bluuebunny/crossref_metadata_2025_split huggingface dataset:
Volume Dependent Polymorphonuclear Leukocytes Fractions Isolated by Counterflow Centrifugal Elutriation: PMN Volume Does Not Correlate With PMN Age
Authors: Berkow Roger L , Dodson Robert W 
Published: January 2018

Abstract
We have previously reported that human peripheral blood polymorphonuclear leukocytes can be separated into at least six volume dependent fractions using counterflow centrifugal elutriation (CCE). Larger volume was shown to correlate with increased superoxide release with either the chemotactic peptide F-met-leu-phe (FMLP) or the tumor promotor phorbol myristate acetate (PMA). To help explain these findings we have examined volume dependent PMN fractions (VDPF) for PMN functions felt to be correlated to PMN age, ie, alkaline phosphatase activity, phagocytosis of opsonized particles, adherence to plastic, and directed movement to FMLP and zymosan activated serum. PMN alkaline phosphatase activity was found to be low in the smallest PMNs, increasing in the PMNs of intermediate size, and then decreasing again in the largest PMN functions. A similar relationship was noted for PMN phagocytic activity. No significant difference among VDPF in adherence to plastic or Chemotaxis was found. Furthermore, no difference among VDPF was seen for receptor number or binding affinity of the ligands FMLP or phorbol dibutyrate. Total cellular activity of the NADPH dependent oxidase was, however, 200% greater in the largest compared to the smallest VDPF. Because of the lack of consistent correlation between “age” related PMN function and PMN volume, it is unlikely that PMN volume represents a manifestation of PMN age. It is likely that the increased oxidase activity seen among larger VDPF accounts for the more rapid oxidative burst previously noted.
Cells of the Human Myelomonocytic Line RC-2A Synthesize Tissue Factor-Like Procoagulant and Urokinase-Type Plasminogen Activator
Authors: Karhumäki E , Saravo L , Helin H 
Published: January 2018

Abstract
Cells of the myelomonocytic leukemia cell line RC-2A were studied for their ability to synthesize clotting-promoting and fibrinolytic factors. The cells were observed to generate procoagulant activity (PCA) in readily measurable quantities. Incubation of RC-2A cells with phorbol myristate acetate (PMA; 3 ng/ml) or Phytohamagglutinin (PHA, 10 μg/ ml) for 18 h resulted in a 4-5-fold increase in PCA relative to unstimulated control. The PCA of RC-2A cells was tissue factor-like in that it was dependent on factor VII but not on factors VIII or IX. RC-2A cells also produced plasminogen activator (PA). Secreted PA was approximately 70% of the PA of an identical number of human monocyte-derived macrophages; fresh isolated monocytes synthesized virtually no PA. Compared to macrophages, RC-2A cells secreted less or no PA-inhibitors. Lysates of RC-2A cells contained over three times more PA than lysed macrophages. Stimulation of the cells with lectins (PHA, concanavalin A) or PMA was followed by a modest (2-3-fold) increase in PA. Enzyme immunoassay with antibodies to urokinase (u-PA) or tissue-type PA (t-PA) identified the RC-2A plasminogen activator as being of urokinase type.
Effects of Acute Graft-vs-Host Disease on the Liver of the Brown Norway Rat
Authors: Hightower James A , Earnest David L , Martens Anton C M , Zurcher Chris , Brouwer Adriaan , Blauw Elizabeth , de Leeuw A Margreet , Hagenbeek Anton 
Published: January 2018

Abstract
In this study we examined the effects of acute graft-vs-host disease (aGVHD) on the Brown Norway (BN) rat liver. When clinical signs of the disease appeared, rats were inoculated with fluorescent latex beads and 30 min later nonparenchymal cells were isolated from the liver. The cells were then analyzed via flow cytometry, histochemistry, and electron microscopy. Flow cytometry demonstrated that 58% of the cells from the 80 ml/min elutriation fraction (normally rich in Kupffer cells) of the non-GVHD liver had high fluorescence intensity compared to 8% in rats with aGVHD. Determination of the cellular composition of the various fractions with electron microscopy confirmed flow cytometry observations in that only 9% of the 80 ml/min elutriation fraction of GVHD livers had peroxidase-positive rough ER and the morphological appearance of macrophages as compared to 60% in the non-GVHD liver. The low percentage of fluorescent-positive Kupffer cells in the 80 ml/min elutriation fraction of the GVHD liver is attributed to a massive lymphocytic invasion of the liver and not necessarily to a defect in the mononuclear phagocyte system.
Macrophage Migration Inhibition Induced by MDP, LPS, PMA, and MIF/MAF: Reversal by Macrophage Migration Enhancement Factor (MEF), L-Fucose, L-Fucosyl BSA, D-Mannose, and D-Mannosyl BSA
Authors: Gordon Mark R , Chida Kingo , Takata Isao , Myrvik Quentin N 
Published: January 2018

