bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interaction of Immune and Connective Tissue Cells: I. The Effect of Lymphokines and Monokines on Fibroblast Growth
Authors: Thornton Stephen C , Por Suzanne B , Walsh Bradley J , Penny Ronald , Breit Samuel N 
Published: January 2018

Abstract
In order to determine if mononuclear cells may be secreting factors capable of modulating fibroblast growth, the in vitro proliferative response of fibroblasts to cytokines known to be secreted by mononuclear cells was measured, using both growth arrested and proliferating cells. Of the cytokines tested, which included interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-3 (IL-3), interleukin-4 (IL-4), interleukin-6 (IL-6), interferon-alpha (IFN-alpha), interferon-gamma (IFN-gamma), transforming growth factor-alpha (TGF-alpha), transforming growth factor-beta (TGF-beta), platelet derived growth factor (PDGF), and tumor necrosis factor-alpha (TNF-alpha), only TNF-alpha and PDGF had demonstrable growth factor activity. Neither IL-1 alpha nor beta showed any true growth factor activity but were able to enhance the replication of already proliferating cells. No inhibition of proliferation was noted by any of the cytokines with the exception of TNF-alpha and TGF-beta. TNF-alpha in doses greater than 500 ng/ml caused fibroblast death, probably by a prostaglandin related mechanism as fibroblasts remained viable, although non proliferative, when assayed in the presence of indomethacin, a known inhibitor of prostaglandin E2 (PGE2) synthesis. TGF-beta was inhibitory to proliferation at doses greater than 100 ng/ml, while fibroblasts remained viable. This effect was not influenced by indomethacin and hence is unlikely to be PGE2 related.
The Effects of 16,16-Dimethyl PGE2 and Phosphodiesterase Inhibitors on Con A Blastogenic Responses and NK Cytotoxic Activity of Mouse Spleen Cells
Authors: Ellis N K , Duffie G P , Young M R , Wepsic H T 
Published: January 2018

Abstract
Prostaglandins function as down regulators of immune responses probably by increasing the concentration of intracellular cAMP. Phosphodiesterase inhibitors, which prevent the breakdown of cAMP, also increase the intracellular levels of cAMP. Prostaglandins and phosphodiesterase inhibitors have both been shown to suppress immune responses in vitro. In this study 16,16-dimethyl PGE2 (dm-PGE2), added in vitro, suppressed the mouse spleen cell concanavalin A (Con A) response by 38% and natural killer (NK) activity by 53%. Addition of the phosphodiesterase inhibitors, theophylline, RO20-1724, or dipyridamole, decreased both the Con A response and NK activity by at least an additional 30%. We also demonstrate that treatment with dm-PGE2 and theophylline in vivo is more effective than either compound alone in inhibiting NK activity of both untreated mice and mice treated with polyinosinic-polycytidylic acid. These studies support the hypothesis that the immunosuppressive effect of dm-PGE2 is mediated by cAMP and suggest that treatment with a combination of dm-PGE2 and phosphodiesterase inhibitors can augment this immunosuppressive effect.
Comparison of Stimulation by Chemotactic Formyl Peptide Analogs Between GTPase Activity in Neutrophil Plasma Membranes and Granule Enzyme Release From Intact Neutrophils
Authors: Kanaho Yasunori , Kermode John C , Becker Elmer L 
Published: January 2018

Abstract
Stimulation by fMet-Leu-Phe analogs of GTPase activity in plasma membranes from rabbit neutrophils was compared with the stimulation of degranulation in intact neutrophils. All four formyl peptides examined (fMet-Leu-Phe-Phe, fMet-Leu-Phe, fNle-Leu-Phe, and fVal-Leu-Phe) were full agonists for both responses. Their ED50 values for the two responses correlated well, although those for GTPase stimulation were uniformly about tenfold greater. The specific antagonist tBoc-Phe-Leu-Phe-Leu-Phe competitively inhibited both GTPase activity and degranulation stimulated by fMet-Leu-Phe; its K values were similar for the two responses. Pertussis toxin treatment, in contrast, inhibited the maximal stimulation of both responses by fMet-Leu-Phe with minimal shift in ED50. The inhibitory actions of tBoc-Phe-Leu-Phe-Leu-Phe and pertussis toxin on GTPase activity thus paralleled the effects on degranulation. These observations substantiate the hypothesis that a guanine nucleotide-binding protein that is a pertussis toxin substrate couples the formyl peptide receptors to physiological function in the neutrophil.
Morphological and Quantitative Aspects of the Development of Cytoplasmic Granules in Concanavalin A-Stimulated Normal Human Lymphocytes
Authors: McGirl Anne , Shea Giles , Melinn Mary , McLaughlin Hugh 
Published: January 2018

