bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of Colloidal Silica (Sepracell-MN) for Enrichment of Dendritic Cells From Human Peripheral Blood: Comparison With Other Methods
Authors: Chehimi Jihed , Starr Stuart E , Kawashima Hisashi , Hassan Nassef F , Douglas Steven D , Bandyopadhyay Santu 
Published: January 2018

Abstract
Three methods are described for enrichment of dendritic cells from human peripheral blood. In method 1, mononuclear cells were incubated in plastic tissue culture flasks for two h. Nonadherent cells were removed. Adherent cells were washed to remove floating cells and incubated for 14 h at 37°C in 5% CO2. Carbonyl iron was added, and the flasks were incubated for another 2 h. Nonadherent cells were subjected to centrifugation over metrizamide gradient Phagocytic cells containing ingested carbonyl iron, small lymphocytes, and free carbonyl iron particles passed through the metrizamide, while the interface cell population was enriched for dendritic cells. The purity and yield of enriched dendritic cells were 52.8% and 0.05%, respectively. In method 2, adherent mononuclear cells were cultured overnight, and the released cells (released adherent cells) were centrifuged over metrizamide to separate low-density cells. Monocytes from this low-density cell population were removed by panning over human gamma globulin-coated plastic Petri dishes. In this method the average purity and yield of DC were 63.8% and 0.1%, respectively. In method 3, released adherent cells were treated with anti-CD5 and anti-CD14 monoclonal antibodies plus baby rabbit complement for 15 min, washed, and centrifuged with colloidal silica (Sepracell-MN). Centrifugation with Sepracell-MN was repeated three times. Low-density cells were panned twice over human gamma globulin-coated plastic Petri dishes. Nonadherent cells were highly enriched for DC. Contamination of T cells, B cells, and NK cells was undetectable by flow cytofluorometry. Contamination of monocytes was &lt; 2%. This method provided &gt; 85.0% purity and 0.4% yield. This method (method 3) combines adherence, complement-dependent lysis, centrifugation with colloidal silica, and panning and provides the best yield and purity; it is therefore recommended for optimal purification of DC.
Recruitment of 99m-Technetium- or 111-Indium-Labelled Polymorphonuclear Leucocytes in Experimentally Induced Pyogranulomas in Lambs
Authors: Guilloteau Laurence , Pépin Michel , Pardon Pierre , Le Pape Alain 
Published: January 2018

Abstract
The recruitment of polymorphonuclear leucocytes (PMNs) during the development of experimental pyogranulomas induced by Corynebacterium pseudotuberculosis was followed in nine male lambs by scintigraphic examination. Autologous blood PMNs were labelled with 99m-technetium or 111-indium and were re-injected intravenously into infected lambs. The functional properties of the labelled cells were monitored 1) in vitro by measuring their phagocytic and bactericidal activity against C. pseudotuberculosis and their chemotaxis under agarose, and 2) in vivo by following scintigraphically their capacity to accumulate in an inflammatory focus induced by intradermal injection of latex beads coated with Salmonella abortus equi lipopolysaccharide. Following inoculation of corynebacteria into the right ear of lambs, radioactive foci were observed to be localized in the right ear and in the draining lymph nodes during the 4 days following inoculation. Histopathological examination performed 32 h after inoculation confirmed the intense accumulation of PMNs at these sites. With the exception of one animal, which presented visible foci in the neck 14 days postinoculation, no radioactive foci were observed during the later phases of experimental infection, despite the presence of multiple pyogranulomas which were confirmed by bacteriological examination after necropsy of the lambs. Histopathological examination of these lesions revealed layers of fibroblasts, lymphocytes, and macrophages surrounding a necrotic centre. The results of these studies suggest that the contribution of PMNs during the chronic phase of inflammation is considerably reduced in comparison with the acute inflammatory phase of the infectious process.
Rat and Human Natural Killers Exhibit Contrasting Immunoglobulin G Subclass Specificities in Antibody-Dependent Cellular Cytotoxicity Reflecting Differences in Their Fc Receptors (FcγR)
Authors: Song Elizabeth S , Young Katrina , Sears Duane W 
Published: January 2018

