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1.
Ann Surg ; 210(3): 355-72; discussion 372-3, 1989 Sep.
Article in English | MEDLINE | ID: mdl-2549891

ABSTRACT

Thirty multiply injured blunt-trauma patients at high risk for development of ARDS (multisystem trauma including more than one organ or extremity, Injury Severity Score of 26 or more, hypotension and need for 1500 mL or more blood within the first hour after admission, and PaO2 less than or equal to 70 torr) were studied sequentially with blood and physiologic evaluations beginning immediately after injury and every eight hours for eight days, or until death, to study the evolution of the ARDS process. Mixed venous blood samples were obtained for eicosanoids PGE2, PGF2 alpha, thromboxane B2, PGI2 (6-KetoPGF1 alpha) and leukotriene B4 (LTB4). Platelet (PLAT), and neutrophil (WBC) counts were also done and plasma elastase was measured. At 7:00 AM each day patient neutrophils were obtained for a study of zymosan-activated superoxide production using a chemiluminescence assay. These data were correlated with physiologic measurements of the Respiratory Index (RI), per cent pulmonary shunt (QS/QT), and respiratory compliance measures. Seven patients developed a fulminant post-traumatic ARDS syndrome within 96 hours after injury. Twelve patients without ARDS developed sepsis (TS) four or more days after injury, and 11 had uncomplicated postinjury courses (TR). Compared to both TR and TS, ARDS had a significant (p less than 0.01) rise in neutrophil superoxide production beginning on day 2 through day 4 after injury. This was preceded by rises in PGE2 and LTB4, which were significantly correlated with subsequent falls in PLAT and WBC and rises in TXB2, PGF1, and superoxide production and followed by increases in RI, QS/QT, and a fall in compliance. The significant difference in the pattern and sequence of events in ARDS compared to TR and TS patients suggests that in ARDS the earliest event may be related to peripheral release of PGE2 and LTB4 due to platelet activation and lung sequestration with release of PGF2 alpha, and by aggregation and leukocyte adherence with release of elastase. However, fulminant ARDS mortality appears to be related to the subsequent amplification of the LTB4 leukocyte activation with superoxide production that does not achieve significance before the second day after injury and rises to a maximum by day 4 after injury. These data suggest that post-trauma ARDS follows a different evolutionary pattern than that reported in animal models and is also different from that seen in human TS or TR patients.(ABSTRACT TRUNCATED AT 400 WORDS)


Subject(s)
Blood Platelets/physiology , Neutrophils/physiology , Prostaglandins/physiology , Respiratory Distress Syndrome/etiology , Wounds and Injuries/complications , Biomechanical Phenomena , Cell Communication , Humans , Infections/etiology , Infections/physiopathology , Leukotriene B4/physiology , Respiratory Distress Syndrome/physiopathology , Statistics as Topic , Superoxides/physiology , Thromboxane B2/physiology , Time Factors , Wounds and Injuries/physiopathology
2.
Cell ; 58(5): 923-31, 1989 Sep 08.
Article in English | MEDLINE | ID: mdl-2476237

ABSTRACT

Tumor necrosis factor (TNF) induces the synthesis of protein(s) that can protect cells against subsequent killing by TNF in the presence of cycloheximide. Here we demonstrate that manganous superoxide dismutase (MnSOD), a mitochondrial enzyme involved in the scavenging of superoxide radicals (O2-), is such a protein. Overexpression of MnSOD confers increased resistance to TNF plus cycloheximide on the 293 human embryonic kidney cell line. Conversely, expression of antisense MnSOD RNA renders these cells sensitive to TNF even in the absence of cycloheximide. The TNF sensitivity of the ME-180 human cervical carcinoma cell line can also be modulated through expression of sense and antisense MnSOD RNAs. These data identify MnSOD as an important determinant of cellular resistance to TNF and implicate mitochondrially generated O2- as a key component of TNF-mediated tumor cell killing.


