bluuebunny/crossref_metadata_2025_split huggingface dataset:
Polyanionic Agents Inhibit Phagosome-Lysosome Fusion in Cultured Macrophages: A Reply to the Suggestion of Goren, Vatter, and Fiscus to the Contrary
Authors: D’Arcy Hart P , Young M R 
Published: January 2018

Abstract
The paper and review by Goren et al. (J. Leukocyte Biol. 41,111, 1987) contain serious objections to the reports from several laboratories on the pattern of fusion of secondary lysosomes with phagosomes (yeasts being predominantly the target) in potyanion-treated macrophages; these reports had concluded that the polyanions were inhibitors of this fusion. The main objection by Goren et al. is to the alleged misuse of electron microscopic (EM) lysosome markers; many instances of phagosome-lysosome (P-L) fusion in the treated cells have therefore been missed. The central argument is that 1) the “hydrocolloid” properties of certain of these polyanions hinder the passage of the enmeshed marker from lysosome to phagosome after their fusion and 2) this hindrance is mistakenly interpreted as indicating that fusion has not taken place, thus giving rise to the belief that the polyanions can inhibit fusion. In reply, we explain that we score as P-L fusion any instance of marker (ferritin) being seen anywhere in a fused phagosome (phagolysosome). For this crucial reason, immobilisation of marker by a hydrocolloid polyanion, e.g., in lysosomal residue of phagolysosomes or just within phagosome membranes (as in Goran’s Figs. 5 and 6 [8]), would not seriously threaten the marker distinction between fusion and nonfusion (with consequent underestimation of the former) and therefore would not invalidate the reports of a high incidence of nonfusion in potyanion-treated macrophages. Such inhibition of P-L fusion is supported by using as lysosomal label the nonpermeant fluorescent probe lucifer yellow (accepted by Goren et al. as a reliable indicator of fusion). Further support comes from the correlated inhibition of the saltatory movements of secondary lysosomes previously described; static lysosomes will have their contact with the phagosomes severely restricted. Another criticism is based on the failure of certain salient properties and functions of phagosomes to change significantly after polyanion treatment of the macrophages; these include intraphagosomal digestion, presence of lysosomal enzymes, and acidification. However, this indirect evidence can be accounted for alternatively by the operation of factors (primary lysosomes, endogenous acidification, etc.) not affected by the polyanionic block. We conclude that fusion inhibition by the polyanions is a real phenomenon, as previously reported, notwithstanding the hydrocolloid properties of some of them. Furthermore, an explanation based on hydrocolloid properties is questionable, since one or possibly two of the five main polyanionic agents appear not to be hydrocolloids.
Enrichment of Dendritic Cells From Human Peripheral Blood
Authors: Räsänen Liisa , Lehto Maili , Leinikki Pauli 
Published: January 2018

Abstract
A simplified method is described for purification of dendritic cells from human peripheral blood. The method is based on depletion of phagocytes with carbonyl iron and magnet, followed by centrifugation of nonphagocytic cells on Percoll and elimination of contaminating T lymphocytes, B lymphocytes, natural killer cells, and monocytes from the low-density cell fraction by treatment with monoclonal antibodies and complement. The purity of enriched dendritic cells was about 80% and these cells represented 0.2% of the starting mononuclear cell population. Dendritic cells were potent autologous and allogeneic stimulators in mixed leukocyte cultures.
Heterogeneity of Macrophages Evidenced by Variability in Their Glycoconjugates
Authors: Tsukada M , Spicer S S 
Published: January 2018

Abstract
Staining rat tissues with a battery of 15 lectin-horseradish peroxidase conjugates showed that macrophages contain glycoconjugates possessing terminal α, β-galactose, N-acetylgalactosamine, fucose, and N-acetylneuraminic acid, plus two terminal disaccharides. Dissimilar binding of lectins by different phagocyte populations in the same or different organs evidenced variability in glycoconjugates according to the location of the macrophages. With a group of four lectins, macrophages stained most intensely in lung, next strongest in splenic red pulp and lymph node sinuses, and weakest in skin and liver. Two populations of macrophages were newly recognized in spleen on the basis of content of fucose-rich glycoconjugate. These included a necklace-like band of macrophages at the border between marginal zone and germinal center and distinctive macrophages dispersed throughout the marginal zone. In lymph nodes, phagocytes stained strongly in the germinal centers and weakly in sinuses for glycoconjugate with N-linked oligosaccharides and conversely for glycoconjugate with terminal β-galactose. Variable lectin binding indicated heterogeneity of thymic macrophages. Lectin cytochemistry offers increased sensitivity for detecting macrophages in tissue sections, provides selective staining that shows the prevalence and distribution of the phagocytes and differentiates macrophages into separate subtypes.
Human Alveolar Macrophage Function: Differences Between Smokers and Nonsmokers
Authors: Thomassen Mary Jane , Barna Barbara P , Wiedemann Herbert P , Farmer Mark , Ahmad Muzaffar 
Published: January 2018