Abstract
Our data establish that migration inhibition factor (MIF) and migration enhancement factor (MEF) mutually neutralize the effect of each other in a concentration-dependent manner. The monosaccharides L-fucose and D-mannose were also shown to reverse MIF and additionally to stimulate alveolar macrophage (AM) migration in the absence of MIF. The specific activity of these sugars was increased 200-fold when conjugated to bovine serum albumin (BSA). Macrophage activation is usually observed concurrently with migration inhibition when macrophages are incubated with MIF preparations. Migration inhibition occurred also when AM were incubated in the presence of known metabolic activators (MDP, PMA, and LPS). It was found that L-fucose, D-mannose, L-fucosyl BSA, and D-mannosyl BSA could reverse migration inhibition caused by MIF as well as by these metabolic activators. These observations suggest that reversal of MIF by L-fucose is unexplained solely on the basis that L-fucose is functioning as a competitive inhibitor; instead, they suggest that MEF and the above sugars and their conjugates stimulate AM through a receptor system different from the MIF receptor. These observations support the concept that MEF is an important macrophage modulator in CMI responses.
Role of Colony-Stimulating Activity in Antitumor Activity of Lactobacillus casei in Mice
Authors: Shimizu Takako , Nomoto Koji , Yokokura Teruo , Mutai Masahiko 
Published: January 2018

Abstract
Wide-ranging differences were observed between the antitumor activities of 23 lactoba-cilli (13 species; 23 strains) and their capacities to elevate the level of serum colony-stimulating activity (CSA) by intraperitoneal administration in mice, and a good correlation existed between the two activities. The mechanism of enhanced production of CSA by administration of Lactobacillus casei YIT 9018 (LC 9018), one of the bacteria that had the strongest activities, and the role of CSA in antitumor activity of LC 9018 were studied. Colony-stimulating activity in the washing fluid from the peritoneal cavity of mice that had been administered LC 9018 intraperitoneally was elevated at 3 to 24 h after the injection, and CSA was also detected at elevated levels in the serum of the mice 6 to 12 h after injection. The cells responsible for the production of CSA after stimulation with LC 9018 seem to be the resident macrophages at the site of administration, because the resident macrophages of mice lavaged 1 h after an intraperitoneal administration of LC 9018 released CSA when they were cultured in vitro. Moreover, resident peritoneal macrophages of normal mice cultured with LC 9018 in vitro also produced CSA. Similar results were obtained with athymic nude mice, and the CSA-inducing activity of LC 9018 was diminished in the mice pretreated with carrageenan, which is selectively toxic to mature macrophages. Bone marrow cells matured to macrophages and polymorphonuclear cells by culture with the CSA induced by LC 9018 for 7 days. These matured macrophages showed strong antitumor activity both in vivo and in vitro. These results suggest that CSA plays important roles in the antitumor activity of LC 9018: it enhances not only the multiplication of committed precursor cells for macrophages and polymorphonuclear cells, but also the functional maturation of the precursor cells for macrophages which serve as potent effectors for tumor cells.
Protein Phenotypes of Mouse Macrophages Activated In Vivo for Tumor Cell Killing
Authors: MacKay Robert J , Russell Stephen W 
Published: January 2018

Abstract
We have shown previously that the cellular proteins 47b and 71/73 could be used to construct phenotypes that distinguish among bone marrow culture-derived (BMCD) macrophages that were either unstimulated or primed by gamma interferon or fully activated for tumor cell killing by gamma interferon in combination with lipopolysaccha-ride (LPS). In the present study we examined in vivo-derived correlates for each of these stages of macrophage activation and found that the same protein phenotypes held true: both p47b and p71/73 were expressed by cytolytic peritoneal macrophages, including macrophages from a tumoral effusion, whereas macrophages primed in vivo by the intraperitoneal injection of either concanavalin A or methyl vinyl ether copolymer II expressed p47b but lacked p71/73. Neither resident nor inflammatory macrophage populations expressed p47b, and acute inflammatory macrophages, like unstimulated BMCD macrophages, expressed little or no p71/73. By contrast, resident and thioglycollate-elicited macrophages synthesized moderate levels of p71/73. When p71/73 also was expressed, there was a quantitative relationship between p47b expression and cytolytic activity in five different in vivo-activated macrophage populations. The results suggest that, regardless of the macrophage source or stimulus, it may be possible to assess macrophage activation status by reference to protein phenotypes utilizing p47b and p71/73.
Nisoldepine Inhibits Formyl-Methionyl-Leucyl-Phenylalanine (fMet-Leu-Phe) Receptor-Coupled Calcium Transport in Human Neutrophils
Authors: Williamson Kim C , Tauber Alfred I , Navarro Javier 
Published: January 2018