Abstract
We have studied normal human lymphocytes stimulated by concanavalin A to determine the rate and extent of development of cytoplasmic granules. Morphological studies on stimulated lymphocytes were undertaken at the light and electron microscopic level. The percentage of T lymphocytes bearing different CD antigens (CD3, CD4, and CD8) that also contained cytoplasmic granules was also assessed. Granules were identified in a high percentage of lymphocytes bearing each of the three CD markers studied.
Activated Macrophages Use Different Cytolytic Mechanisms to Lyse a Virally Infected or a Tumor Target
Authors: LeBlanc Paul A , Heath Lucie S , Um Hong-Duck 
Published: January 2018

Abstract
Murine bone-marrow-culture-derived-macrophages can be differentially activated to lyse either vesicular stomatitis virus infected BALB/c3T3 cells or the tumor target P815. Macrophages were activated in a manner so that they could lyse both targets. The ability of this activated population to lyse either target type was differentially inhibited by varying the assay conditions. The lysis of P815 targets was more sensitive to inhibition by the proteinase inhibitor N-p-tosyl-L-lysine chloromethyl ketone than was the lysis of virally Infected cells. On the other hand, reduction of the concentration of glucose in the assay medium, which inhibits the production of oxygen metabolytes by the hexose monophosphate shunt, or the addition of anti-tumor necrosis factor (anti-TNF) serum were able to decrease the lysis of virally infected targets but not P815 targets. Thus, the observed differences in the lysis of these two targets were due to both the activation state of the macrophages and the differential susceptibility of the targets to different effector mechanisms.
Identification of the Major Glycosyl–Phosphatidylinositol Anchored Proteins on the Surface of Human Neutrophils
Authors: Skubitz Keith M , Lakatua John D A 
Published: January 2018

Abstract
A novel mechanism of protein attachment to cell membranes involving the covalent linkage of the protein through an oligosaccharide to phosphatidylinositol has recently been defined. Many proteins that are anchored to the cell membrane by this mechanism can be released by treatment with phosphotidylinositol-specific phosphollpase C (PI-PLC). Monoclonal antibodies are useful as probes in the study of the roles of cell-surface components in neutrophil function. Many monoclonal antibodies that bind to human neutrophils react with the oligosaccharide lacto-N-fucopentaose III (CD15 antibodies). Human neutrophil surface proteins identified by 125l surface-labeling using lactoperoxidase were examined for PI-PLC sensitivity, to identify the major surface proteins of human neutrophils that are anchored by a glycosyl–phosphatidylinositol linkage. The major surface-labeled protein identified by lactoperoxidase-catalyzed iodination was a ~68–90-kDa protein. Three major surface proteins identified by 125I-surface labeling of ~68–80, 57, and 33-kDa were released by PI-PLC treatment Immunoprecipitation and subsequent gel electrophoresis of proteins from neutrophils labeled with 125l revealed a previously unidentified 98–115-kDa protein specifically reactive with CD15 antibodies that was released from the cell by treatment with PI-PLC. The roles of these proteins in neutrophil function remain to be determined.
Activation of the Superoxide-Generating NADPH Oxidase of Macrophages by Sodium Dodecyl Sulfate in a Soluble Cell-Free System: Evidence for Involvement of a G Protein
Authors: Aharoni Iris , Pick Edgar 
Published: January 2018

Abstract
The superoxide (O2-)-generating NADPH oxidase of resting macrophages can be activated in a soluble cell-free system by certain anionic amphiphiles, such as sodium dodecyl sulfate (SDS). We demonstrate that cell-free activation is specifically enhanced by nonhydrolyzable guanosine 5′-triphosphate (GTP) analogues. Guanosine 5′-diphosphate (GDP) and guanosine 5′-O-(2-thiodiphosphate) (GDPβS) prevent cell-free oxidase activation and reverse the activated state when added to preactivated oxidase preparations. Nonhydrolyzable GTP analogues have a protective effect on the SDS-stimulated NADPH oxidase, as shown by the maintenance of more than 90% and close to 60% of enzyme activity 6 and 24 hr, respectively, after the addition of SDS to the cell-free preparation. A novel procedure is described for separating the activated NADPH oxidase from SDS and added nucleotides by gel filtration of the SDS-stimulated solubilized membrane-cytosol mixtures through a Sephadex G-25 column. By utilizing this method, it was found that the presence of micromolar concentrations of nonhydrolyzable GTP analogues during activation by SDS results in a marked increase in the recovery of SDS-independent, NADPH-dependent O2--producing activity in the excluded volume of the column. It is suggested that GTP stabilizes the SDS-induced complex between membrane and cytosolic components of the oxidase. Cell-free activation of NADPH oxidase by SDS was found to be cholera and pertussis toxin insensitive. These results serve as evidence of the participation of a G protein in the activation of the O2--generating NADPH oxidase of macrophages by SDS.
Neutrophil Accumulation and Plasma Leakage Induced In Vivo by Neutrophil-Activating Peptide-1
Authors: Colditz I G , Zwahlen R D , Baggiolini M 
Published: January 2018

