bluuebunny/crossref_metadata_2025_split huggingface dataset:
Palmitate-Derivatized Antibodies Can Specifically “Arm” Macrophage Effector Cells for ADCC
Authors: Colsky Arthur S , Peacock James S 
Published: January 2018

Abstract
The use of palmitate-derivatized antibodies (pal-Ab) for “arming” macrophage (Mφ) effector cells for antibody-dependent cellular cytotoxicity (ADCC) is described. Pal-Ab were incorporated onto the Mφ plasma membranes by insertion of the palmitate hydrocarbon chains into the outer leaflet of the phospholipid bilayer. Mφ bearing pal-Ab specific for chicken erythrocytes (CE) mediated efficient destruction of the CE targets. Neither non-ADCC effector cell populations nor pal-Ab consisting of antibody F(ab')2 fragments effected significant CE lysis. Mφ bearing pal-Ab that were not specific for CE did not mediate CE destruction, nor did anti-CE pal-Ab-bearing Mφ lyse nonspecific human erythrocyte targets. In this system of effector cell arming, the palmitate anchor of pal-Ab allows for the incorporation of large numbers of antibodies onto the effector cell surface, where they can promote efficient target cell capture and engage preexisting or newly expressed FcR on the effector cell surface. The results in this study, together with those from previous and ongoing investigations in which F(ab')2 pal-Ab were shown to mediate Fc receptor (FcR)-independent cytotoxicity by natural killer (NK) cells and Mφ populations at appropriate states of activation, suggest that pal-Ab, by directing both ADCC and FcR-independent effector cell activity onto a specified target, offer important advantages over other methods of effector cell arming.
Inhibition of Human Neutrophil Activation by the Allergic Mediator Release Inhibitor, CI-949
Authors: Wright Clifford D , Stewart Sheila F , Kuipers Paul J , Hoffman Michael D , Thueson David O , Conroy Mary Carol 
Published: January 2018

Abstract
The allergic mediator release inhibitor CI-949 [5-methoxy-3-(1-methylethoxy)-1-phenyl-N-1H-tetwol-5-yl-1H-indole-2-carboxamide, L-arginine salt] was evaluated for its effects on human neutrophil functions. CI-949 (100 μM) inhibited spontaneous migration and chemotaxis toward f-met-leu-phe (FMLP) by 49.1% and 45.8%, respectively. At the same concentration, CI-949 inhibited the phagocytosis of serum-opsonized zymosan (SOZ) by 39.0%. CI-949 inhibited leukotriene B4 and thromboxane B2 release in response to SOZ with IC50S of 2.0 μM and 3.3 μM, while inhibiting the response to FMLP with IC50s of 1.7 and 2.0 μM. CI-949 also inhibited myeloperoxidase release from primary lysosomal granules in response to the following stimuli with the respective IC50s (μM): C5a (40.3); FMLP (34.4): SOZ (21.4); concanavalin A (Con A) (3.9); and calcium ionophore A23187 (91.2). In contrast, CI-949 inhibited lysozyme release from secondary granules in response to SOZ and Con A with IC50S of 99.3 and 56.1 μM, while inhibiting the response to C5a, FMLP, and A23187 by 41.2%, 52.4%, and 10.0%, respectively, at 100 μM. CI-949 (100 μM) had no inhibitory effect against lysozyme release in response to L-α-1,2 dioctanoylglycerol (DiC8), or phorbol 12-myristate 13-acetate (PMA). CI-949 inhibited superoxide anion generation stimulated by FMLP and Con A with IC50S of 33.9 and 25.8 μM, while inhibiting the response to C5a, SOZ, and A23187 by 36.6%, 24.8%, and 14.1% and having no effect on the response to DIC8 or PMA at 100 μM. These results demonstrate preferential inhibition of arachidonic acid metabolism and degranulation of primary lysosomal granules by CI-949 with selectivity for stimuli which promote intracellular calcium mobilization or calcium influx.
Intracellular pH Regulates the Production of Different Oxygen Metabolites in Neutrophils: Effects of Organic Acids Produced by Anaerobic Bacteria
Authors: Tonetti Maurizio , Cavallero Annalisa , Botta Giuseppe A , Niederman Richard , Eftimiadi Costantino 
Published: January 2018