Subject(s)
Superoxide Dismutase/physiology , Superoxides/physiology , Tumor Necrosis Factor-alpha/toxicity , Cell Line , Humans , In Vitro Techniques , Manganese , Mitochondria/physiology , RNA/pharmacology , RNA, Antisense , Transfection
3.
J Biol Chem ; 264(16): 9243-9, 1989 Jun 05.
Article in English | MEDLINE | ID: mdl-2542303

ABSTRACT

Novel metal complexes, Fe(II)-tetrakis-N,N,N',N' (2-pyridylmethyl)ethylenediamine(Fe-TPEN) and Fe(III)-tris[N-(2-pyridylmethyl)-2-aminoethyl]amine (Fe-TPAA), catalyzed the dismutation of superoxide, and 0.8 microM Fe-TPEN and 7.5 microM Fe-TPAA were equivalent to 1 unit of superoxide dismutase (SOD) activity in the xanthine oxidase-cytochrome c assay. Addition of serum albumin had no effect on the activities of Fe-TPEN and Fe-TPAA but depressed those of the Cu(salicylate)2 and Cu(diisopropylsalicylate)2 complexes. Both iron complexes blocked the toxic effect of paraquat on Escherichia coli growth and survival without causing induction of SOD. In contrast, this behavior was not seen with other SOD mimics containing copper or manganese. These results support the view that the SOD activities of these iron complexes remain intact in living cells.


Subject(s)
Escherichia coli/enzymology , Iron , Superoxide Dismutase/metabolism , Chelating Agents , Cytochrome c Group/metabolism , Enzyme Induction/drug effects , Escherichia coli/drug effects , Escherichia coli/growth & development , Ethylenediamines , Oxidation-Reduction , Paraquat/toxicity , Superoxide Dismutase/biosynthesis , Superoxides/physiology , Xanthine Oxidase/metabolism
4.
Biochem Biophys Res Commun ; 161(2): 762-70, 1989 Jun 15.
Article in English | MEDLINE | ID: mdl-2544172

ABSTRACT

Superoxide anion radical and superoxide dismutase, the enzyme responsible for dismutating it, are both present in the ovary and uterus of Mus musculus during early pregnancy. The detectable, stable levels of superoxide radical and the constant high levels of superoxide dismutase in the ovary during early pregnancy suggests that these may be involved in the regulation of extended luteal steroidogenesis for the maintenance of pregnancy. An inverse correlation between the levels of superoxide anion radical and superoxide dismutase in the uterus is shown. The high levels of superoxide anion radical in the uterus on the early morning of Day 5 of pregnancy point towards a probable role for this radical in the act of implantation and in mediating the increased vascular permeability at the initiation of implantation.


Subject(s)
Blastocyst/physiology , Embryo Implantation , Superoxides/physiology , Animals , Capillary Permeability , Electron Spin Resonance Spectroscopy , Female , Mice , Ovary/metabolism , Superoxide Dismutase/metabolism , Time Factors , Uterus/metabolism
5.
Biochem Biophys Res Commun ; 161(2): 771-5, 1989 Jun 15.
Article in English | MEDLINE | ID: mdl-2544173

ABSTRACT

Mammalian spermatozoa possess a membrane-located 'proton pump' which appears to regulate the acquisition and maintenance of their motility. The 'proton pump' seems to be coupled with the superoxide radical-superoxide dismutase system which, when uncoupled, results in total loss of sperm motility. Addition of sulfhydryl-compounds restarts the proton pump and reactivates sperm motility. Cobaltous ions block the membrane sulfhydryl groups and it is proposed that the sulfhydryl groups may be involved in regulating proton secretion and sperm motility.