Abstract
The formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe) dependent Ca2+ uptake by human neutrophils consists of at least two components, one of which is sensitive to dihydropyridine derivatives. Inhibition by dihydropyridine derivatives showed the rank order of nisoldepine &gt; nitrendipine &gt; nimodepine &gt; /Bay K 8644. The nisoldepine-sensitive calcium uptake exhibited an ID50 of 1.5 μM and maximal inhibition were observed at 5 μM. Neither calcium efflux or [3H]fMet-Leu-Phe binding was affected by nisoldepine up to 10 μM. The inhibition by nisoldepine was inversely proportional to the extracellular calcium concentration. Unlabeled nisoldepine and other dihydropyridine derivatives displaced the specific binding of [3H]PN 200-110 to human neutrophils. Our data suggest a relationship between dihydropyridine binding and the inhibition of fMet-Leu-Phe-dependent Ca2+ uptake.
Cyclophosphamide-Induced Alterations in Human Monocyte Functions
Authors: McBride W H , Hoon D B , Jung T , Naungayan J , Nizze A , Morton D L 
Published: January 2018

Abstract
This investigation was designed to study the effects of relatively low doses of cyclophosphamide (CY) on monocyte function in patients with surgically resected melanoma. Monocytes taken from patients 3 days after receiving 300 mg, 150 mg, or 75 mg CY/m2 had decreased interleukin-1 (IL-1) production. Production of tumor necrosis factor (TNF)-like molecules by the same monocytes appeared to be enhanced following 300 mg/m2 CY but not after 150 or 75 mg/m2 CY. In vitro studies of the direct effects of CY metabolites (mafosfamide and 4-hydroperoxycyciophosphamide) on human monocytes showed only concomitant decreases in production of IL-1 and TNF-like molecules. This occurred at concentrations that did not obviously affect viability, although monocyte spreading was inhibited. No evidence was obtained for in vitro enhancement of TNF-production. We conclude that CY can affect monocyte function. In vivo it may have both direct effects leading to decreased TNF and IL-1 production and indirect effects through lymphocytic or haematopoietic systems that activate monocytes to enhanced TNF production. The effects are dose-dependent. These CY-induced changes could be responsible in part for some of the alterations in host immunity and tumor resistance that follows administration of the drug.
Tumor Necrosis Factor-α Induces Increased Hydrogen Peroxide Production and Fc Receptor Expression, but Not Increased la Antigen Expression by Peritoneal Macrophages
Authors: Hoffman Maureane , Weinberg J Brice 
Published: January 2018

Abstract
It has recently been shown that tumor necrosis factor (TNF) induces increased la antigen expression on a malignant murine macrophage cell line, and that TNF is synergistic with gamma interferon (IFN) in inducing la expression. This finding raises the possibility that TNF serves as a non-interferon macrophage activating factor in vivo. Since it is known that TNF has different effects on malignant and benign cells, we chose to evaluate the effects of recombinant TNF on primary cultures of murine peritoneal macrophages (MP). Neither human nor murine TNF increased the proportion of MP which expressed la antigen, and TNF actually partially prevented the IFN-induced increase in la. However, culture with TNF activated MP for increased hydrogen peroxide production in response to phorbol myristate acetate (PMA) and antagonized the IFN-induced decrease in the proportion of the MP bearing receptors for the Fc region of IgG2b. TNF and IFN were additive in increasing peroxide production. Both human and murine TNF had the same effects on MP, and MP from C3H/HeN (endotoxin sensitive) and C3H/HeJ (endotoxin resistant) mice responded comparably to TNF and IFN. Our results support the hypothesis that TNF has some macrophage-activating activity, but its effects are selective and not identical with those of IFN.
Dissociation of the 47-Kilodalton Protein Phosphorylation From Degranulation and Superoxide Production in Neutrophils
Authors: Sha’afi R I , Molski T F P , Gomez-Cambronero J , Huang C -K 
Published: January 2018