Abstract
Human alveolar macrophages and peripheral blood monocytes were obtained from smoking and nonsmoking normal volunteers. The macrophages and monocytes were incubated in vitro with bacterial lipopolysaccaride (LPS). The oxidative metabolic response of these cells was measured by superoxide anion production. Macrophages from smokers were suppressed in their superoxide anion response to LPS activation as compared to macrophages from nonsmokers. Monocytes from smokers and nonsmokers were not different. The cytotoxic properties of these macrophages and monocytes were assessed by an in vitro 3H-thymidine release assay against various allogeneic target cells. Macrophages and monocytes exposed to LPS were rendered tumoricidal. Macrophages from nonsmokers appeared to generate greater cytotoxic activity than macrophages from smokers. Macrophages from both smokers and nonsmokers were cytotoxic for three different tumorigenic cell lines but not for a nontumorigenic cell line. Monocytes from smokers and nonsmokers were not different in cytotoxic activity. We conclude that macrophages from both smokers and nonsmokers can be activated after exposure to LPS; however, macrophages from smokers may be slightly suppressed in their responses.
Interferon-lnduced Indoleamine 2,3-Dioxygenase Activity in Human Mononuclear Phagocytes
Authors: Carlin Joseph M , Borden Ernest C , Sondel Paul M , Byrne Gerald I 
Published: January 2018

Abstract
Interferon (IFN)-induced tryptophan degradation, catalyzed by indoleamine 2,3-dioxygenase (IDO), has been shown to mediate antimicrobial activity in epithelial cells. IDO activity has also been augmented in peripheral blood mononuclear cells (PBMC) treated with IFN or interleukin-2 (IL-2). The effector cells in this population have now been further characterized. PBMCs were isolated from normal donors, separated into monocyte and lymphocyte populations by plastic adherence, treated with IFN or IL-2, and cultivated in medium supplemented with [3H]tryptophan. Culture supernatants were collected after a 48-h incubation and fractionated by high-performance liquid chromatography; radioactivity was determined in fractions corresponding to tryptophan and its metabolites. IFN-γ and IFN-β induced IDO activity only in monocytes (plastic-adherent, nonspecific esterase-positive PBMCs). The induction of IDO activity by IL-2 required both monocytes and lymphocytes. Interaction was required between these populations for induction of IDO by IL-2, due to production of IFN-γ by T lymphocytes, with subsequent IFN-γ-mediated induction of IDO in monocytes. A number of myeloid cell lines as well as monocyte-derived macrophages were also tested for their ability to be induced to degrade tryptophan in response to IFN treatment. Monocyte-derived macrophages were found to retain their capacity to be induced by IFN-γ and IFN-β to degrade tryptophan after differentiation, and to possess seven times more IDO activity per cell than IFN-induced monocytes. However, the presence of lipopolysaccharide (LPS) in the culture medium was required for the maximum induction of IDO activity by IFN-β. Furthermore, higher concentrations of LPS were sufficient to induce IDO activity in macrophages in the absence of exogenous IFN.
Capacities of a Newly Established Thymic Stromal Cell Clone to Express la Antigens and to Produce Interleukin-6, Colony-Stimulating Factor, and Thymic Stroma-Derived T-Cell Growth Factor
Authors: Ogata Masato , Matsubara Hidetoshi , Takai Yasuyuki , Kosaka Hiroshi , Katagiri Tatsuo , Sano Haruo , Ishimura Kazunori , Fujita Hisao , Hamaoka Toshiyuki , Fujiwara Hiromi 
Published: January 2018