Abstract
The effects of short chain carboxylic acids (SCCA), namely succinic, butyric, and iso-butyric, on neutrophil metabolic activation were assessed. SCCA induced a significant decrease in O2- recovery and chemlluminescent response in neutrophils activated with the diacylglycerol analog tetradecanoyl-phorbol-acetate (TPA). SCCA did not alter O2 consumption, H2O2 production, or the calorimetrically determined energy expenditure occurring during the metabolic burst. SCCA also induced a significant acidification of intracellular pH (pHi). These results are consistent with an increased divalent versus univalent O2 reduction performed by the NADPH oxidase at a more acidic intracellular pH.
Gelatin Sponge Model of Effector Recruitment: Tumoricidal Activity of Adherent and Non-Adherent Lymphokine-Activated Killer Cells After Culture in interleukin-2
Authors: Akporiaye Emmanuel T , Barbieri Celia A , Stewart Carteton C , Bender James G 
Published: January 2018

Abstract
This study examined the specific tumoricidal activity of lymphokine-activated killer (LAK) cells derived from tumor-infiltrating lymphocytes that prevent the growth of secondary tumors in animals harboring progressing primary tumors. A pre-implanted gelatin sponge was employed to capture infiltrating host effectors during the expression of concomitant tumor immunity. Additionally, this study compared the cytolytic activity of these sponge-derived cells with those of counterpart splenic lymphocytes. The cells from both sources were cultured for 4 days in IL-2 to generate LAK cells which were further expanded in IL-2-containing medium for up to 11 days. The cytotoxic activities of these cells were measured in a Chromium-51 release assay. The data revealed that the culture of splenic, or sponge-derived lymphocytes results in the emergence of non-adherent and adherent cell populations with LAK activity. The 4-day sponge-derived LAK cells (adherent and non-adherent) exhibited significant cytotysis of EMT6 cells while the spleen-derived counterparts showed minimal cytotoxicity toward these targets. Some NK activity in LAK cells derived from both sources was evident by their lysis of YAC-1 cells. LAK cells from both sources were incapable of lysing histo-compatible EL-4 (H-2b) tumor cells. The lysis of the EMT6 cells by the sponge-derived LAK cells was maintained over an 11-day period of culture in IL-2. Conversely, the spleen-derived LAK cells were unable to significantly lyse EMT6 cells during this period of in vitro culture. These results show the superior specific tumoricidal activity of LAK cells derived from lymphocytes mediating tumor rejection in vivo (sponge-derived) over that of counterpart splenic lymphocytes.
Distinct Patterns of Chemotactic Peptide-Induced Calcium Mobilization in Differentiated Myeloid Leukemia Cells and Peripheral Blood Neutrophils
Authors: Laskin Debra L , Sirak Andrea A , Robertson Fredika M , Laskin Jeffrey D 
Published: January 2018

Abstract
Flow cytometry was used to compare intracellular calcium mobilization in mature neutrophilic granulocytes (PMN) with HL-60 promyelocytic leukemia cells induced to differentiate with dibutyryl cyclic adenosine 3’,5'-monophosphate (dbcAMP). Using the calcium-specific probe indo-1 acetoxymethyl ester, we found that the ability of differentiating HL-60 cells to mobilize calcium in response to N-formyl-methionyl-leucyl-phenylalanine (fMLP) developed concomitantly with expression of receptors on the cells for this peptide. Mobilization of calcium in HL-60 cells, as well as in PMN, in response to fMLP was dose dependent and was related to the presence of calcium in the culture medium. In calcium-free medium, tenfold higher concentrations of fMLP were required to induce calcium mobilization than in calcium-containing medium. In HL-60 cells, calcium mobilization occurred more rapidly than in PMN and was independent of extracellular calcium. Furthermore, the response of HL-60 cells to fMLP was more prolonged than the response of PMN and was also of greater magnitude. Calcium mobilization in both HL-60 cells and PMN was partially inhibited by the calcium channel blocker, verapamil, but completely blocked by trimethylbenzoic acid 8-(diethylamino) octyl ester, an intracellular calcium antagonist. These results indicate that although both cell types mobilize calcium in response to fMLP, the characteristics of the responses are distinct. These differences may underlie distinct functional responses of PMN and differentiated HL-60 cells to fMLP.
Activation of Macrophages From Aging Mice by Detoxified Lipid A
Authors: Chen Yifang , Solem Laura , Johnson Arthur G 
Published: January 2018

