bluuebunny/crossref_metadata_2025_split huggingface dataset:
Macrophage-Like Tumor Cells as Tools to Study Chemoattractive Activity
Authors: Terheggen P , Van Loveren H , Den Otter W 
Published: January 2018

Abstract
Macrophage-like tumor cells can be obtained in large quantities as rather homogeneous populations, making these cells useful for chemotaxis assays. Therefore, macrophage-like cells J774A, WEHI-3, P388D1, IC-21, and NCTC 1469, all of murine origin, and U937 of human origin, were tested for chemotactic activity to a number of chemoattractive agents, such as casein, an N-formyl tetrapeptide (N-formyl-L-norleucyl-L-leucyl-L-phenylalanyl-L-tyrosine), and culture supernatants of murine SL2 lymphoma cells. J774A and WEHI-3 macrophage-like cells of murine (BALB/c) origin expressed the strongest chemotactic activity to casein and N-formyl tetrapeptide, respectively. The results show that: very standardized chemotaxis assays can be performed using these cell lines; these assays require appropriate cell line–stimulus combinations; there are substantial differences among cell lines as to sensitivity to various chemoattractive substances; macrophage cell lines and functional mutants may be helpful for the study of receptors for chemotaxins and the study of transducer signals for chemotaxis.
Effect of Alkylating Agents on the Clearance of Staphylococcus aureus From Murine Lungs
Authors: Nugent Kenneth M , Onofrio John M 
Published: January 2018

Abstract
Alkylating agents have several effects on cellular host defense responses which could increase either the frequency or the severity of pulmonary infections. In addition, some of these agents directly injure lung parenchyma and could have effects on intrapulmonary killing processes independent of any effect on phagocyte number and function. We have used a murine model for staphylococcal clearance to evaluate the effect of cyclophosphamide and mechlorethamine on intrinsic lung defenses. Single doses of mechlorethamlne (40 μg IV) or of cyclophosphamide (150 mg/kg IP) reduce peripheral blood neutrophil counts and spleen weights on day 3 after injection; with the exception of neutrophil counts in mechlorethamine-treated mice, these parameters returned to normal by days 10-12. Both drugs reduced the number of alveolar macrophages recoverable by bronchoalveolar lavage on days 10-12 but not day 3. Mechlorethamine delayed the clearance of Staphylococcus aureus 502A from the lung on both days 3 and 12, but cyclophosphamide did not alter clearance on either day 3 or 10. The defect in clearance in mechlorenthamine-treated mice resolved by 3 wk after drug administration. These results demonstrate that alkylating agents do not have uniform effects on antibacterial processes in the murine lung. Since the mechlorethamine effect on staphylococcal elimination appears independent of its effect on macrophage numbers, these results suggest that staphylococcal clearance also depends on nonphagocytic host defense factors.
The Kinetics of Secretion From Permeabilized Human Neutrophils: Release of Elastase and Correlations With Other: Granule Constituents and Right Angle Light Scatter
Authors: Smolen James E , Stoehr Sally Jo , Traynor Alexis E , Sklar Larry A 
Published: January 2018

Abstract
We have previously reported that human neutrophils can be permeabilized with the cholesterol-complexing agent digitonin. These permeabilized cells can be induced to secrete lysosomal constituents when exposed to micromolar levels of free Ca2+, a process that is enhanced by certain guanine nucleotides. We examined the kinetics in this system by employing both direct and indirect measures of secretion. A continuous, fluorescent assay of elastase permits real-time monitoring of secretion from azurophil granules. The kinetics of elastase release proved to be rapid, beginning within 3-10 sec and reaching a maximum at 1-2 min. Changes in the Ca2+ concentration did not affect the “lag period” for release. A comparison of the Ca2+ dose-response curves for release of the various granule constituents indicated that elastase was being secreted along with other contents of the azurophil granules. Changes in right angle light scatter (RLS), which have been shown to correlate closely with secretion, also commenced rapidly after the addition of Ca2+; when measured simultaneously, both the Ca2* dose-response characteristics for changes in RLS and elastase release were very similar. Changes in RLS could be halted within 5 sec by excess EGTA and restarted promptly by repletion with secretory concentrations of Ca2+. In addition, neomycin, a phospholipase C inhibitor, profoundly diminished degranulation as monitored by RLS and end-point techniques. A continuous assay employing 9-aminoacridine self-quenching as a measure of secretion proved far less satisfactory, but, nonetheless, produced similar kinetics and dose-response characteristics. These data thus suggest that secretion of granule constituents from permeabilized neutrophils takes place rapidly and that the kinetics are comparable to those observed with intact cells.
Candidacidal Activity of Monocyte-Derived Human Macrophages: Relationship Between Candida Killing and Oxygen Radical Generation by Human Macrophages
Authors: Sasada Masataka , Kubo Akemi , Nishimura Toshiro , Kakita Tokio , Moriguchi Toshinori , Yamamoto Kohkichi , Uchino Haruto 
Published: January 2018