Subject(s)
Hydrogen-Ion Concentration , Sperm Motility , Spermatozoa/physiology , Superoxide Dismutase/metabolism , Superoxides/physiology , Animals , Biological Transport, Active , Cobalt/pharmacology , Cysteine/pharmacology , Male , Rats , Spermatogenesis
6.
J Clin Invest ; 83(4): 1326-35, 1989 Apr.
Article in English | MEDLINE | ID: mdl-2467923

ABSTRACT

We have developed a model of reperfusion injury in Krebs buffer-perfused rabbit lungs, characterized by pulmonary vasoconstriction, microvascular injury, and marked lung edema formation. During reperfusion there was a threefold increase in lung superoxide anion (O2-) production, as measured by in vivo reduction of nitroblue tetrazolium, and a twofold increase in the release of O2- into lung perfusate, as measured by reduction of succinylated ferricytochrome c. Injury could be prevented by the xanthine oxidase inhibitor allopurinol, the O2- scavenger SOD, the hydrogen peroxide scavenger catalase, the iron chelator deferoxamine, or the thiols dimethylthiourea or N-acetylcysteine. The protective effect of SOD could be abolished by the anion channel blocker 4,4'-diisothiocyano-2,2'-stilbene disulfonic acid, indicating that SOD consumes O2- in the extracellular medium, thereby creating a concentration gradient favorable for rapid diffusion of O2- out of cells. Our results extend information about the mechanisms of reperfusion lung injury that have been assembled by studies in other organs, and offer potential strategies for improved organ preservation, for treatment of reperfusion injury after pulmonary thromboembolectomy, and for explanation and therapy of many complications of pulmonary embolism.


Subject(s)
Lung Diseases/pathology , Reperfusion Injury/metabolism , Superoxide Dismutase/physiology , Superoxides/physiology , 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid , 4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid/analogs & derivatives , Animals , Antioxidants/pharmacology , Cytochrome c Group/metabolism , Disease Models, Animal , Free Radicals , Ion Channels/drug effects , Ion Channels/metabolism , Lung Diseases/metabolism , Male , Nitroblue Tetrazolium/metabolism , Rabbits , Reperfusion Injury/enzymology , Xanthine Oxidase/antagonists & inhibitors
7.
J Immunol ; 142(6): 1963-9, 1989 Mar 15.
Article in English | MEDLINE | ID: mdl-2537865

ABSTRACT

Human monocytes, upon activation with opsonized zymosan, altered low-density lipoprotein (LDL) during a 24-h co-incubation, resulting in its oxidation and acquisition of cytotoxic activity against target fibroblast cell lines. Both the oxidation of LDL and its conversion to a cytotoxin were enhanced with time of incubation, with the most substantial changes occurring after 6 h of culture of LDL with activated monocytes. Unactivated monocytes did not mediate either alteration. Superoxide anion (O2-) participated in both the oxidation of LDL and its conversion to a cytotoxin since addition of superoxide dismutase (SOD) at the beginning of the co-incubation inhibited, in a concentration dependent fashion, both the monocyte-mediated oxidation and the monocyte-mediated conversion of LDL to a cytotoxin. As expected, the rate of superoxide anion release was greatest during the respiratory burst, very early in the 24-h incubation (0 to 2 h); however, exposure of LDL to monocytes during the respiratory burst was not required for LDL oxidation. The lower levels of O2- released by the cells hours after the respiratory burst had subsided were sufficient to lead to the initiation of LDL oxidation. Three results indicated that the oxidative modification of LDL into a cytotoxin required O2(-)-independent free radical propagation after O2(-)-dependent initiation. First, oxidation of LDL exposed to the activated, superoxide anion-releasing monocytes for 6 h could be almost completely blocked by the addition at 6 h of the general free radical scavenger butylated hydroxytoluene, but not by SOD. Second, LDL oxidation proceeded even after removal of LDL from the superoxide anion-producing, activated cells after various durations of exposure. Third, the development of substantial levels of lipid peroxidation products and the development of greater cytotoxicity occurred after 6 h of exposure of LDL to activated cells, long after peak O2- release had subsided. These results lead us to conclude that monocyte-mediated oxidation of LDL, leading to its transformation into a cytotoxin, requires release of O2- occurring as a result of activation but not necessarily during the respiratory burst, and also requires O2(-)-independent free radical propagation. The modification of LDL into a potent toxin by activated monocytes may explain the tissue damage in atherosclerotic lesions and other pathologic sites in which inflammatory cells congregate.