Abstract
A detoxified derivative of endotoxic lipopolysaccharides (LPS), monophosphoryl lipid A (MPL), which is capable of inducing nonspecific resistance against several infectious organisms, was tested for its capacity to activate peritoneal macrophages (Mϕ) from young and immunodeficient aging BALB/c and C3H/HeN mice. Superoxide generation and hydrogen peroxide release by Mϕ from aging mice were elevated following intraperitoneal injection with 25 µg of LPS or MPL, although they did not reach the peak levels achieved in LPS or MPL-treated young mice. Nitroblue tetrazolium reduction (NBT) by peritoneal Mϕ from aging C3H/HeN mice treated with MPL was higher than that in control aging mice, equalling that from MPL-treated young mice. LPS, its toxic counterpart, however, failed to increase NBT reduction in either group. MPL enhanced lysozyme activity in Mϕ from both aging and young C3H/HeN mice above initial control levels. On the other hand, LPS suppressed lysozyme activity in Mϕ from young, but not aging mice. Phagocytosis of Candida albicans by Mϕ from BALB/c mice was increased in both groups when stimulated by MPL, but not LPS. Similarly, MPL enhanced the ability to kill Candida in both aging and young BALB/c mice. This effect was not seen with LPS. Thus, a detoxified derivative of LPS was found capable of activating the respiratory burst, NBT reduction, elevating lysozyme activity, as well as phagocytosis and killing of Candida in murine peritoneal Mϕ from both young and aging mice.
Respiratory Burst Capability of Polymorphonuclear Neutrophils and TNF-α Serum Levels in Relationship to the Development of Septic Syndrome in Critically III Patients
Authors: Trautinger F , Hammerle A F , Pöschl G , Micksche M 
Published: January 2018

Abstract
Activated polymorphonuclear neutrophils (PMN) and neutrophil activating mediators such as tumor necrosis factor-α (TNF-α) are thought to be involved in the pathophysiology of sepsis and multiple organ failure syndrome (MOFS). In critically ill patients at high risk for the development of septic syndrome (n = 17) peripheral blood PMN were assayed for O-2 and H2O2 production after stimulation with phorbol myristate acetate (PMA, 40 nM). Serum TNF-α levels were determined by ELISA. At the time of admission to the intensive care unit we found significant higher levels of TNF-α (P = 0.0001) in the serum of patients finally developing sepsis correlating to higher respiratory burst capability in comparison to nonseptic patients. Additionally we were able to demonstrate a significant (P = 0.0016) lower dismutation rate of O-2 to H2O2 in deceased patients in comparison to survivors. These results give further evidence that elevated levels of circulating TNF-α and activated PMN play a significant role in the pathogenesis of septic syndrome in critically ill patients.
Macrophage Colony-Stimulating Factor Is Required for Human Monocyte Survival and Acts as a Cofactor for Their Terminal Differentiation to Macrophages In Vitro
Authors: Brugger Wolfram , Kreutz Marina , Andreesen Reinhard 
Published: January 2018