Abstract
Neutrophil accumulation and plasma leakage induced in rabbit skin by neutrophil-activating peptide-1 (NAP-1, a 72 amino acid peptide produced by monocytes and a variety of tissue cells), E. coli endotoxin, and interleukin-1 (IL-1) were compared. Neutrophil accumulation at sites injected with NAP-1 was intense, rapid, and long-lasting; it reached a maximum rate during the first 30 min, continued at constant rate for 4–6 h, and remained detectable up to at least 8 h. In contrast, the neutrophil-attracting effect of endotoxin and IL-1 was slower in onset and more transient; it peaked in the first 2 h and declined to a very low level after 4 h. Plasma leakage induced by NAP-1 had a shorter time course than neutrophil accumulation and ceased after 6 h. Depletion of blood neutrophils by treatment with hydroxyurea prevented the plasma leakage induced by NAP-1 or endotoxin but not by histamine. Desensitization to NAP-1 was studied by restimulation of lesions. Following restimulation with NAP-1 after intervals from 6–10 h, there was diminished infiltration of neutrophils, while nearly normal responses were obtained after an interval of 24 h. Desensitization was dose dependent and affected both plasma leakage and neutrophil accumulation. In lesions initiated with NAP-1 there were normal responses following restimulation with endotoxin but marked desensitization to IL-1, suggesting that NAP-1 may contribute to inflammation induced by IL-1 but not by endotoxin.
This study indicates that NAP-1 is a potent mediator of neutrophil accumulation in vivo, with characteristics similar to those reported for C5 fragments, but with a more protracted action.
Transient Expression of Virus-Specific Promoters in Murine Resident Peritoneal Macrophages
Authors: Wu Linxian , Anaraki Farvardin , Morahan Page S , Leary Kathryn 
Published: January 2018

Abstract
Monocyte-macrophages (MO), being non-permissive for most viruses, play an important role in resistance to virus infection. In order to establish the mechanism of abortive infection of murine resident peritoneal MO (ResPMO) by herpes simplex virus type 1 (HSV-1), it is desirable to transfect these cells with viral promoters linked to an assayable gene, for example, the bacterial chloramphenicol acetyl transferase (CAT) gene. This will facilitate studies designed to measure levels of promoter activation or repression in these specialized cells. Transient expression of CAT in ResPMO was achieved with DEAE-dextran, but not using either calcium phosphate precipitate or lipofectin. CAT expression driven by various virus-specific promoters was less efficient in ResPMO compared with Vero cells and approximately 50% of input plasmid DNA remained in Vero cells at 48 h post transfection, but only 9% was detectable in ResPMO. However, approximately 6% of ResPMO and 9% of Vero cells contained CAT-specific DNA at 24 h post transfection. In addition, 2% of cells of either cell type contained CAT-specific polypeptide at 48 h. This is therefore the first report that the non-replicating murine ResPMO can be transfected in vitro and more importantly, that these cells express the transfected gene products.
Lysis of Autologous Human Macrophages by Lymphokine-Activated Killer Cells: Interaction of Effector Cell and Target Cell Conjugates Analyzed by Scanning Electron Microscopy
Authors: Streck Richard J , Helinski Ernest H , Ovak Geraldine M , Pauly John L 
Published: January 2018