Subject(s)
Cytotoxins/metabolism , Lipid Peroxidation , Lipoproteins, LDL/metabolism , Monocytes/physiology , Superoxides/physiology , Free Radicals , Humans , Lipid Peroxidation/drug effects , Macrophage Activation/drug effects , Monocytes/drug effects , Superoxide Dismutase/pharmacology , Thiobarbiturates
8.
Biochem Biophys Res Commun ; 159(2): 584-91, 1989 Mar 15.
Article in English | MEDLINE | ID: mdl-2539116

ABSTRACT

Unstimulated alveolar macrophages, but not polymorphonuclear leukocytes, elicit chemiluminescence from lucigenin which cannot be entirely accounted for by the resting level of superoxide generation. This chemiluminescence was inhibited by both superoxide scavengers and inhibitors of mitochondrial respiration. Although 12-O-tetradecanoyl phorbol-13-acetate addition resulted in a significant increase in cellular superoxide generation, an unexpected decrease in lucigenin chemiluminescence was noted. These results suggest that mitochondria in alveolar macrophages may be a site of lucigenin accumulation and dioxygenation and that 12-O-tetradecanoyl phorbol-13-acetate may modulate this activity.


Subject(s)
Acridines , Luminescent Measurements , Macrophages/metabolism , Mitochondria/metabolism , Oxygen Consumption , Animals , Macrophages/drug effects , Male , Oxygen Consumption/drug effects , Pulmonary Alveoli , Rats , Rats, Inbred Strains , Submitochondrial Particles/drug effects , Submitochondrial Particles/metabolism , Superoxides/biosynthesis , Superoxides/physiology
9.
J Immunol ; 142(4): 1260-7, 1989 Feb 15.
Article in English | MEDLINE | ID: mdl-2536769

ABSTRACT

EBV-transformed B lymphocyte cell lines can generate superoxide, using an electron transport chain homologous, or even identical, to phagocytic NADPH-oxidase. We searched for normal, not virally transformed, B lymphocytes with analogous properties, using tonsils as the source of B cells. Unseparated tonsillar leukocytes contained cells capable of PMA-triggered superoxide dismutase-inhibitable reduction of nitroblue tetrazolium (NBT+ cells) well in excess of phagocytes (18.9 +/- 6.4% NBT+ cells with 1.3 +/- 0.9% granulocytes and 1.9 +/- 2.3% monocytes/macrophages, n = 8). NBT reduction was also inhibited by diphenylene iodonium, a selective inhibitor of phagocytic NADPH-oxidase. Cross-linking of surface Ig was equally effective as PMA in inducing NBT reduction among tonsillar leukocytes. NBT+ cells co-distributed with B cells on Percoll density gradients and were enriched among purified B cells obtained by SRBC rosetting twice and Sephadex G10 adherence (47.8 +/- 15.2% NBT+ cells among 90.5 +/- 5.5% B cells, 4.8 +/- 5.1% T cells, 1.2 +/- 0.77% monocytes/macrophages, and 0.73 +/- 0.6% granulocytes, n = 10). Further, mAb 7D5, directed against an extracellularly located epitope of the small subunit of cytochrome b-245 of phagocytes, stained the majority of tonsillar B cells (85 +/- 9.2% 7D5+ cells and 91.6 +/- 4.04% B cells, n = 3). Superoxide production, staining with 7D5 antibody, and expression of mRNA for the beta chain of cytochrome b-245 were further analyzed in cell lines. The EBV-BLCL F1 and the Burkitt lymphoma P3HR-1 both carried 7D5-detectable cytochrome b-245 Ag and expressed mRNA for the beta chain of the cytochrome b, both in similar amounts. However, only F1, not P3HR-1, was capable of PMA-triggered superoxide production. These data indicate that also normal nontransformed B lymphocytes possess the capacity to generate superoxide by a system apparently similar to phagocytic NADPH-oxidase, provisionally termed "B cell oxidase." Discrepancies observed in certain B cells and lines between expression of cytochrome b components and stimulus-induced superoxide production may be related to an absence or low level of other oxidase components or of the signal transduction mechanism. Conceivably, production of superoxide and derived reactive oxygen species by B cells may have cytotoxic, immunomodulatory, or mutagenic effects on the B cells themselves or on cells in their immediate vicinity.