Abstract
The aim for the present studies is to examine the relationship between the phosphorylation of the 47-kDa protein and some neutrophil responses such as degranulation, the synergistic effect of PMA on calcium ionophore-induced degranulation, superoxide generation, and the priming of the oxidative burst produced by the chemotactic factor fMet-Leu-Phe and phorbol 12-myristate 13-acetate (PMA).
Incubation of neutrophils with the protein kinase inhibitor 1-(5-isoquinoline-sulfonyl)-2-methylplperazine (H-7) inhibits the phosphorylation of the 47-kDa protein produced by PMA and fMet-Leu-Phe but does not affect lysozyme release induced by the same stimuli or fMet-Leu-Phe-induced N-acetyl-β-glucosaminidase release. Furthermore, the synergistic effect of PMA on the A23187-induced degranulation is not inhibited by H-7. Also, pretreatment of the cells with H-7 inhibits superoxide production produced by PMA but not by fMet-Leu-Phe. The inhibitory effect of H-7 is more pronounced on the rate than on the extent of PMA-induced superoxide release. On the other hand, N-(2-guanidinoethyl)-5-isoquinolinesulfonamide hydrochloride, HA-1004, a less potent protein kinase inhibitor, has no Inhibitory effect on superoxide generation produced by fMet-Leu-Phe or PMA. In the case of superoxide production, the addition of low concentrations of PMA to rabbit neutrophils primes these cells to the subsequent stimulation by fMet-Leu-Phe, dramatically increasing the effect. Conversely, low concentrations of fMet-Leu-Phe prime the cells to PMA stimulation. The stimulation by PMA of cells primed with fMet-Leu-Phe is inhibited by H-7. Moreover, the priming effect by PMA is also inhibited by H-7. This inhibition is less pronounced at a low concentration of PMA. On the other hand, in the case of human neutrophils, the priming effect of PMA is not inhibited by H-7. These results suggest several points. First, phosphorylation of the protein identified on two-dimensional gel electrophoresis as having a molecular weight of 47 kDa and PI of 4.9 is not necessary for degranulation produced by either PMA or fMet-Leu-Phe. Second, the phosphorylation of the 47-kDa protein is not necessary for superoxide generation, at least in the case of fMet-Leu-Phe and possibly for other stimuli. Third, in rabbit neutrophils, both the priming and stimulation of superoxide production with PMA are inhibited by H-7. In human neutrophils, the priming by PMA is not affected by H-7. In the case of human neutrophils, it is reasonable to conclude that the priming of the oxidative burst by PMA does not appear to be mediated by the phosphorylation of the 47-kDa protein.
Hepatocyte Injury During Post-Operative Sepsis: Activated Neutrophils as Potential Mediators
Authors: Holman John M , Saba Thomas M 
Published: January 2018

Abstract
We evaluated the concept that the vascular entrance of both bacterial and nonbacterial participate material could lead to hepatic parenchyma) cell injury, either due to post-phagocytic Kupffer cell activity or the margination of activated leukocytes in the liver. Injection of denatured, collagen-coated particles as well as heat-killed bacteria were used as particulate challenges. Hepatic parenchymal cell injury in vivo during postoperative sepsis was evaluated by plasma aspartate aminotransferase (AST) and ornithine carbamyl transferase (OCT) enzyme levels over 3-72 h. AST and OCT levels elevated following either laparotomy plus cecal ligation (mild sepsis) or laparotomy plus cecal ligation with puncture (severe sepsis), with the peak level at 24 h. In addition, the direct intravenous injection of either nonbacterial foreign particles or heat-killed Pseudomonas aeruginosa into normal rats also produced a dose-dependent elevation of AST and OCT. The plasma level of either AST or OCT actually increased 350-400% after injection of the non-bacterial particles. A similar dose related elevation in enzymes followed the intravenous injection of heat-killed Pseudomonas. To differentiate the potential contribution of activated hepatic Kupffer celts versus activated marginated neutrophils to the in vivo hepatic injury, we determined the release of the hepatic specific enzyme OCT by cultured hepatic parenchymal cells when they were exposed to isolated Kupffer cells or isolated PMNs that were activated by exposure to dead bacteria. Bacteria alone when added to cultured hepatocytes did not induce significant OCT release. In contrast, activated PMNs but not Kupffer cells induced a significant (p &lt; 0.05) release of OCT from parenchymal cells into the culture media. Thus, in vivo transient hepatic parenchymal cell injury with post-operative sepsis may be mediated by the margination of activated PMNs in the liver.
Characterization of the Natural Suppressor Cell Population in Adult Rat Bone Marrow
Authors: Noga Stephen J , Wagner John E , Horwitz Louis R , Donnenberg Albert D , Santos George W , Hess Allan D 
Published: January 2018