Abstract
Thymic stromal cell lines, termed MRL104 and MRL28, have been isolated from long-term liquid cultures of thymic stromal cells from MRL/1 mice. The capacities of these parental lines and derived clones to express la antigens and to produce cytokines involved in T-cell proliferation and/or differentiation were investigated. Parental lines and their clones did not exhibit a typical fibroblastic, macrophage-like, or epithelial appearance in electron as well as phase-contrast micrographs. These thymic stromal cells seemed to differ from established fibroblast lines in that these thymic stromal cells expressed la antigens after exposure to interferon-γ (IFN-γ), whereas fibroblast lines did not They also appeared to differ from macrophage cell lines in that they lacked the expression of Mac-1 antigens on their cell surface and produced no detectable level of interteukin-1 (IL1) before or even after exposure to lipopolysaccharide. When these parental lines and its clones were tested for their ability to produce various types of cytokines, it was revealed that they were capable of producing colony-stimulating factor (CSF), IL6, and thymic stroma-derived T cell growth factor (TSTGF). which was recently described, but were unable to generate other lymphokines and IFNs. Thus these cell lines and clones represent unique features in that they have potentials to express la antigens and to produce CSF, IL6, and TSTGF. The biological significance for the expression of these features is discussed in the context of intrathymic T-cell maturation and T-cell repertoire selection.
Oxidative Responses of Rabbit Alveolar Macrophages: Comparative Priming Activities of MIF/MAF, Sera, and Serum Components
Authors: Hayakawa Hiroshi , Umehara Keiko , Myrvik Quentin N 
Published: January 2018

Abstract
The comparative abilities of various reagents to prime rabbit alveolar macrophages (AM) to produce reactive oxygen intermediates (ROI) in a chemiluminescent (CL) assay were investigated. It was noted that AM from normal rabbits cultured in a serum-free medium for 18 hr exhibited a “spontaneous” priming response following a challenge with phorbol myristate acetate (PMA); however, “spontaneous” priming was not evident when the AM were cultured for only 3 hr. It was further established that pretreatment of normal AM for 3 or 18 hr with MIF/MAF preparations (serum-free), fetal bovine serum (FBS), or bovine serum albumin (BSA) exhibited marked increases in their CL reponses following challenge with PMA. When FBS was used in the culture medium, the priming activity of MIF/MAF was masked because of the high CL responses of controls due to the priming effects of FBS. BSA at concentrations approximately equivalent to the amount in FBS also displayed marked priming activity. Bacterial products (lipopolysaccharide and muramyl dipeptide), latex particles, rabbit IgG, PMA, and opsonized as well as nonopsonized zymosan and bacteria (BCG and Staphylococcus epidermidis) were inactive as priming agents. In comparison, AM from BCG-immune rabbits that were primed in vivo yielded a very large CL response when challenged with PMA. Opsonized zymosan and bacteria produced twofold increases in the CL responses in BCG-immune AM compared to nonopsonized preparations. The marked priming effect of serum on AM cultured for even a short period (3 hr) indicates that normal AM undergo marked changes in culture that complicate the interpretation of AM function when AM are cultured in vitro in media containing serum.
Interactions of Pseudorabies Virus With Swine Alveolar Macrophages: Effects of Virus Infection on Cell Functions
Authors: Iglesias Gerardo , Pijoan Carlos , Molitor Thomas 
Published: January 2018

Abstract
In order to assess the effect of Pseudorabies virus (PRV) infection on the function of swine alveolar macrophages (AM), lung lavage cells were cultured, infected with one of six strains of PRV, and various activities were measured. Activity measurement included viability, phagocytosis of yeast, phagosome-lysosome fusion, phagocytosis of opsonized particles, and superoxide release. AM were infected with 5 × 10-3 PFU/cell, and the comparative assessment of functions was performed at 18-20 h postinfection. Cell viability in PRV-infected cultures ranged from 79 to 94% of the viability in noninfected cultures. Phagocytosis of yeast was significantly reduced only in the AM cultures infected with the strain S-62. Phagosome-lysosome fusion was depressed in cultures infected with the strains S-62, 4892, 3816, and BUK. The phagocytosis of opsonized sheep red blood cells showed significant differences between noninfected and PRV-infected cultures in all cases except cultures infected with the strain PRV-C. The O2 release after stimulation with opsonized zymosan was significantly reduced in all the PRV-infected cultures. The effect of PRV infection on AM functions that are related to the bacterial activity of such cells suggests that PRV-induced AM dysfunction might have a role in the increased susceptibility of PRV-infected pigs to bacterial pneumonia.
Separation of Potent and Poorly Functional Human Lung Accessory Cells Based on Autofluorescence
Authors: Nicod Laurent P , Lipscomb Mary F , Toews Galen B , Weissler Jonathan C 
Published: January 2018