Abstract
Functional competence as well as phenotype heterogeneity of macrophages depend on the completion of their maturation pathway. Differentiation of committed myeloid progenitor cells is induced by colony-stimulating factors (CSF), but no consistent data exist on which factor(s) induce the terminal maturation from the circulating blood monocyte to the mature macrophage. In vitro, monocyte to macrophage transformation occurs in the presence of serum and can be followed by the expression of the maturation-associated antigens gp65-MAX.1, gp68-MAX.3, and CD51. We describe that the differentiation-inducing activity in serum cannot be replaced by any of the known and available purified recombinant cytokines. In the absence of serum monocytes die in suspension cultures while surviving as non-differentiating cells when cultured adherent to plastic. In serum-free suspension cultures survival can be significantly improved by the addition of recombinant human macrophage (rhM)-CSF whereas other cytokines do not. At any stage of serum-free adherent culture, monocyte to macrophage differentiation can be induced rapidly by the addition of serum, whereas cytokines (rhM-CSF, recombinant human granulocyte macrophage [rhGM]-CSF, recombinant human granulocyte [rhG]-CSF, recombinant human interleukin [rhIL]-1, rhIL-3, rhIL-4, rhIL-6, tumor necrosis factor [TNF]-α, interferon [IFN]-α, IFN-γ) alone or in combination are not effective. Serum-induced maturation, however, was suppressed in the presence of neutralizing anti-M-CSF antibodies. In addition to phenotype analysis, the secretory repertoire of rhM-CSF cultured monocytes was analyzed in comparison to serum cultured monocytes which further characterized them to be immature cells, i.e., low release of maturation-associated products such as α-2-macroglobulin, neopterin, fibronectin, and TNF-α, but high IL-6 secretion, an attribute of blood monocytes. We conclude that for monocyte survival in vitro the presence of endogenous M-CSF and possibly other autocrine factors elicited by cell adherence are required for the induction of macrophage maturation; however, yet undefined additional factor(s) are necessary. They are present in serum and may act in conjunction with M-CSF but are distinct from all known cytokines. Our in vitro system may be useful in the screening and discovery of these serum factor(s).
Differential Modulation of Surface Antigens on Human Macrophages by IFN-Gamma and GM-CSF: Effect on Susceptibility to LAK Lysis
Authors: Blanchard D Kay , Djeu Julie Y 
Published: January 2018

Abstract
We have previously reported that cultured human monocytes are lysed by autologous lymphokine-activated killer (LAK) cells in vitro and that treatment of monocytes with interferon-gamma (IFN-gamma) decreased their sensitivity to lysis. Conversely, incubation of monocytes with granulocyte-macrophage colony-stimulating factor (GM-CSF) significantly enhanced their susceptibility to LAK-mediated cytotoxicity. To determine if certain antigens were differentially modulated on macrophages by IFN-gamma and GM-CSF, cytokine-treated and untreated monocytes were analyzed for the expression of a variety of cell surface markers by flow cytometry. Cytotoxicity assays were performed to assess the ability of antibodies to each of these markers to block LAK lysis of macrophage target cells. While several of the surface structures were differentially modulated by cytokine treatment, it was found that only monoclonal antibodies to the adhesion proteins CD11a and CD18 were capable of blocking lysis of either cytokine-treated or untreated target macrophages.
Cryopreserved Cytoplasts From Human Polymorphonuclear Leukocytes (Cytokineplasts) Are Chemotactic at Speeds Comparable to Those of Fresh Intact Cells
Authors: Malawista Stephen E , de Boisfleury-Chevance Anne 
Published: January 2018

Abstract
Cytokineplasts (CKP) are granule-poor cytoplasts from human polymorphonuclear leukocytes (PMN) that retain motile function, even (unlike the parent PMN) after cryopreservation. Employing time-lapse videomicroscopy, we examined the chemotactic properties of CKP after cryopreservation toward erythrocytes lysed by laser microirradiation. Paths of locomotion were plotted for six CKP in the field, and velocities were calculated at 10-sec intervals. Mean velocities of the six fragments, ranging from 9.3 to 20.8 μm/min, are of the order of fresh, intact PMN, the fastest of locomoting cells.
Purification and Characterization of Macromolecular Phagocytosis Activators Released From Platelets
Authors: Sakamoto Haruhiko , Yokoya Yukihiro 
Published: January 2018