Abstract
Lymphokine (i.e., interleukin 2; IL-2)-activated killer (LAK) cells derived from normal human blood are known to destroy human tumor target cells. Accordingly, immunotherapy modalities using IL-2, either alone or in combination with LAK cells, have been evaluated for eradicating metastatic cancer. In studies conducted to characterize receptors on LAK cell membrane ultrastructures, we observed that LAK cells kill autologous human monocyte-derived macrophages (Mφ). In these experiments, peripheral blood mononuclear cells of a healthy adult donor were cultured to generate LAK cells and autologous nonadherent Mφ. Thereafter, conjugates were prepared by incubating for 3 h autologous populations of LAK cells and Mφ. Examination of the conjugates by scanning electron microscopy (SEM) identified LAK cell-mediated killing of Mφ. Moreover, SEM analysis of the LAK cell membrane architecture identified microvilli-like ultrastructures that provided a physical bridge that Joined together the LAK cell and Mφ. The immunological mechanism(s) underling LAK cell killing of autologous Mφ is not known; nevertheless, these conjugates will provide a useful model to study membrane receptors on ultrastructures that mediate the initial stages of cytolysis that include target cell recognition and cell-to-cell adhesion. The results of our observations and the findings of other investigators who have also demonstrated LAK cell killing of autologous normal human leukocytes are discussed in the context of the association of IL-2 and IL-2-activated killer cells with side effects observed in ongoing clinical trials and with autoimmune disorders.
Production of Collagen Type I by Mouse Peritoneal Macrophages
Authors: Vaage Jan , Lindblad William J 
Published: January 2018

Abstract
The origin and identity of the cells that accumulate and produce collagen in wound healing and in granulomatous and fibrotic processes have long remained unsettled. In this study, it was found that pure cultures of peritoneal macrophages produced hydroxy-L-proline not associated with Clq and produced detectable quantities of collagenase-digestible protein. Cultured macrophages synthesized collagen while retaining their phagocytic capacity. Immuno-blot analysis identified the collagen as Type I. This shows that macrophages have collagen-forming capacity and may be productive cells in pathologic collagen deposition.
Computer Image Analysis Method for Rapid Quantitation of Macrophage Phagocytosis
Authors: Odeyale Charles O , Hook Gregory R 
Published: January 2018

Abstract
A new method is described for rapidly quantitating phagocytosis by adherent macrophages in culture using computer image analysis (CIA) of video light microscopic images. Ingestion of fluorescent microspheres by peritoneal murine macrophages is used to model phagocytosis. The grey levels of digital phase contrast and fluorescent microscopic images are used to quantitate the number of microspheres per cell. The method is semi-automatic, analyzes approximately 2 × 103 cells/hr, and simultaneously measures phagocytosis (microspheres/cell), cell area, and density (number of cells/mm2). CIA obtains the same microspheres/cell average as manual microscopic counting and an analytical precision of 5%. As expected, CIA found that the number of microspheres/cell linearly increases with increasing macrophage-microsphere co-culture time or increasing microsphere concentration until macrophages become saturated. CIA finds increased phagocytosis by interferon-gamma-treated cells and suppressed phagocytosis by cytochalasin B- or 4°-treated cells relative to controls, which demonstrates that CIA can resolve biological changes in macrophage phagocytosis. CIA also provides quantitative data on macrophage morphometry and density and found an increase in the cell area and density of INF treated macrophages. CIA provides significantly more phagocytic, morphometric, and density data than conventional manual microscopic counting methods or flow cytometric methods. The limitations, improvements, and future applications of this method are discussed.
Role of Adherence vs. Spreading in the Induction of Membrane-Associated Interleukin-1 on Mouse Peritoneal Macrophages
Authors: Labadia Mark , Faanes Ronald B , Rothlein Robert 
Published: January 2018

Abstract
Adherence of peritoneal exudate cells (PEC) to plastic has been shown to activate several PEC functions, such as tumor ceil lysis and membrane-associated interleukin-1 (mlL-1) expression. Several studies have demonstrated that leukocyte adherence is dependent on divalent cations. In this study, ethylenediaminetetrascetic acid (EDTA), a known chelator of divalent cations, was used to evaluate the role of cell attachment vs. spreading in adherence-induced mlL-1 activity on resident C57BL/6 mouse PEC. Significant inhibition of PEC spreading on plastic and mlL-1 expression was noted when PEC were cultured in the presence of 10 mM EDTA. However, PEC remained adherent in the presence of EDTA and were able to express mlL-1 activity in response to a soluble stimulus lipopolysaccharide (LPS) 1 μg/ml. These results suggest that the divalent cation-dependent spreading of PEC on plastic initiates or enhances the expression of mlL-1 activity. Additionally, adhesion and LPS stimulate mlL-1 expression by independent mechanisms.
Role of Interferon-γ in Counteracting the Suppressive Effects of Transforming Growth Factor-β2 and Glucocorticoids on the Production of Tumor Necrosis Factor-α
Authors: Dunham Diane M , Arkins Sean , Edwards Carl K , Dantzer Robert , Kelley Keith W 
Published: January 2018