Abstract
Human alveolar macrophages obtained by bronchoalveolar lavage are usually poor accessory cells in in vitro lymphoprollferation assays. However, we recently described a subpopulation of pulmonary mononuclear cells, obtained from minced and enzyme-digested lung, which were potent stimulators of allogeneic T-lymphocyte proliferation. These cells were enriched in loosely adherent mononuclear cell (LAM) fractions, but further study of these accessory cells was hampered by the heterogeneous nature of LAM. It was observed that in the majority of lung tissue sections, most alveolar macrophages were autofluorescent, whereas most interstitial HLA-DR positive cells were not. Therefore autofluorescence was utilized to fractionate LAM in an attempt to remove alveolar macrophages and selectively purify interstitial accessory cells. LAM were separated by flow cytometry using forward and side scatter to exclude lymphocytes, and red autofluorescence to obtain brightly autofluorescent (A pos) and relatively nonautofluorescent (A neg) mononuclear cells. Although both populations contained over 80% HLA-DR positive cells, A pos cells were poor accessory cells, whereas A neg cells were extremely potent stimulators of a mixed leukocyte reaction at all stimulator ratios tested. When A pos cells were added to A neg cells, T-cell proliferation was markedly suppressed in the majority of experiments. Morphologically, A pos cells appeared similar to classical alveolar macrophages with 95% of the cells being large and intensely nonspecific esterase positive. In contrast, the majority of A neg were smaller, B-cell antigen-negative, nonspecific esterase negative, and had a distinctive morphology on Wright-stained smears. We conclude that fractionation of LAM based on autofluorescence is a powerful tool to isolate and characterize lung mononuclear cells that act either as stimulators or as suppressors of immune responses in the lung.
Characteristics of Monocyte Angiotensin-Converting Enzyme (ACE) Induction by Dexamethasone
Authors: Vuk-Pavlović Zvezdana , Kreofsky Teresa J , Rohrbach Michael S 
Published: January 2018

Abstract
Monocyte maturation to macrophages and transformation into epithelioid granuloma cells in some granulomatous diseases are accompanied by the induction of membrane-bound angiotensin-converting enzyme (ACE). The physiologic and pathophysiologic roles of ACE generated in these processes are not known. The pattern and the mechanism of ACE induction in human monocytes are also not well understood. Dexamethasone is one of the agents reported to induce elevated ACE activity in human monocytes, and therefore a suitable tool for studying the phenomenon. This study shows that dexamethasone augments monocyte ACE in a Diphasic dose-dependent manner with maximum effect at 10-5 M concentration. Although it enhances the level of ACE activity, dexamethasone doss not alter the time course for ACE induction from that found in unstimulated monocytes. The ACE activity of monocytes cultivated in 10 nM dexamethasone and then exposed to 10-3 M diazosulfanilic acid (DASA) is reduced approximately by 80% in comparison with cells not treated with DASA, demonstrating that dexamethasone-induced ACE is an ectoenzyme. Dexamethasone does not increase the activity of other monocyte ectoenzymes: γ-glutamyltransferase, alkaline phosphodiesterase-I, and leucine aminopeptktase, showing that dexamethasone induction of ACE is a specific, rather than generalized, effect on plasma membrane enzymes. It is suggested that the increase in ACE activity is due to the increased rate of enzyme synthesis.
Lithium Chloride Stimulates Human Monocytes to Secrete Tumor Necrosis Factor/Cachectin
Authors: Kleinerman Eugenie S , Knowles Rebecca D , Blick Mark B , Zwelling Leonard A 
Published: January 2018

