral blood T lymphocytes were determined by flow cytometric analysis. Twelve 15-month old bulls were injected with dexamethasone (0.04 mg/kg/day) for 3 consecutive days and four bulls were untreated controls. Blood samples were collected daily for 3 days before dexamethasone injections and for an additional 7 days starting on the third injection day. Data were recorded as percent positive cells and as mean fluorescent intensity (MFI) of positive cells. Initially, CD3+ cells represented 65–73% of all peripheral blood mononuclear cells (PBMC). Dexamethasone reduced CD3+ cells to 30%, and these recovered to 50% positive cells by 9 days after the last dexamethasone injection. Loss of CD3+ cells was not due to reductions in αβ T cells because dexamethasone did not influence the percent CD2+, CD4+, or CD8+ cells. However, percent WC1+ cells rapidly declined from a baseline of 26.4% of PBMC to &lt; 6% by the final injection. During injections, the MFI of WC1 increased. The MFI of WC1 returned to control values 7 days after the last injection of dexamethasone, but the percent γδ T cells recovered to only 14% WC1 + PBMC by the final day of the study. During its maximum effects on WC1, dexamethasone also caused a profound decrease of L-selectin MFI on remaining PBMC (mostly αβ T cells and monocyte/macrophages). In a second trial, two-color analyses determined that dexamethasone did not increase apoptosis in WC1 + cells and did not reduce L-selectin MFI on either CD2+ or WC1 + cells. The cumulative results suggested that dexamethasone promoted γδ T cell migration out of peripheral blood via an L-selectin-independent mechanism and that dexamethasone did not alter αβ T cell migration kinetics.
Myocardial protection by N,N,N-trimethylsphingosine in ischemia reperfusion injury is mediated by inhibition of P-selectin
Authors: Scalia Rosario , Murohara Toyoaki , Delyani John A , Nossuli Tareck O , Lefer Allan M 
Published: January 2018

Abstract
Polymorphonuclear leukocytes (PMNs) play an important role in myocardial ischemia/reperfusion (MI/R) injury. We examined the cardioprotective effects of N,N,N-trimethylsphingosine (TMS) in a murine model of MI (20 min) and R (24 h) injury in vivo, focusing on leukocyte-endothelial interactions. TMS is a synthetic N-methylated sphingosine derivative that has protein kinase C inhibitory activity and has been shown to prevent leukocyte activation. TMS (18 μg/kg), administered intravenously 1 min prior to reperfusion, significantly attenuated myocardial necrotic injury assessed by myocardial creatine kinase loss compared with MI/R rats receiving only vehicle (P&lt; 0.001). Cardiac myeloperoxidase activity, an index of PMN accumulation in the ischemic myocardium, was also significantly attenuated by TMS compared with rats receiving vehicle (P&lt; 0.001). We further examined whether TMS can attenuate leukocyte-endothelial interaction by intravital microscopy. TMS significantly attenuated NG-nitro-l-arginine–methyl ester (l-NAME)–stimulated PMN rolling and adherence to the rat microvascular endothelium. This action of TMS appears to be mediated by reduction of P-selectin expression because immunohistochemical analysis demonstrated that TMS significantly attenuated endothelial P-selectin expression in the l-NAME-superfused rat mesenteric microvasculature. Similarly, TMS markedly attenuated rapid P-selectin expression in rat platelets stimulated with either thrombin or l-NAME assessed by flow cytometry. In conclusion, TMS seems to be an effective cardioprotective agent by inhibiting early leukocyte-endothelial interaction, thus preventing leukocyte accumulation in the ischemic reperfused myocardium.
Macrophage dysfunction following the phagocytosis of IgG-coated erythrocytes: production of lipid peroxidation products
Authors: Loegering Daniel J , Raley Michael J , Reho Thomas A , Eaton John W 
Published: January 2018

Abstract
The phagocytosis of erythrocytes may contribute to the increased susceptibility to life-threatening infections in patients with burn injury, sickle cell anemia, and malaria. The phagocytosis of Immunoglobulin G–coated erythrocytes (EIgG) is followed by a transient depression of several macrophage functions including phagocytosis, respiratory burst capacity, and killing of bacteria. The present study suggests the possibility that after erythrophagocytosis hemoglobin-derived iron conspires with reactive oxygen products of the macrophage respiratory burst to cause oxidant damage to the phagocyte. Challenge of elicited peritoneal macrophages with EIgG phagocytosis was followed by an increase in lipid peroxidation as assessed by thiobarbituric acid–reactive substances (TBARS). Doses of EIgG associated with increased TBARS also caused a depression of Fc receptor–mediated phagocytosis and phorbol myristate acetate (PMA)–stimulated hydrogen peroxide production. Time course experiments demonstrated that the increase in TBARS coincided with the depression of macrophage function. There was no increase in TBARS following the phagocytosis of IgG-coated erythrocyte ghosts, suggesting that hemoglobin iron is involved in the generation of TBARS. The phagocytosis of erythrocyte ghosts did not depress macrophage function. Since complement receptor–mediated phagocytosis does not stimulate the respiratory burst, the role of the respiratory burst in causing lipid peroxidation was assessed using the phagocytosis of complement-coated erythrocytes. Phagocytic challenge with complement-coated erythrocytes caused neither an increase in TBARS nor a depression of macrophage function. However, there was an increase in TBARS when the respiratory burst was stimulated with PMA following complement receptor–mediated phagocytosis of erythrocytes. These results suggest that hemoglobin iron and phagocyte-generated oxidants collaborate to cause the depression of macrophage function following EIgG phagocytosis.
Respiratory syncytial virus G-protein modulates cytokine release from human peripheral blood mononuclear cells
Authors: König B , Streckert H-J , Krusat T , König W 
Published: January 2018