Abstract
Rat and human natural killers (rtNK and huNK, respectively) were compared in quantitative antibody-dependent cellular cytotoxicity (ADCC) assays for their capacity to recognize mouse and rat IgG monoclonal antibodies (MAb) of different subclasses. NK from these two species exhibit considerably different patterns of IgG subclass recognition as determined by the relative antibody concentrations required for comparable levels of target cells lysis. ADCC assays with a panel of 16 MAb revealed that the efficiency of rtNK-mediated target lysis diminished according to IgG subclass in the following order: molgG1 &gt; rtlgG2a &gt; molgG2b ≍ molgG2a &gt; rtlgG2b &gt; molgG3. By comparison, huNK recognized the same antibodies with nearly the opposite order of efficiency: rtlgG2b &gt;&gt; molgG2a &gt; molgG3 &gt; molgG2b » rtlgG2a ≍ molgG1. Only molgG2a antibodies were equally potent with rtNK and huNK. The contrasting difference in IgG subclass recognition by rat and human NK reflects the comparatively low protein sequence homology between their respective IgG Fc receptors (FcγR).
Killing of Leishmania Parasites in Activated Murine Macrophages is Based on an L-Arginine-Dependent Process That Produces Nitrogen Derivatives
Authors: Mauel Jacques , Ransijn Adriana , Buchmüller-Rouiller Yolande 
Published: January 2018

Abstract
The experiments described in this report were aimed at determining whether L-arginine (L-arg)-derived nitrogen oxidation products (nitric oxide, nitrous acid, nitrites) are involved in the intracellular killing of Leishmania parasites by activated murine macrophages in vitro. Peritoneal or bone marrow-derived macrophages were infected with L. enriettii or L. major, then activated by exposure to recombinant murine interferon-gamma or to macrophage activating factor (MAF)-rich media in the presence of li-popolysaccharide. Activation of macrophages in regular (i.e., arginine-containing) culture medium led to complete destruction of the microorganisms within 24 h (L. enriettii) or 48 h (L. major), concomitant with accumulation of nitrites (NO2-) in the culture fluids. When macrophage activation was carried out in L-arg-free medium, however, neither parasite killing nor NO2- production was obtained. A similar inhibition of macrophage leishmanicidal activity and of NO2- release was observed using media treated with arginase (which converts L-arg to urea and ornithine), or supplemented with NG-monomethyl-L-arg or guanidine (which inhibit the conversion of L-arg to nitrogen oxidation products). In all these situations, an excellent correlation between the levels of NO2- production by macrophages and intracellular killing of Leishmania was observed, whereas no strict correlation was detectable between leishmanicidal activity and superoxide production. Intracellular parasite killing by activated macrophages could be prevented by addition of iron salts to the incubation fluids. Incubation of free parasites with NaNO2 at acid pH (which permits the production of nitrous acid) led to immobilisation, multiplication arrest, and morphological degeneration of the microorganisms. Similarly, exposure of infected cells to NaNO2 led to killing of the intracellular parasite without affecting macrophage viability. These experiments strongly suggest that the leishmanicidal effect of activated murine macrophages involves the agency of L-arg-derived nitrogen oxidation products.
Effects of Interleukin 4 Upon Human Tumoricidal Cells Obtained From Patients Bearing Solid Tumors
Authors: Jadus Martin R , Good Robert W , Crumpacker Dina B , Yannelli John R 
Published: January 2018

Abstract
The use of human interleukin 4 (IL4) in the generation of tumoricidal effector cells derived from cancer patients undergoing either interleukin 2/lymphokine activated killer cells (IL2/LAK) therapy or tumor derived activated cell (TDAC) therapy was examined in the present study. Human IL4 alone did not generate LAK cells from patients undergoing IL2/LAK therapy when cultured in media containing 2% AB serum (five out of six patients). However, when the serum free medium, Aim V, was used, IL4 did generate LAK cells (eight out of nine patients), although not as cytotoxic as those activated with IL2. When cells were cultured in both IL2 and IL4 during the generation of LAK cells (3–7 days), better cell recoveries were frequently observed. The cytolytic activity of these cells against Daudi target cells was slightly reduced when compared to that response induced by IL2. This inhibition of LAK activity by IL4 was dose dependent with 1,000 U/ml IL4 producing maximal effects. This form of inhibition did not correlate with any phenotypic differences between those cells cultured in IL2 and those cells cultured in IL2 plus IL4. The IL4 mediated inhibition was also observed when the cells were cultured in Aim V medium which contains indomethacin. This inhibition induced by IL4 could not be overcome by using supra-optimal IL2. In addition to its effect during the generation of IL2 induced LAK cells, IL4 also appeared to reduce the cytolytic activity of pre-activated mature LAK cells. These results suggest that IL4 has a complex role in regulating the actions of LAK cells induced by lymphokines. When IL4 was used with IL2 during the first 5 weeks of growth of TDAC, an enhanced growth was observed when compared to the growth of TDAC when only one lymphokine was used. Besides the growth enhancement of TDAC, a better cytolytic response was observed when both lymphokines were used together. Thus, for the best growth of TDAC both IL2 and IL4 are required.
The Deficient Migration In Vitro of Lung Macrophages From Newborn Rats Normalizes After Three Days of Maturation In Situ
Authors: Kurland Geoffrey , Dauber James H 
Published: January 2018