Abstract
The purpose of this study was to examine the effect of lithium chloride (LiCI) on human monocytes. Patients undergoing lithium therapy have elevated white blood cell counts. Since both tumor necrosis factor alpha (TNFα) and interleukin 1 (IL-1), which are secreted by monocytes, can stimulate endothelial cells to produce granulocyte-macrophage colony-stimulating factor (GM-CSF), we determined whether lithium-stimulated monocytes produced TNFα and/or IL-1. Normal human monocytes were incubated for 24 h with medium (negative control), lipopolysaccharide (positive control), or LiCI (0.05–50 mM). The supernatants were removed and assayed for IL-1 and TNFα secretion using the D10.G4.01 and L929 assays, respectively. Lithium did not stimulate IL-1 secretion but did stimulate TNFα secretion (5–10 U/ml of TNFα per 2 x 105 monocytes). The increased secretion of TNFα was associated with a fourfold increase in TNFα mRNA. TNFα activity in the supernatants was neutralized by a monoclonal antibody against human TNFα but not by antibody against human albumin. Other alkali metals such as rubidium and cesium did not stimulate monocytes to secrete TNFα. These data indicate that one mechanism by which Li may cause granulocytosis is through a transcriptional enhancement of TNF production and subsequent secretion by monocytes.
Role of C5a in the Induction of Tumoricidal Activity in C3H/HeJ (Lps
d) and C3H/OuJ (Lps
n) Macrophages
Authors: Hogan M Michele , Yancey Kim B , Vogel Stefanie N 
Published: January 2018

Abstract
Thioglycollate-elicited macrophages from C3H/HeJ (Lpsd) and C3H/OuJ (Lpsn) mice were cultured in a two-signal, tumoricidal assay using recombinant interferon-γ (rlFN-γ) as the “priming” signal and recombinant human C5a (rC5a) as the “trigger” signal. These experiments were compared directly with a well established, two-signal tumoricidal assay in which rlFN-γ was used as the “priming” signal and protein-rich, butanol-extracted lipopolysaccharide (But-LPS) as the “trigger” signal. These studies showed that rlFN-γ-primed macrophages can be triggered in a dose-dependent manner by rC5a to effecthigh levels of tumoricidal activity. Maximum levels of cytotoxicity achieved using this endogenously produced, biologically active peptide as a “trigger” signal were comparable to those obtained using But-LPS. Moreover, experiments in which anti-C5 antibody was included in macrophage cultures stimulated with rlFN-γ and But-LPS showed a significant reduction (P &lt; .05) in tumoricidal activity. Because LPS has been shown to induce macrophage C5 production and enzyme release, these findings suggest that macrophage-derived C5 is locally converted to C5a (or some other biologically active C5 cleavage fragment), which functions as an autocrine trigger signal for the induction of tumoricidal activity. In summary, these data suggest 1) that rC5a can provide a “second signal” to rlFN-γ-primed murine macrophages for the induction of tumoricidal activity and 2) that macrophage-derived C5 or C5a may represent an autocrine signal induced by exogenous “trigger signals.”
Natural Killer Cell Activity and Macrophage-Dependent Inhibition of Growth or Killing of Mycobacterium avium Complex in a Mouse Model
Authors: Bermudez Luiz Eduardo M , Kolonoski Peter , Young Lowell S 
Published: January 2018

Abstract
Natural killer (NK) cells from spleens of normal and Mycobacterium avium complex (MAC)-infected C57 black mice (C57 BL/6 bg/+) were examined for their capacity to activate splenic and peritoneal macrophages from beige mice to inhibit or kill intracellular MAC. Peritoneal and splenic macrophages from beige mice were exposed in vitro to NK cells obtained from MAC-infected and uninfected black mice. NK cells from uninfected black mice were also treated in vitro with recombinant interleukin-2 (IL-2) for 48 h before incubation with macrophages. While control macrophages supported intracellular growth of MAC, macrophages exposed to unactivated NK cells inhibited growth of the intracellular bacteria, as determined 4 days after infection. IL-2 stimulated NK cells, and NK cells obtained from MAC-infected animals were able to activate murine macrophages in vitro to inhibit growth or kill 40.0 ± 5% and 61.3 ± 6% of the intracellular bacteria, respectively. In other experiments, beige mice (C57 BL/6 bg/bg) were treated intraperito-neally with NK cells obtained from MAC-infected and uninfected C57 black mice. Peritoneal macrophages harvested from beige mice treated with NK cells activated in vitro with IL-2 killed 24.4 ± 4% of intracellular bacteria by day 4 after infection. Macrophages obtained from animals treated with NK cells harvested from MAC-infected black mice killed 58.8 ± 7% of intracellular bacteria by 4 days after infection, in contrast with intracellular growth observed in macrophages obtained from untreated animals and from animals treated with Hanks' solution or unactivated NK cells. These crossover studies suggest that NK cells may be important in host defense against MAC.