Abstract
Substantial amounts of macromolecular activators of phagocytosis from platelets (MAPPs) were released in response to exposure of platelets to the specific agonists thrombin and collagen and to calcium ionophore A23187. Centrifugation of the platelets in culture medium also resulted in a release of MAPPs, but not when the platelets were frozen and thawed prior to centrifugation.
In an experiment using outdated platelet concentrates, larger and smaller MAPPs (1-MAPP and s-MAPP, respectively) were purified from the thrombin stimulated supernatant (PRPr-plasma) by sequential steps of ammonium sulfate precipitation, delipidation with ethylacetate, ConA-Sepharose affinity chromatography, MONO Q anion exchange chromatography, and Superose 12 gel filtration. This procedure yielded 59,500-fold and 63,600-fold purified 1-MAPP (0.95 mg) and s-MAPP (0.41 mg), respectively, from 1,320 ml PRPr-plasma containing 84,500 mg protein. By gradient polyacrylamide gel electrophoresis the respective molecular weights (MW) of 1-MAPP and s-MAPP were 290–320 kd and 140–160 kd; isoelectric points were 5.6 and 4.6. Both MAPPs activated neutrophil phagocytosis of IgG-SRBC at concentrations in the range of 1 pM–100 nM. Indirect enzyme-linked immunosorbent assay (ELISA) and comparisons of the concentrations required for phagocytosis activation suggested that the MAPPs were not derived from fibronectin.
Involvement of HLA-DR+ Large Granular Lymphocytes in the Induction of Tumor Necrosis Factor by Mycobacterium avium-intracellulare Complex
Authors: Michelini-Norris M Beatriz , Blanchard D Kay , Friedman Herman , Djeu Julie Y 
Published: January 2018

Abstract
This study shows that normal human large granular lymphocytes (LGL) secrete tumor necrosis factor (TNF) in response to Mycobacterium avium-intracellulare complex (MAI). Percoll density gradient fractionation of peripheral mononuclear cells showed TNF activity in the fractions corresponding to LGL and not T cells, even when 5% monocytes were added to the T lymphocytes for accessory function. TNF release was not abrogated by treatment of the crude LGL preparations with anti-Leu M3. -CD4. and -CD8 antibodies (Ab) plus complement (C), but was abrogated by anti-CD16 and -CD2 Ab, as expected. Interestingly, anti-HLA-DR monoclonal antibody (mAb) treatment significantly diminished TNF activity from LGL, but maintained natural killer (NK) cell function unmodified as opposed to CD2 - and CD16- cell depletion. Panning studies demonstrated that TNF secretion upon MAI stimulation resided only in the HLA-DR+ LGL and not the DR LGL population. These results indicate that normal fresh HLA-DR+ LGL, as well as monocytes, are also responsible for rapid TNF secretion during early MAI infection. These DR - cells appear to be distinct from those expressing NK function.
Polyphosphoinositide Metabolism in Polymorphonuclear Cells From Healthy and Thermally Injured Rats: Effect of the Immunomodulator RU 41740
Authors: Tissot M , Mathieu J , Mirossay L , Thuret A , Giroud JP 
Published: January 2018