Abstract
Because there are functional deficiencies in neonatal phagocytes, we investigated age-dependent differences in the directed migration of rat pulmonary macrophages (PM). Directed migration of PMs from newborn (&lt;24 h old) rats towards partially purified rat C5a was lower than that of PMs from adults, but reached adult levels by the third post-natal day. Calculation of the dose of C5a resulting in a half-maximal migratory response (ED-50) revealed that the ED-50s for all age groups studied were similar. These results suggest that responding PMs at all times after birth have comparable sensitivity to C5a, but that a lower percentage of newborn PMs migrate toward this factor because of defects in either signal transduction or cell motility.
Characterization of the Induction of Persistent I-A Expression by Macrophages From Bcgr Mice
Authors: Faris Mary , Zwilling Bruce S 
Published: January 2018

Abstract
Peritoneal macrophages from mice that are resistant to Mycobacterium bovis (strain BCG) [Gros et al., J. Immunol. 127,2417, 1981] can be induced to express persistently or transiently major histocompatibility complex (MHC) class II glycoproteins. The induction of persistent expression is dependent on the dose of recombinant interferon-γ (rlFN-γ). High doses of rlFN-γ (100 U) induce persistent la expression, whereas lower closes induce only transient la expression. Once induced, the persistent expression of la does not require its continued synthesis. We show that the expression of la by macrophages that transiently express MHC class II glycoproteins is reduced by the addition of cycloheximide or monensin, whereas la expression by macrophages that persistently express la is not affected. The differential sensitivity of la expression to cycloheximide was used to study the induction of persistence that is linked to the Bcg gene. Macrophages were primed with low doses of rlFN-γ in order to induce transient, cycloheximide-sensitive la expression. The cells were then treated with high doses of rlFN-γ in order to induce persistent, cycloheximide-resistant la expression. We found that the induction of persistent la expression requires at least 3 hr of exposure to rlFN-γ. Furthermore, the addition of rlFN-γ to primed macrophages is followed by a short burst of protein synthesis that is independent of the production of new mRNA. The rapidity with which persistent la expression is induced is consistent with the rapid onset of innate resistance to Mycobacterium following injection of BCG.
Biochemical and Functional Differences Between Eosinophils From Animal Species and Man
Authors: Sun F F , Crittenden N J , Czuk C I , Taylor B M , Stout B K , Johnson H G 
Published: January 2018

Abstract
Eosinophils were isolated from peritoneal lavages of repeated horse serum-injected guinea pigs or rhesus monkeys and from peripheral blood of normal human donors. Eicosanoid metabolism and chemotaxis by these cells were studied by in vitro techniques. Upon calcium ionophore stimulation guinea pig eosinophils released thromboxane B2 (TXB2) and leukotriene B4 (LTB4) while monkey and human cells produced LTC4, LTB4, and 5-HETE. Guinea pig cells do not synthesize sulfidopeptide LTs, because they lack the specific LTA4 glutathione S-transferase. Guinea pig eosinophils exhibit maximal chemotactic responses to LTB4, zymosan activated plasma, and human recombinant C5a, while producing only a negligible response to platelet activating factor (PAF). Monkey and human cells responded maximally to PAF, but exhibit only a weak response to LTB4. These results suggest that the guinea pig eosinophils differ from monkey and human eosinophils in both the synthetic capacity and functional chemotaxis responses to lipid mediators.
The Ability of Polymorphonuclear Leukocyte Priming Agents to Overcome Influenza A Virus-Induced Cell Dysfunction
Authors: Abramson Jon S , Wagner M Paige , Ralston Elizabeth P , Wei Yu , Wheeler J Gary 
Published: January 2018