Abstract
Respiratory syncytial virus (RSV) is the major cause of lower respiratory tract infection in children and adults. In vivo the host has to cope with intact replicative virus, with non-replicative virus, and/or with viral structural proteins including the outer membrane G-protein. We analyzed the role of purified RSV G-protein with regard to its modulatory efficacy for interleukin (IL) -10, IL-12, and tumor necrosis factor-α (TNF-α) release from human peripheral blood mononuclear cells (PBMC). These cytokines seem to contribute to the deleterious effect in viral infections. Treatment of PBMC with RSV at a multiplicity of infection of 10 down to 0.001 induced the release of TNF-α, IL-10, and IL-12; also time kinetics and dose-responses differed markedly. Stimulation of PBMC with purified RSV G-protein (from 0.001 up to 10 μg/106 PBMC) led only to a pronounced increase in IL-10 within a concentration range from 0.01 up to 0.5 μg/106 PBMC with a maximum between 12 and 18 h of incubation. At later time points (24, 48, and 72 h) G-protein concentrations above 1 μg/106 PBMC suppressed IL-10, TNF-α, and IL-12 release from human PBMC. Coating of PBMC with RSV G-protein suppressed IL-10, TNF-α, and IL-12 release after subsequent stimulation with RSV. Our data indicate a regulatory role of RSV G-protein for immune responses toward viral infection.
IL-8 induces T cell Chemotaxis, suppresses IL-4, and up-regulates IL-8 production by CD4+ T cells
Authors: Gesser Borbala , Lund Marianne , Lohse Nicolai , Vestergaard Christian , Matsushima Kouji , Sindet-Pedersen Steen , Jensen Steen Lindkaer , Thestrup-Pedersen Kristian , Larsen Christian Grønhøj 
Published: January 2018

Abstract
Interleukin-8 (IL-8), a neutrophil-activating cytokine, also activates certain T cell functions such as Chemotaxis. We additionally find (n = 6) that recombinant (rIL-8; 1–100 ng/ml), when added to 24 h culture of human CD4+ T cells, suppressed the spontaneous production of IL-4 (50–85%). Steady state production of IL-4 was typically around 30 pg/ml, determined by use of a solid- phase immunoabsorbant assay. De novo synthesis of IL-4 from CD4+ T cells cultured for 3 days was also evaluated by use of detection of [35S]methionine incorporation, as visualized by autoradiography of 2-D gels, and showed that IL-8 suppressed IL-4 production. This suppression of IL-4 production was confirmed in the cytosol fraction by use of Western blotting. The effect of IL-8 (100 ng/ml) was comparable to that of 10 ng/ml recombinant interferon-γ, both strongly suppressing IL-4 production. The regulatory effect of IL-8 on IL-4 production was also indicated by the fact that addition of a neutralizing monoclonal anti-IL-8 antibody (WS.4) enhanced the spontaneous IL-4 production when added to the cultures of CD4+ T cells, thereby probably inactivating the effect of IL-8 originating from the cutured T cells. Also, we observed that IL-4 mRNA expression was down-regulated when the CD4+ T cells were cultured for 12 h in the presence of 100 ng/ml IL-8. The suppression of IL-4 mRNA expression could be prevented by adding anti-IL-8 (20 μg/ml) or IL-10 (100 ng/ml) 1 h before adding rIL-8. Thus, IL-8 may be an important regulator of CD4+ T cell-derived IL-4, thereby possibly regulating the balance between humoral and cellular T cell-dependent responses.
Progesterone inhibits inducible nitric oxide synthase gene expression and nitric oxide production in murine macrophages
Authors: Miller Lance , Alley Evan W , Murphy William J , Russell Stephen W , Hunt Joan S 
Published: January 2018