Abstract
Burn trauma is associated with alterations of various components of host defenses, including impaired neutrophil functions. In an animal model of experimental thermal injury, we studied if the modifications of cellular reactivity result from alterations in signalling systems by comparing polyphosphoinositide breakdown, particularly the production of inositol phosphates (IP, IP2 IP3), in healthy and burned rat polymorphonuclear neutrophil leukocytes (PMNs).
Neutrophil activators such as N-formyl-methionyl-leucyl-phenylalanine (fMLP) and serum-opsonized zymosan increased in vitro production of inositol phosphates in PMNs from healthy rats. The immunomodulator RU 41740 had no effect by itself, but decreased the stimulating effect of fMLP and zymosan. In PMNs from burned rats, the stimulating effects of fMLP and zymosan were decreased, while RU 41740 stimulated inositol phosphate generation. In vivo treatment with RU 41740 inhibited the activation of phosphoinositide metabolism by fMLP or zymosan in healthy rat PMNs. Similar treatment of burned rats after injury restored the stimulating effect of fMLP and zymosan on inositol phosphate accumulation in PMNs.
Thus, RU 41740 can modulate fMLP and zymosan receptor-mediated signal transduction, inducing an attenuation of the phosphatidylinositol hydrolysis response. After burn injury, when the activating effects of fMLP and zymosan are inhibited, RU 41740 can, on the contrary, stimulate phospholipase C-mediated polyphosphoinositide turnover and the formation of intracellular messengers such as IP3.
These data show that RU 41740 has different effects on polyphosphoinositide metabolism in rat PMNs, according to the physiological and pathological state of the animals. Interestingly, it has a beneficial action on the post-burn decrease in PMN reactivity.
Granule-associated flavin adenine dinucleotide (FAD) is responsible for eosinophil autofluorescence
Authors: Mayeno Arthur N , Hamann Kimm J , Gleich Gerald J 
Published: January 2018

Abstract
Unstained human eosinophils exhibit marked autofluorescence in comparison to other leukocytes due to a granule-associated fluorescent substance. Fluorescence spectroscopy of granule extracts reveals excitation maxima at ~380 and ~450 nm with a single emission at ~520, characteristic of flavins. The fluorescent material from eosinophil granule extracts was characterized by fluorescence, high-performance liquid chromatographic, and enzymatic analyses. First, acidification to pH 2.6 resulted in increased fluorescence, indicative of flavin adenine dinucleotide (FAD). Second, because flavin mononucleotide (FMN) and riboflavin cannot be distinguished by acidification, high-performance liquid chromatography was performed and revealed a predominance of FAD and smaller amounts (&lt;15%) of both FMN and riboflavin. Third, the presence of FAD was clearly demonstrated by reconstitution of the activity of d-amino acid oxidase, a FAD-dependent enzyme, when granule extracts were added to the apoenzyme. Thus, we have identified FAD as the predominant fluorophore in eosinophil granules. The small amounts of FMN and riboflavin detected may result from the hydrolysis of FAD under the acidic conditions of granule extraction. Because fluorescent material is deposited onto target cells by eosinophils, it is possible that granule-associated flavoproteins may act as a source of hydrogen peroxide and/or superoxide, which, in conjunction with eosinophil peroxidase, could yield potent cytotoxic agents.
Effects of Pseudomonas aeruginosa rhamnolipids on human monocyte-derived macrophages
Authors: McClure Carol D , Schiller Neal L 
Published: January 2018

Abstract
Pseudomonas aeruginosa, a major opportunistic gram-negative pathogen, produces and secretes two heat-stable hemolytic glycolipids, a monorhamnolipid and a dirhamnolipid. In this paper a simplified method for the isolation of these rhamnolipids is described. The effect of these two rhamnolipids, both together and individually, on the viability and structural morphology of human monocyte-derived macrophages (MDMs) was examined. These cells were found to be very susceptible to the cytolytic activity of the rhamnolipids, particularly the dirhamnolipid. The monorhamnolipid, although not as cytolytic as the dirhamnolipid, caused extensive blebbing of the MDM plasma membrane. Comparison studies with several detergents confirmed the different yet distinct detergent-like activity of each rhamnolipid form. At sublethal doses, the rhamnolipids produced marked cellular distortions of the MDMs and inhibited the ability of these cells to bind and/or ingest preopsonized bacteria. The potential mechanism of action of these rhamnolipids on the MDM membranes is discussed, as well as the possible significance of these extracellular bacterial glycolipids as a virulence factor in the pathogenesis of P. aeruginosa.