Abstract
The multipotential cytokine, transforming growth factor-β2 (TGF-β2), is as effective as glucocorticoids in suppressing the production of tumor necrosis factor-α (TNF-α) by lipopolysaccharide (LPS)-stimulated macrophages, and this inhibition can be abrogated by exogenous interferon-γ (IFN-γ). Porcine alveolar macrophages triggered with LPS produce TNF-α as identified by complete blocking of cytotoxicity on WEHI 164 clone 13 cells in macrophage supernatants by a monoclonal antibody to human TNF-α. Platelet-derived porcine TGF-β2, at a concentration of 4 nM, inhibited LPS-induced production of TNF-α by 93%. Dexamethasone was as effective as TGF-β2, suppressing TNF-α production by 86% at a concentration of 4 nM. The natural but less potent glucocorticoid Cortisol inhibited TNF-α production by 100% at a 100-fold higher concentration (400 nM). Recombinant PoIFN-γ consistently primed LPS-triggered macrophages for increased production of TNF-α by 50-100%, and this priming was totally blocked by a polyclonal antibody to rPoIN-γ. Furthermore, the suppression in LPS-induced production of TNF-α caused by TGF-β2, dexamethasone, and Cortisol could be reversed by addition of rPoIFN-γ. These data show that alveolar macrophages can be effectively primed by rPolFN-γ even in the presence of moderately suppressive doses of TGF-β2 and antiinflammatory steroids.
Avian Inflammatory Macrophage Function: Shifts in Arachidonic Acid Metabolism, Respiratory Burst, and Cell-Surface Phenotype During the Response to Sephadex
Authors: Golemboski K A , Whelan J , Shaw S , Kinsella J E , Dietert R R 
Published: January 2018

Abstract
The avian inflammatory response to intraperitoneal (i.p.) Sephadex injection produces macrophages which display characteristics of an increasingly activated state over time. We examined elicited chicken peritoneal exudate cells (PECs) with respect to superoxide anion production, arachidonic acid metabolism and cell surface la and transferrin receptor (TfR) expression from 4 to 96 h after i.p. stimulation. Avian PECs showed the highest level of superoxide release when harvested just 4 h after injection, and did not produce PGE2 or 6-keto PGF1α. Early (4-h) PECs produced elevated amounts of thromboxane as compared to later (42-h) macrophages. Expression of both la and TfR increased between 4 and 24 h after Sephadex stimulation; TfR remained elevated through 96 h, but la declined after 42 h. Some aspects of chicken macrophage regulation of superoxide anion, thromboxane release, and surface antigen expression are in contrast with those reported for mouse macrophages.
Lectin Binding of Rat Bone Marrow Cells During Colony-Stimulating Factor Type 1–Induced Differentiation: Soybean Agglutinin as a Marker of Mature Rat Macrophages
Authors: Krugluger Walter , Gessl Alois , Boltz-Nitulescu George , Förster Othmar 
Published: January 2018

Abstract
Rat bone marrow cells (BMC) cultured in the presence of murine colony-stimulating factor type 1 (CSF-1) differentiate within 7 days into a cell population containing 96–100% macrophages (Mφ). In this study, binding of 10 different fluorescein isothiocyanate (FITC)-conjugated lectins to cultured BMC at various stages of differentiation into Mφ was investigated. Only soybean agglutinin (SBA) showed a binding pattern that was significantly correlated to Mφ differentiation. Nearly all adherent (A) cells bound SBA. Binding of SBA to nonadherent (NA) cells increased from 20% on day 0 to 80% on day 8. NA cells were separated by means of a fluorescence-activated cell sorter into SBA-, SBA±, and SBA+ fractions. On day 0 and day 2, Mφ, blast-like cells, eosinophils, and some neutrophils were found in the SBA+ population. From day 4 onwards, the SBA+ fraction contained almost exclusively Mφ. Neutrophils and some blasts were found in the SBA-population on days 0 and 2. In the SBA± fraction, mainly blasts and lymphocytes were identified. With increasing time of culture, Mφ or Mφ precursors prevailed also in the SBA-/± cell population. Cells forming colonies in soft agar in the presence of CSF-1 were highly enriched in the SBA± fraction. A large number of cells in S-G2/M phases but few colony-forming cells were found in the SBA+ population. Our data suggest that SBA is a useful additional tool to define late stages of Mφ differentiation.