Abstract
The major mortality and morbidity resulting from influenza virus infections are due to secondary bacterial infections which occur in association with virus-induced inhibition of polymorphonuclear leukocyte (PMNL) function. The present study was undertaken to determine if compounds which prime PMNL function to subsequent stimulation with N-formylmethionyl-leucylphenylalanine (FMLP) or phorbol 12-myristate 13-acetate (PMA) can overcome influenza A virus (IAV)-induced inhibition of the PMNL chemiluminescence response to these stimuli. Granulocyte-macrophage colony stimulating factor (GM-CSF), guanosine triphosphate (GTP), and 1-oleoyl-2-acetylglycerol (OAG) were able to prime the PMNL response to FMLP and/or PMA and totally or partially overcome lAV-induced PMNL dysfunction in cells stimulated with FMLP or PMA. A direct correlation was found between the extent of PMNL priming due to GM-CSF, GTP, and OAG and the capacity of these compounds to overcome virus-induced PMNL dysfunction. The implications of these findings in regard to the mechanism by which priming agents overcome lAV-induced cell dysfunction and the potential of these compounds as therapeutic agents to treat secondary bacterial infections are discussed.
Kinetics of Phagocytosis and Phagosome-Lysosome Fusion in Hamster Lung and Peritoneal Macrophages
Authors: Bizal Colette L , Butler James P , Feldman Henry A , Valberg Peter A 
Published: January 2018

Abstract
The time course of phagocytosis and phagosome-lysosome fusion (PLF) in lung and peritoneal macrophages (LMs and PMs) was measured. Lysosomes in unelicited hamster LMs and PMs were labeled with lucifer yellow. Macrophages then phagocytized heat-killed Saccharomyces cerevisiae (yeast) and were evaluated at several time points for the degree to which yeast particles were adherent vs. internalized and for the presence or absence of PLF as based on the presence or absence of lucifer yellow in yeast-containing phagosomes. A three-compartment model (adherent, ingested, fused) of independent phagocytosis and PLF was developed; the number of yeast particles in each compartment was counted, and rate constants for ingestion and fusion were determined. Comparison of rate constants showed that ingestion was significantly faster in PMs (0.047 ± 0.005 min-1) than in LMs (0.016 ± 0.005 min-1) (mean ± pooled SEM; P &lt; 0.001). Similarly, PLF was significantly faster in PMs (0.109 ± 0.013 min-1) than in LMs (0.046 ± 0.013 min-1) (P &lt; 0.003).
Colony-Stimulating Factors Increase Resistance to Atypical Mycobacteria in Resistant Mice, Whereas They Decrease Resistance in Susceptible Strains of Mice
Authors: Denis Michel 
Published: January 2018

Abstract
Inbred strains of mice, notably the susceptible C57BL/6 and the resistant A/J strains of mice, were infected with a strain of Mycobacterium avium. The infection in the visceral organs of mice was then studied, and the effect of colony-stimulating factors, i.e., interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), and macrophage colony-stimulating factor (CSF-1) on the infectious process was evaluated. Infusion of GM-CSF, CSF-1, and IL-3 led to a significant, albeit rather modest, increase in the mycobacterial resistance of A/J mice, as seen by a decrease in the number of colony-forming units (CFU) in the organs. Conversely, these CSFs dramatically increased the susceptibility of C57BL/6 mice, as seen by increased bacterial numbers in the spleens and livers. In vitro studies demonstrated that resident peritoneal macrophages from susceptible mice were more permissive than cells from resistant mice for mycobacterial growth. Application of CSFs on peritoneal macrophage monolayers led to an increased growth in both A/J and C57BL/6 monolayers for IL-3 and CSF-1 and a small microbiostatic effect for GM-CSF. Cytokine treatment did not, however, change the resistance/ susceptibility phenotype of isolated macrophages. Our results indicate that CSFs may exert beneficial or detrimental effects on resistance to mycobacteria depending on the host genetic make up.
Suppression of Human Macrophage Function In Vitro by δ9-Tetrahydrocannabinol
Authors: Specter Steven , Lancz Gerald , Goodfellow Darlene 
Published: January 2018