Subject(s)
B-Lymphocytes/metabolism , Cytochrome b Group/metabolism , Nitroblue Tetrazolium , Palatine Tonsil/metabolism , Superoxides/physiology , Tetrazolium Salts , Adult , Antibodies, Monoclonal , Antigens, Surface/analysis , B-Lymphocytes/classification , Cell Line , Child , Cytochrome b Group/immunology , Humans , Oxidation-Reduction , Phenotype , RNA, Messenger/isolation & purification , Superoxides/biosynthesis
10.
Free Radic Res Commun ; 5(6): 315-8, 1989.
Article in English | MEDLINE | ID: mdl-2538380

ABSTRACT

It is proposed that vascular endothelium has an intrinsic capacity to generate O2- for regulatory purposes such as inactivation of endothelium-derived relaxing factor. Ischaemia can disrupt the functioning of this oxidant-generating system, resulting in greater O2- generation when O2 is restored. Ischaemia-induced cellular injury can also lead to release of iron ions, that, upon reperfusion, cause conversion of O2- and H2O2 to powerfully-oxidizing species (such as .OH) that further injure the endothelium.


Subject(s)
Endothelium, Vascular/physiology , Iron/physiology , Reperfusion Injury , Superoxides/physiology , Animals , Humans , Superoxide Dismutase/physiology
12.
Blood ; 72(1): 273-81, 1988 Jul.
Article in English | MEDLINE | ID: mdl-2839254

ABSTRACT

To better understand the limited hematopoietic life span of human marrow "Dexter" cultures, we developed a miniaturized, two-stage culture system with which in vitro production of hematopoietic progenitors could be reproducibly detected and quantified. Light-density, gradient-separated human marrow cells were inoculated into Leighton slide tubes, and adherent ("stromal") cell layers were allowed to develop on the removable coverslips within these tubes during an initial 4 weeks of culture. Once stromal cell layers were established, cultures were irradiated (800 cGy) to eliminate all residual hematopoietic progenitors. The cultures were then recharged with autologous, cryopreserved marrow cells (enriched for BFU-E and CFU-GM) to reconstitute stem cell populations and to initiate in vitro hematopoiesis. Most progenitor cells added to irradiated cultures were no longer detectable by clonal assays within one to four days after recharge. Nonetheless, stable populations of adherent BFU-E and CFU-GM became established in these cultures within 24 to 48 hours, and when the total numbers of progenitors (adherent and nonadherent) were measured at weekly intervals thereafter, it was evident that both BFU-E and CFU-GM were generated in vitro. However, progenitor cell production declined as neutrophils and macrophages accumulated in the cultures. Moreover, with this accumulation of mature myeloid cells, increasing levels of O2- and H2O2 could be detected in the cultures, and it was found that the addition of oxidant scavengers (catalase and mannitol) to culture media enhanced the weekly expansions of progenitor cell numbers that could be measured. These findings support the conclusion that reactive O2 intermediates generated by mature myeloid cells have a role in limiting the duration and extent of hematopoietic progenitor cell self-renewal in long-term "Dexter" cultures of human marrow.