Abstract
The purpose of this study was to determine whether the female hormones estradiol-17β (E2) and progesterone (P4) influence inducible nitric oxide synthase (iNOS) and the production of nitric oxide (NO) by interferon-γ(IFN-γ)- and lipopolysaccharide (LPS)-activated mouse macrophages. Treatment with P4 alone caused a time- and dose-dependent inhibition of NO production by macrophage cell lines (RAW 264.7, J774) and mouse bone marrow culture-derived macrophages as assessed by nitrite accumulation. RAW 264.7 cells transiently transfected with an iNOS gene promoter/luciferase reportergene construct that were stimulated with IFN-γ/LPS in the presence of P4 displayed reduced luciferase activity and NO production. Analysis of RAW 264.7 cells by Northern blot hybridization revealed concurrent P4-mediated reduction in iNOS mRNA. These observations suggest that P4-mediated inhibition of NO may be an important gender-based difference within females and males that relates to macrophage-mediated host defense.
Neutrophil oxidative metabolism and killing of P. brasiliensis after air pouch infection of susceptible and resistant mice
Authors: Meloni-Bruneri Lucia Helena , Campa Ana , Abdalla Dulcineia S P , Calich Vera Lucia Garcia , Lenzi Henrique L , Burger Eva 
Published: January 2018

Abstract
The oxidative burst of polymorphonuclear neutrophils (PMN) and their ability to inhibit Paracoccidioides brasiliensis growth was studied in susceptible (B10.A) and resistant (A/J) mice. The cells were obtained after subcutaneous inoculation in air pouches, yielding highly pure PMN preparations; the number of cells was similar for both strains at 24 h and five times higher in the resistant strain at 15 days. The oxidative metabolism of these PMN was evaluated by the luminol and lucigen-enhanced chemiluminescence upon stimulation with PMA or killed P. brasiliensis (Pb). At 24 h of infection PMN from both strains showed similar responses. However, at 15 days a great enhancement of the Pb-stimulated luminol-enhanced chemiluminescence was observed only in PMN from resistant mice. Such increase was markedly inhibited by the addition of catalase. Independent of the mouse strain or time of infection the lucigenin-enhanced chemiluminescence showed the same intensity. The lucigenin-enhanced chemiluminescence of PMN without stimuli from resistant mice did not change with the time of infection, however, after 15 days of infection a significantly lower chemiluminescence was detected with PMN from susceptible mice. At 15 days of infection the PMN from B10.A were unable to kill P. brasiliensis yeast cells in vitro. Because the lucigenin- and luminol-enhanced chemiluminescence detects, respectively, the O2- production and the myeloperoxidase/hydrogen peroxide halide system, the present data show parallels between deficiency in the production of oxygen-reactive species by PMN and lower fungicidal activity.
The secretion of preformed granules by macrophages and neutrophils
Authors: Tapper Hans 
Published: January 2018

Abstract
The ability of macrophages and neutrophils to defend tissue homeostasis and participate in inflammatory responses depends on their ability to mobilize granule-membrane proteins and granule content into their external milieu and into phagosomes by regulated secretory processes. Many laboratories have invested much time and effort into furthering our understanding of vesicular transport and secretion. A surge of interest in phagocytosis and phagosomal maturation is also apparent (e.g., the March 1995 issue of Trends in Cell Biology was entirely devoted to phagocytosis). The signaling and the regulation of the secretory response are most likely different for secretion into phagosomes than for secretion into the external milieu. However, these differentially targeted secretory processes rely both upon proteins in vesicular membranes, plasma membrane/phagosomal membrane, and cytosol and upon their interactions with cytoskeletal structures. It is the complex molecular interactions between these components that form the basis for regulation and control of secretion. In the following, the signaling role of granular and cytosolic pH in phagocyte lysosomal secretion is discussed and the current literature on regulated secretion by macrophages and neutrophils is reviewed.
Interleukin-12 protects thermally injured mice from herpes simplex virus type 1 infection
Authors: Matsyo Ryuichi , Kobayashi Makiko , Herndon David N , Pollard Richard B , Suzuki Fujio 
Published: January 2018

Abstract
Severe burn injury is associated with increased susceptibility to severe herpesvirus infections. Type 2 cytokines [interleukin (IL)-4 and IL-10] released from burn-associated CD8+ type 2 T cells (BA-type 2 T cells) have been shown to play a role in the increased susceptibility of thermally injured mice (TI-mice) to herpes simplex virus type 1 (HSV-1) infection. Because IL-12 has been shown to inhibit the generation of type 2 T cells, murine rIL-12 was injected into TI-mice exposed to HSV-1 to determine whether IL-12 could influence HSV-1 infections in individuals bearing type 2 T cells. rIL-12 improved the resistance of TI-mice or mice inoculated with T6S cells (a BA-type 2 T cell clone) against HSV-1 infection. Type 2 cytokines were detected in sera of TI-mice or mice inoculated with T6S cells (T6S-mice). However, treatment of TI-mice or T6S-mice with rIL-12 inhibited type 2 cytokine production in the sera of these mice. All TI-mice exposed to a lethal dose of HSV-1 survived when they were treated with a mixture of monoclonal antibodies (mAbs) against type 2 cytokines. Staphylococcal enterotoxin A [an interferon-γ (IFN-γ) inducer] stimulated serum IFN-γ production in TI-mice and T6S-mice treated with rIL-12, whereas no IFN-γ was produced in mice treated with saline. These results suggest that IL-12 has the potential to protect TI-mice infected with a lethal dose of HSV-1 via a shift to type 1 T cell responses from type 2 T cell responses.