Abstract
The ability of macrophages to function in the presence of δ9-tetrahydrocannabinol (THC), the major psychoactive component in marijuana, was evaluated. THC added to macrophage cultures prepared from human peripheral blood inhibited macrophage spreading and phagocytosis of yeast. The effects of THC were concentration dependent, with inhibitory effects observed from 10 to 1 μg ml or lower. These results suggest that macrophages are more sensitive to THC than are lymphocytes because macrophage functions were inhibited by THC at concentrations that did not affect lymphocyte function. Thus, inhibition of lymphocyte function(s) by THC could be attributed to a direct effect of the drug on macrophages which indirectly results in lowered lymphoid cell activity.
Production of Granulocyte-Macrophage Colony-Stimulating Factor or GM-CSF-Like Growth Factor by Rat Thymic Epithelial Cell Line
Authors: Nanno Masanobu , Takahashi Tetsu , Sakata Tsuneaki , Horikawa Tatsuya , Sako Tomoyuki , Suzuki Ryuji , Itoh Tsunetoshi , Kumagai Katsuo 
Published: January 2018

Abstract
An epithelial cell line, IT-45R1, has been established from a thymus of normal Wistar rat and was found to produce growth factor which stimulated the proliferation of bone marrow cells. This growth factor induced the formation of colonies composed of macrophages, granulocytes, or both, in semi-solid medium and stimulated the proliferation of an interleukin 3 (IL3)/granulocyte-macrophage colony-stimulating factor (GM-CSF)-dependent clone. Neither IL3-dependent clone nor IL6-dependent clone responded to IT-45R1 factor. Additionally, IT-45R1 factor acted on both rat and mouse bone marrow cells but not on human bone marrow cells. Molecular weight of IT-45R1 factor was 30 kD and its isoelectric point was 4.5. To determine whether IT-45R1 factor is GM-CSF or not, Northern blot analysis and neutralization with anti-mouse GM-CSF antibody were carried out. IT-45R1 expressed GM-CSF mRNA, but neither M-CSF nor IL6 transcripts. However, antiserum specific for mouse GM-CSF did not neutralize IT-45R1 factor. Taken together, a rat thymic epithelial cell line, IT-45R1, constitutively produces GM-CSF or GM-CSF-like growth factor.
Mouse T cell clones against Mycobacterium avium: identification of clones that modify resistance against atypical mycobacteria infection
Authors: Denis Michel 
Published: January 2018

Abstract
Mouse T cell clones against live Mycobacterium avium were generated from the spleens of BALB/c mice infected with M. avium TMC 702. Eighth clones were of the L3T4+ subset, whereas two were of Lyt2+ subset. Six of the L3T4+ T cell clones were of the TH1 subset whereas two were of the TH2 subset, judged on the profile of cytokine release. One of the Lyt2+ clones exhibited significant cytotoxicity against M. avium-infected mouse macrophages. Transfer of clones to nude BALB/c mice infected with M. avium was associated with insignificant changes in resistance for seven clones. One clone, of the L3T4+/TH2 subset, transferred significant resistance to the infection, also associated with infusion of supernatants from the clone, which was fully inhibited by neutralizing with anti–interleukin 4. By contrast, infusion of one TH1 clone and the cytolytic Lyt2+ led to increased microbial growth in the spleens and livers of infected mice, which was not apparent on infusion with supernatants. Application of clones' supernatants on infected macrophages had marginal effects on M. avium growth and was not correlated with protective or suppressive activity. Overall, these results suggest that T cells may influence M. avium growth in vivo in a bidirectional manner and also suggest that interleukin 4 may be an important factor in host resistance to M. avium.
Differences in arachidonic acid metabolism by human myelomonocytic cell lines
Authors: Madden M C , Becker S , Koren H S , Friedman M 
Published: January 2018

Abstract
The production of arachidonic acid metabolites by the HL60, ML3, and U937 human phagocyte cell lines was determined after incubation with interferon-γ (IFN-γ, 500 U/ml) or vehicle for 4 days. Cells were prelabeled with tritiated arachidonic acid, [3H]AA, for 4 h, and media supernatants were analyzed by high-performance liquid chromatography. None of the cell lines produced [3H]AA metabolites in large amounts during an unstimulated, basal release period (30 or 60 min). In response to 10 μM calcium ionophore A23187 incubation (30 min), undifferentiated and IFN-γ-differentiated HL60 cells formed both cyclooxygenase products (thromboxane and prostaglandins) and lipoxygenase products (leukotrienes and hydroxyeicosatetraenoic acids). In contrast to the HL60 cells, IFN-γ-differentiated U937 cells formed primarily cyclooxygenase products and undifferentiated and IFN-γ–differentiated ML3 cells did not form any [3H]AA metabolites in response to A23187. These results indicate the need to be careful in selecting a cell line for use in a phagocyte assay system when cyclooxygenase and/or lipoxygenase products could influence the assay results.