Subject(s)
Bone Marrow Cells , Culture Techniques/methods , Hematopoiesis , Hematopoietic Stem Cells/cytology , Oxygen Consumption , Cells, Cultured , Free Radicals , Hematopoietic Stem Cells/metabolism , Hematopoietic Stem Cells/physiology , Humans , Hydrogen Peroxide/biosynthesis , Hydrogen Peroxide/physiology , Superoxide Dismutase/pharmacology , Superoxides/biosynthesis , Superoxides/physiology , Time Factors
13.
J Invest Dermatol ; 90(5): 652-7, 1988 May.
Article in English | MEDLINE | ID: mdl-2834456

ABSTRACT

Photoradiation therapy with porphyrins and light offers an alternative approach to the management of certain types of cancer. The mechanism of tissue destruction mediated by this modality is poorly understood. In this study, epidermal microsomes incubated in vitro with Photofrin-I (Pf-I) and Photofrin-II (Pf-II) followed by exposure to radiation (approximately 400 nm) resulted in increased (180%) NADPH-supported (enzymatic) as well as ADP/iron-supported (140%) (nonenzymatic) lipid peroxidative damage as measured by malondialdehyde formation. Lipid peroxidation by Pf-I and Pf-II was found to be differentially affected by quenchers of singlet oxygen (2,5-dimethylfuran, histidine, beta-carotene, ascorbic acid, and sodium azide), superoxide anion (superoxide dismutase), and the hydroxyl radical (sodium benzoate, mannitol, and ethanol). Catalase, a quencher of hydrogen peroxide, afforded significant protection only against Pf-II-enhanced lipid peroxidative damage while it had little effect against the Pf-I-mediated reaction. Deuterium oxide, which is known to increase the half-life of singlet oxygen, was found to enhance Pf-I-mediated lipid peroxidation but produced insignificant effects upon Pf-II-mediated photosensitization. Our results indicate that Pf-I and Pf-II, which are employed for the photodynamic therapy of malignant tumors, evoke membrane damage by generating different reactive oxygen species. The Pf-I-mediated photodestruction mainly involves a type II mechanism via singlet oxygen formation, whereas Pf-II-mediated photodestruction preferentially involves a type I mechanism by generating superoxide anions and hydroxyl radicals. Our data indicate that tumor necrosis evoked by porphyrins and light is likely due to the generation of reactive oxygen species.


Subject(s)
Epidermis/metabolism , Hematoporphyrins/pharmacology , Lipid Peroxides/biosynthesis , Oxygen/physiology , Radiation-Sensitizing Agents/pharmacology , Animals , Anions/physiology , Dihematoporphyrin Ether , Epidermis/ultrastructure , Hematoporphyrin Derivative , Hydrogen Peroxide/physiology , Hydroxides/physiology , Hydroxyl Radical , Intracellular Membranes/metabolism , Microsomes/metabolism , Microsomes/ultrastructure , Oxygen/metabolism , Superoxides/physiology
14.
Baillieres Clin Rheumatol ; 2(1): 211-32, 1988 Apr.
Article in English | MEDLINE | ID: mdl-2843297

ABSTRACT

There is evidence for both oxygen-centred free radicals and products of complement activation acting as mediators of inflammation. Evidence for the generation and reaction of free radicals at sites of inflammation can only be indirect and circumstantial due to their very transient nature. Evidence for complement activation in several inflammatory conditions, including rheumatoid arthritis is strong. These mediator classes individually possess a range of potential proinflammatory activities. Their effects may be linked through the formation of immune complexes and the activation of polymorphonuclear leukocytes. Their actions will also be linked with and modulated by the activities of other mediators mentioned only briefly in this chapter. The relative importance of the different mediators in any particular inflammatory condition is difficult to ascertain. The importance of free radicals and complement will be better understood when drugs specifically and unequivocally aimed at their control are identified. This potential for therapeutic advances in the treatment of inflammatory disorders has yet to be realized.


Subject(s)
Arthritis/etiology , Complement System Proteins/physiology , Hydroxides/physiology , Immunoglobulins/physiology , Superoxides/physiology , Animals , Antigen-Antibody Complex/physiology , Arthritis/drug therapy , Arthritis/immunology , Arthritis, Rheumatoid/metabolism , Free Radicals , Humans
15.
Scand J Rheumatol Suppl ; 75: 148-56, 1988.
Article in English | MEDLINE | ID: mdl-2853449