Abstract
Freshly isolated human monocytes ingested and killed Candida albicans, and generated O-2 H2O2 and .OH efficiently. When monocytes were cultured in vitro, these cells transformed into macrophages. Cultured monocytes retained their ingestive activity but lost their candidacidal activity almost completely after day 3. The release of O-2 by monocytes decreased slightly with culture and that of .OH was markedly decreased on day 3 of culture. The activity of myeloperoxidase in the monocytes decreased with culture. These results suggested that the loss of candidacidal activity is due to the decrease of .OH generation and myeloperoxidase activity in cultured monocytes.
Receptor-Mediated Ingestion Responses by Lung Macrophages From a Canine Model of ARDS
Authors: Tabor Dale R , Kiel Deborah P , Jacobs Richard F 
Published: January 2018

Abstract
Receptor-mediated ingestion was examined in macrophages derived from a canine model of the adult respiratory distress syndrome (ARDS). The results showed that Fc-mediated ingestion by alveolar macrophages (AM) and macrophages from lung parenchyma (PM) was significantly diminished when compared with their respective controls. Pulsing all the experimental groups with lipopolysaccharide (LPS) for 1 hr in vitro failed to either enhance the response or return the activity to levels achieved by control cells. In parallel studies, an analysis of C3b-mediated ingestion showed that both the experimental AM and PM performed this function only at a magnitude equal to the control cells. Similar responses were observed when an LPS pulse was performed. Although there was a reduction in Fc-mediated ingestion and an apparent restraint of the C3b-mediated ingestion, both AM and PM expressed a significantly enhanced ability to spread. These results suggested that the canine model of ARDS alters at least one select macrophage function that may be important to subsequently protect the host. Such disturbances in the cellular immune response may contribute to the progression of infection and lung pathology associated with this disease process.
A High-Yield, High-Purity Elutriation Method for Preparing Human Granulocytes Demonstrating Enhanced Experimental Lifetimes
Authors: Tolley James O , Omann Geneva M , Jesaitis Algirdas J 
Published: January 2018

Abstract
A counterflow centrifugal elutriation procedure for preparing human granulocytes is described. This method permits isolation of 1-2 × 109 granulocytes with a yield of 70% ± 7% from a unit of whole blood in 3 h. These cells are 99% ±1% pure and 95-99% viable. When compared to cells prepared by a conventional procedure employing hypotonic lysis to remove red cells, the elutriated cells show enhanced oxidative responses resulting from enhanced longevity. Other responses including receptor-mediated uptake of chemoattractant were not significantly different from those of conventionally prepared cells after incubation at 37° prior to stimulation. The population of elutriated granulocytes, however, was significantly more homogeneous and pure than cells prepared conventionally.
Transient Suppression of the Zymosan-Induced Chemiluminescence Produced by Bone Marrow Cells During a Subcutaneous Nonspecific Inflammation in the Mouse
Authors: Herodin Francis , Benichou Gilles 
Published: January 2018

Abstract
The oxidative metabolism of remote phagocytes has been studied during the course of an acute nonspecific inflammation in the mouse. The bone marrow cells (BMC) from mice bearing a polyacrylamide-microbead-induced granuloma (Biogels P4 and P100) show a transient striking decrease of their chemiluminescence (CL) response to opsonized zymosan. This decreased oxidative response occurs between the third and 72nd hr, with a minimum observed at the 24th hr. Bone marrow granulocytes are assessed to represent the main chemiluminescent BMC compartment, using complement-mediated cytotoxicity assays with a monoclonal antimouse granulocyte antibody. Biogel P4-treated mice, but not Biogel P100-treated ones, show at the 24th hr a significantly decreased percentage of segmented bone marrow granulocytes (-45%, P &lt; .001).
The accurate mechanisms involved in this transient suppression of stimulus-induced CL produced by BMC remain to be elucidated.
Morphologic and Functional Heterogeneity Among Rat Alveolar Macrophage Fractions Isolated by Centrifugation on Density Gradients
Authors: Oghiso Yoichi 
Published: January 2018