Abstract
Natural suppressor cell activity (NSCA) has been ascribed to a subset of cells present in human and murine hematopoietic tissues which can suppress a variety of lymphocyte responses without MHC restriction. We investigated NSCA in lymphocyte-depleted rat bone marrow (BM) which is used as a model for prevention of graft vs host disease (GVHD) following allogeneic BM transplantation (BMT). The T-cell depleted fraction obtained after elutriation contained higher levels of NSCA than the unseparated BM. Further separation of this graft fraction by discontinuous Percoll gradient centrifugation revealed high levels of radiosensitive NSCA in the low density (&lt; 1.070) fraction which represented 0.5% of the original BM population. These cells were of blast morphology, stained intensely with a dansylated derivative of cyclosporine A (dans CsA) and weakly expressed macrophage/granulocyte antigens and non-specific esterase (NSE). These cells were initially non-adherent but proliferated in culture to produce intensely NSE positive, adherent, phagocytic cells of macrophage morphology. We conclude that the highly suppresses, radiosensitive cell present in rat BM may be of early progenitor or monocyte lineage. The grafting of natural suppressor (NS) cells and progenitor cells may affect graft/host immunoregulation and their characterization may provide insight into GVH biology and graft rejection.
Effect of Lead on Macrophage Function
Authors: Kowolenko Michael , Tracy Leigh , Mudzinski Stanley , Lawrence David A 
Published: January 2018

Abstract
Lead (Pb) has been shown to alter various parameters of immune function such as host resistance and antibody formation. In addition, various heavy metals have been implicated as inducers of autoimmunity. In these experiments, macrophages, isolated from the peritoneal cavity of mice exposed to various doses of lead in vivo as well as cells exposed in vitro were tested for the following immunologic parameters: phagocytosis, antigen presentation, interleukin 1 production, and their ability to stimulate the autologous mixed lymphocyte reaction (AMLR). The results obtained indicate that Pb appears to alter the ability of macrophages to present antigen by enhancing the AMLR while having no effect on phagocytosis or IL-1 production. These data suggest that Pb may interfere with antigen-specific interactions between macrophages and T cells.
An Abnormal Calcium Uptake Pump in Chediak-Higashi Neutrophil Lysosomes
Authors: Styrt Barbara , Pollack Charles R , Klempner Mark S 
Published: January 2018

Abstract
Calcium is mobilized from intracellular stores during phagocyte activation and appears to be involved in stimulus-response coupling in these and other cell types. Because the lysosome is a calcium-sequestering organelle in the neutrophil and abnormal lysosome morphology is associated with defective neutrophil function in Chediak-Higashi syndrome (CHS), we examined ATP-dependent calcium uptake in neutrophil lysosomes from the beige mouse model of CHS. We present findings indicating that CHS lysosomes have an enhanced capacity for ATP-dependent calcium uptake relative to control lysosomes. Kinetic analysis showed differences in Vmax and in the Km for both ATP and calcium, suggesting that both the number of lysosomal calcium uptake pumps and their substrate affinity may be altered in CHS. We conclude that a genetically determined abnormality of a subcellular calcium transport system may contribute to the structural and functional defects of CHS cells.
An Improved Method for the Isolation of Eosinophilic Granulocytes From Peripheral Blood of Normal Individuals
Authors: Koenderman L , Kok P T M , Hamelink M L , Verhoeven A J , Bruijnzeel P L B 
Published: January 2018

Abstract
A simple and improved procedure is described for the isolation of human eosinophils from normal individuals with about 2% eosinophils in their peripheral blood. This method comprises a preincubation of a mixed granulocyte preparation with 10 nM fMLP for 10 min at 37° followed by a one-step density centrifugation on isotonic Percoll. The recovery of eosinophils is 49 ± 4% at 89 ± 4% purity. Because of the relatively high rate of recovery, it is now possible to isolate eosinophils from blood samples as small as 20 ml.
Because treatment with fMLP may alter the functional activity of the eosinophils, the following metabolic functions were tested: changes in cytosolic free Ca2+, oxygen consumption, chemiluminescence, chemotaxis, and leukotriene C4 formation. We found that 10 nM fMLP does not activate eosinophils in these assays, whereas 1 μM fMLP does (with the exception of chemotaxis). Furthermore, pretreatment of eosinophils with 10 nM fMLP did not influence the response to other stimuli in these assays. The usefulness of this method was evaluated by comparing it with three other previously described procedures. In our hands, only the method presented here enabled us to isolate eosinophils from normal individuals with about 2% eosinophils in their peripheral blood.