ABSTRACT

UNLABELLED: Immunoglobulin G from rheumatoid patients is denatured around the hinge region. This has been proposed as an explanation for the presence of circulating autoantibodies to IgG in these patients. It has previously been suggested that oxygen radicals (OR) derived from activated polymorphs may play a role in denaturation in vivo. Using sera from rheumatoid patients and age-matched controls in a modified ELISA technique, we have investigated the potential for polyclonal rheumatoid factors (RF) to bind to OR denatured IgG. Three model systems were used to generate OR in vitro: (a) purified PMN s activated by the cell surface stimulant PMA, (b) radiolysis of IgG in solution to generate specifically the superoxide radical and, in a separate system, the hydroxyl radical, (OH.), (c) purified myeloperoxide in the presence of H2O2 and halide ions. RESULTS: 1. The binding of both IgA and IgM RF s to PMN denatured IgG increased dose dependently for seropositive sera only. 2. The OH. radical but not the superoxide radical significantly increased the binding of IgA and M RF, again only for seropositive sera. 3. The myeloperoxidase enzyme system did not increase RF binding. 4. IgG incubated with elastase was not found to be a better antigen than native IgG. These results indicate that IgG is denatured by OR released from activated PMN, thereby producing an antigen for polyclonal RF s.


Subject(s)
Immunoglobulin G/immunology , Neutrophils/metabolism , Oxygen/physiology , Rheumatoid Factor/immunology , Arthritis, Rheumatoid/blood , Enzyme-Linked Immunosorbent Assay , Free Radicals , Gamma Rays , Humans , Oxygen/metabolism , Superoxides/physiology
17.
J Appl Physiol (1985) ; 63(4): 1526-32, 1987 Oct.
Article in English | MEDLINE | ID: mdl-2826378

ABSTRACT

We hypothesized that superoxide anion (O2-.) and hydrogen peroxide (H2O2) might be important mediators of endotoxin-induced acute respiratory failure (ARF) in pigs. As specific scavengers of O2-. and H2O2, we infused polyethylene glycol-superoxide dismutase (PEG-SOD; 2,000 IU/kg) and PEG-catalase (CAT; 15,000 IU/kg), respectively. Escherichia coli endotoxin (055-B5) was infused intravenously into anesthetized 10- to 14-wk-old pigs at 5 micrograms/kg the 1st h, followed by 2 micrograms.kg-1.h-1 for 3.5 h. During phase 1 (i.e., 0-2 h) and 2 (i.e., 2-4.5 h), endotoxin decreased cardiac index (CI) and lung dynamic compliance, and increased mean pulmonary arterial pressure (Ppa), pulmonary vascular resistance (PVR), total peripheral resistance (TPR), alveolar-arterial O2 gradient, and hematocrit. Endotoxemia also caused granulocytopenia and increased the postmortem bronchoalveolar lavage fluid (BALF) albumin concentration and wet-to-dry ratio of bloodless lung. During endotoxemia, PEG-SOD failed to significantly alter any measured or calculated parameter. On the other hand, PEG-CAT attenuated the early (i.e., 0-1 h) endotoxin-induced decrease in CI and increases in Ppa, PVR, and TPR, but failed to modify these parameters during phase 2. PEG-CAT also attenuated the endotoxin-induced granulocytopenia and the increased BALF albumin concentration. In the presence of inactivated PEG-CAT, these protective effects were reversed. We conclude that O2-. does not directly contribute to endotoxin-induced lung injury and that H2O2 (or a subsequent metabolite) contributes to the early endotoxin-induced hemodynamic changes, granulocytopenia, and increased permeability of the alveolar-capillary membrane.