Abstract
Lavaged rat bronchoalveolar cells were separated into four different density fractions (I—IV) by centrifugation through a discontinuous Percoll gradient. Maximum cell size was found in the lowest density fraction (I) and minimal cell size in the highest density fraction (IV), showing an inverse correlation with cell density. These fractions contained alveolar macrophages (AM) in the proportion of 97% or more by morphologic criteria, while phagocytic AM was approximately 80% in each fraction. Although the proportion of la antigen-positive AM was low in each fraction, it was elevated after incubation with supernatants from concanavalin A (Con A) stimulated spleen cell cultures, with greater la expression in higher density fractions. Differences in these fractions were also noted in several functions, including Fc receptor activity, chemotactic migration, tumoricidal activity, and interleukin 1 (IL-1) production, all of which were greater in higher density fractions (III and IV). Con A-induced T cell proliferation was, however, suppressed by these higher density fractions, whereas only an intermediate density fraction (II) enhanced T cell responses. These results indicate morphologic and functional heterogeneity among rat AM.
The Contributions of Adrenal Hormones, Hemodynamic Factors, and the Endotoxin-Related Stress Reaction to Stable Prostaglandin Analog-Induced Peripheral Lymphopenia and Neutrophilia
Authors: Ulich Thomas R , Keys Marcy , Ni Rong-Xiang , del Castillo Juan , Dakay Edwin B 
Published: January 2018

Abstract
Stable prostaglandin analogs are known to induce lymphopenia and neutrophilia in a dose-dependent fashion after subcutaneous injection in rats. The purpose of the present investigation is to determine whether the prostaglandin-induced changes in circulating leukocytes might be secondary to hypotension with the ensuing release of adrenal hormones. The adrenal medullary catecholamine epinephrine was found to induce neutrophilia in both intact and adrenalectomized rats, and the glucocorticosteroid analog dexamethasone induced a profound lymphopenia in rats as reported by previous investigators. A stable analog of PGF2α (15-S-15-methyl PGF2α; M-PGF2α) at the dose of 1 mg/kg induced marked systemic hypotension 1 h after injection, with lymphopenia and neutrophilia 6 h after injection. The non-prostanoid hypotensive agent captopril, at a dose of 63 mg/kg, induced a hypotension of similar magnitude and kinetics to that induced by prostaglandin. Captopril also induced lymphopenia and neutrophilia at 6 h, although the neutrophilia was of lesser magnitude than that induced by prostaglandins. The prostaglandin-induced lymphopenia was found to be mediated, at least in part, by the hypotension-induced release of adrenal hormones, as evidenced by the abrogation of lymphopenia in prostaglandin-treated adrenalectomized rats. Captopril-treated adrenalectomized rats, however, did develop a significant lymphopenia, suggesting that hypotension can result in lymphopenia even in adrenalectomized rats. The M-PGF2α-induced neutrophilia in adrenalectomized rats, by comparison to captopril-induced neutrophilia in adrenalectomized rats, was greater than the neutrophilia expected as the result of hypotension alone. Indeed, the M-PGF2α-induced neutrophilia in adrenalectomized rats was greater than the captopril-induced neutrophilia in sham-adrenalectomized rats. Thus, a portion of the neutrophilia induced by M-PGF2α in intact rats may be mediated through adrenal-independent, hemodynamic-independent mechanisms. The possibility that M-PGF2α might be inducing neutrophilia via an endotoxin-like stress reaction was investigated by examining changes in circulating white blood cells in intact and adrenalectomized C3H/HeN (endotoxin-sensitive) and C3H/HeJ (endotoxin-resistant) mice after prostaglandin administration. No quantitative differences in the prostaglandin-induced neutrophilia were noted in C3H/HeJ mice as compared to the C3H/HeN mice. In conclusion, the effects of M-PGF2α on circulating white blood cells in the intact rat are mediated in part through hemodynamic and adrenal mechanisms, and in part through adrenal-independent, hemodynamic-independent, and endotoxin-like stress reaction-independent mechanisms. Prostaglandins may contribute directly to the genesis of neutrophilia.
Mannose Inhibits the Human Neutrophil Oxidative Burst
Authors: Rest Richard F , Farrell Catherine F , Naids Fredrick L 
Published: January 2018

Abstract
Stimulated human neutrophils (PMNs) increase their oxygen consumption and secrete reactive oxygen species that are involved in bactericidal activity and inflammation. While studying lectin-mediated bacterial adherence, we observed that D-mannose appeared to inhibit PMN metabolism. Further studies showed that 100 mM mannose inhibited oxygen consumption by 82%, superoxide secretion by 84%, luminol-enhanced chemiluminescence (CL) by 98%, and hexose-monophosphate shunt activity by 100% when PMN were stimulated with 1 μM phorbol myristate acetate (PMA). Inhibition was also seen with 0.1 μM formyl-methionyl-leucyl-phenylalanine (fMLP), and 0.1 μM A23187, reagents thought to stimulate the respiratory burst by different transductional mechanisms. Inhibition was dose-responsive and specific since 100 mM D-galactose, alpha-D-glucose, or alpha-L-fucose only minimally affected PMN oxidative metabolism. Inhibition of PMA-induced superoxide production was seen almost immediately upon the addition of 50 mM mannose and was reversed by washing. Neutrophils remained viable as measured by trypan blue exclusion. These data suggest that mannose inhibits the neutrophil oxidative burst at the level of the hexose monophosphate shunt. Further investigation should elucidate the specific mechanism(s) of this burst inhibition as well as define uses for it as a tool to study oxidative as well as nonoxidative killing by PMN.
Enhanced Secretion of Immunoreactive Bombesin by Alveolar Macrophages Exposed to Silica
Authors: Wiedermann C J , Adamson I Y R , Pert C B , Bowden D H 
Published: January 2018

Abstract
Bombesin has recently been identified in alveolar macrophages (AM). Since this peptide has been shown to stimulate fibroblast growth in culture, we wished to determine whether AM exposed to the fibrogenic particle silica in vivo were capable of secreting more bombesin than AM recovered after instilling inert carbon particles to the lung. Rats received 10 mg of either carbon or silica by intratracheal injection and were killed at 3 days or 6 weeks. Both particles induced a rapid inflammatory response, and normal levels of immunoreactive bombesin were measured in lung lavage fluid and in freshly recovered macrophages from all rats. However, incubation of normal AM for 4 h in serum free medium produced a significant increase in bombesin levels measured in supernatants. Bombesin in supernatants of AM cultured after recovery from rats exposed to carbon was at the control value, while AM recovered after silica exposure in vivo secreted increased amounts of bombesin when cultured. Cells recovered 6 wk alter instilling silica to the lung and cultured for 4 h secreted 50% more bombesin than control AM. At this time, hydroxyproline measured in the silica-injected lungs was also significantly higher than in controls or carbon-injected rats. These results indicate that AM recovered from lungs after exposure to silica secrete increased amounts of bombesin during the development of pulmonary fibrosis.
Low-Density Lipoprotein Oxidized by Polymorphonuclear Leukocytes Inhibits Natural Killer Cell Activity
Authors: Tanabe Fuminori , Sato Akihiko , Ito Masahiko , Ishida Eiko , Ogata Masahiro , Shigeta Shiro 
Published: January 2018

Abstract
We studied the effect of low-density lipoprotein (LDL) oxidized by opsonized zymosan-stimulated polymorphonuclear leukocytes (PMN) on natural killer (NK) cell activity. Oxidized LDL inhibited NK cell activity in a dose-dependent manner, whereas normal LDL left it unaffected. However, oxidized LDL did not inhibit antibody-dependent cell-mediated cytotoxicity (ADCC). Moreover, a positive correlation was observed between the amount of thiobarbituric acid-reacting substances (TBARS) on the sample of oxidized LDL and the degree of inhibition of NK cell activity. We also showed that oxidized LDL suppressed the binding capacity of purified large granular lymphocytes (LGL) to target cells without changing the lytic activity. These results therefore suggest that activated PMN can modulate NK cell activity by oxidizing LDL.
Lectin Interactions With the Jurkat Leukemic T-Cell Line: Quantitative Binding Studies and Interleukin-2 Production
Authors: Dupuis Gilles , Bastin Bânû 
Published: January 2018