Subject(s)
Catalase/pharmacology , Endotoxins/blood , Respiratory Insufficiency/prevention & control , Superoxide Dismutase/pharmacology , Animals , Enzymes, Immobilized/pharmacology , Free Radicals , Hydrogen Peroxide/physiology , Polyethylene Glycols , Respiratory Insufficiency/etiology , Superoxides/physiology , Swine
18.
J Immunol ; 139(4): 1342-7, 1987 Aug 15.
Article in English | MEDLINE | ID: mdl-3039002

ABSTRACT

Activation of human peripheral blood monocytes results in the expression of interleukin 2 (IL 2) receptors, which are absent on resting monocytes. In a population of purified monocytes, the appearance of IL 2 receptors occurs on the majority of cells in association with increased levels of HLA-DR. Lipopolysaccharide (LPS) induces maximum numbers of IL 2 receptors within 12 hr, whereas IFN-gamma requires 48 hr. We used cDNA encoding for the human IL 2 receptor to evaluate IL 2 receptor gene expression in resting and activated monocytes. Within 4 hr after LPS stimulation, IL 2 receptor mRNA species of 3500 and 1500 bases appear, reaching peak levels between 8 and 12 hr and declining thereafter. The LPS-activated monocyte IL 2 receptor protein is expressed on the cell surface within a few hours after the detection of IL 2 receptor mRNA. The addition of IL 2 to IL 2 receptor-positive monocytes augments their generation of reactive oxygen intermediates and their cytotoxic activity. Thus monocytes when activated undergo a series of morphologic, phenotypic, and functional changes, including the expression of IL 2 receptors, which may provide an important immunoregulatory pathway.


Subject(s)
Interleukin-2/physiology , Monocytes/physiology , Receptors, Immunologic/genetics , Acetylmuramyl-Alanyl-Isoglutamine/pharmacology , Cytotoxicity, Immunologic , Gene Expression Regulation , Giardia/immunology , HLA-DR Antigens/immunology , Humans , Immunity, Cellular , Macrophage Activation , N-Formylmethionine Leucyl-Phenylalanine/pharmacology , RNA, Messenger/genetics , Receptors, Interleukin-2 , Superoxides/physiology , Tetradecanoylphorbol Acetate/pharmacology
20.
Cancer Res ; 46(11): 5571-5, 1986 Nov.
Article in English | MEDLINE | ID: mdl-3019529

ABSTRACT

Superoxide dismutase mimetic copper(II) complexes, such as copper(II)(3,5-diisopropylsalicylate)2 (CuDIPS), inhibit phorbol ester stimulated tumor promotion in mouse skin. Therefore, CuDIPS was tested as a potential inhibitor of another effect of phorbol esters, induction of interleukin 2 (IL2) synthesis, in the mouse thymoma cell line EL4. CuDIPS inhibited phorbol ester induced IL2 production in a concentration dependent manner with a 50% inhibitory concentration of about 10 microM. However, the ligand 3,5-diisopropylsalicylic acid also inhibited the induction of IL2 by phorbol esters (50% inhibitory concentration, 15 microM). Since the superoxide dismutase mimetic activity of CuDIPS is not stable in the presence of ethylenediaminetetraacetic acid, the effects of CuDIPS could be due to the free ligand and not to the intact metallocomplex. Consequently, a series of extremely stable copper(II) macrocyclic compounds was synthesized, and the reduction potential, superoxide dismutase mimetic activity, and ability to inhibit phorbol ester induced IL2 production were determined for each. Of the copper(II) macrocyclic complexes studied, only the most potent superoxide dismutase mimetic compound was found to inhibit phorbol ester induced IL2 production. Copper(II) complexes had to be added no later than 4 following phorbol ester administration to be effective inhibitors of the IL response, suggesting that these compounds act subsequent to the binding of phorbol esters but prior to the transcription of IL2 messenger RNA. Adherence of EL4 cells to substrate in response to phorbol esters was unaffected by copper(II) compounds. In summary, copper(II) compounds with appropriate reduction potentials can act within a defined time period to inhibit some, but not all, of the effects of phorbol esters on EL4 cells.


Subject(s)
Copper/pharmacology , Interleukin-2/biosynthesis , Phorbol Esters/antagonists & inhibitors , Salicylates/pharmacology , Superoxides/physiology , Animals , Cations, Divalent , Cell Adhesion/drug effects , Mice , Protein Biosynthesis , Solubility , Superoxide Dismutase/metabolism , Time Factors
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