Abstract
Phytohemagglutinin (PHA), concanavalin A (Con A), pea lectin, and wheat germ agglutinin (WGA) have been used to investigate their binding properties to Jurkat 77 6.8 leukemic human T cells and their ability to induce these cells to produce interieukin-2 (IL-2). Binding studies showed that the Jurkat cells fixed 0.82 ± 0.11 μg pea lectin, 2.02 ± 0.17 μg Con A, 1.85 ± 0.07 μg PHA and 8.88 ± 0.61 μg WGA. Scatchard plots were linear, indicating that the binding process was homogeneous with respect to the binding constant. PHA and Con A bound with the highest affinity [K (apparent) - 9 × 109 M−1], followed by pea lectin and WGA [K, (apparent) =3 × 109 M−1]. The number of lectin binding sites was in agreement with the results of saturation experiments. We also evaluated the effect of the presence of 12-O-tetradecanoylphorbol-13-acetate (TPA) on the binding process. Results show that there were no gross alterations in the value of (apparent) K in the case of PHA and WGA. In contrast, the presence of TPA decreased the affinity of Con A and modified the Scatchard profile for pea lectin, which was curvilinear with a concavity turned upward. In this case, data were (apparent) K1 = 17.7 × 109 M−1 (“high-affinity” sites) and (apparent) K2 = 2.6 × 109 M−1 (“low-affinity” sites). The four lectins shared the ability to stimulate Jurkat 77 6.8 cells to secrete IL-2. Optimal lectin concentrations were 20 μg/ml (PHA) and 50 μg/ml (WGA and Con A). Pea lectin failed to display a dose-response relationship, and IL-2 production increased proportionally with lectin concentration. Con A was the most efficient stimulator (250 U/ml), followed by WGA (160 U/ml) and PHA (108 U/ml). Addition of TPA potentiated Jurkat cell response. In this case, Con A and pea lectin were the most efficient stimulators (470 U/ml), followed by PHA (316 U/ml) and WGA (271 U/ml). Here, also, pea lectin failed to show a dose-response relationship. Our data do not reveal an obvious relationship between the binding parameters and the ability of the lectins to induce IL-2 production. We suggest that the efficiency of the four lectins to stimulate IL-2 production in the case of the Jurkat 77 6.8 variant is most likely related to the nature of their specific cell surface receptor(s).
Effects of Ammonium Metavanadate Treatment Upon Macrophage Glutathione Redox Cycle Activity, Superoxide Production, and Intracellular Glutathione Status
Authors: Cohen M D , Wei C I 
Published: January 2018

Abstract
Female B6C3F1 mice were given intraperitoneal injections of ammonium metavanadate (2.5 or 10 mg V/Kg), ammonium chloride, or sodium phosphate buffer every 3 days for 6 weeks. Resident peritoneal macrophages were harvested, lysed by freeze-thawing, and the resulting cytolysate was assayed for total protein content and enzyme activities of glutathione reductase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. In addition, peritoneal macrophages were assayed for superoxide production using nitroblue tetrazolium reduction, as well as for intracellular levels of oxidized and reduced glutathione. Exposure of mice to vanadium resulted in a dose-trend depression in the three macrophage enzyme activities as compared with the controls. Vanadium treatment resulted in a similar decrease in the production of superoxide anion, and an increase in levels of oxidized glutathione; however, the total glutathione pool (reduced plus oxidized forms) was not affected.
Preparation and Characterization of a Monoclonal Antibody That Inhibits Human Neutrophil Elastase Activity
Authors: Skubitz Keith M , Wehner Nancy G , Gray Beulah H 
Published: January 2018

Abstract
Neutrophil proteases are believed to play a role in the pathogenesis of a variety of human diseases. While many studies of proteases in models of disease have focused on elastase, neutrophils contain several proteases some of which share a high degree of homology. This report describes the production and characterization of an lgG1 murine monoclonal antibody (AHN-10) that reacts with human neutrophil elastase but not with the other major neutrophil neutral proteases: cathepsin G, proteinase 3, collagenase, or the newly purified neutral protease, esterase N. AHN-10 inhibited the elastinolytic activity of purified human neutrophil elastase and could detect elastase in alcohol-fixed cytospin preparations. The epitope recognized by AHN-10 was resistant to treatment with NaIO4, suggesting that the epitope is not a carbohydrate. AHN-10 should be useful for the immunolocalization of neutrophil elastase in tissue specimens and as a stable source of characterized antibody for quantitative identification of neutrophil elastase.