leukotriene-b4 and Leukopenia

Leukotriene B4 (LTB4) plays an important role in acute and chronic inflammatory and hypersensitive reactions. We studied the time course of LTB4 biosynthesis in whole blood in 18 patients with end-stage renal failure maintained on regular hemodialysis with two different membranes, cuprophane and polyacrylonitrile (AN 69). The basal levels of LTB4 from dialysis patients did not differ significantly from a normal control group. Compared to predialytic values, the cuprophane membrane caused a maximal release of LTB4 by a factor of about 4.5 (p less than 0.01) within the first 10 to 20 minutes. Thereafter the level fell and returned to baseline range at the end of the hemodialysis session. With the use of the AN 69 membrane no significant increase of LTB4 could be demonstrated. The changes in LTB4 concentration showed a close temporal correlation to the alterations in white blood cell count. We conclude that (1) LTB4 is a biologically important mediator of neutrophil activation during hemodialysis, and (2) LTB4 may be a sensitive marker of biocompatibility in vivo.

Topics: Acrylic Resins; Biocompatible Materials; Cellulose; Female; Humans; Kidney Failure, Chronic; Kidneys, Artificial; Leukopenia; Leukotriene B4; Male; Membranes, Artificial

The interference of the 5-lipoxygenase inhibitor, BW B70C ((E)-N-(3-[3-(4-fluorophenoxy)phenyl]-1(R,S)-methyl prop-2-enyl)-N-hydroxyurea), with Escherichia coli lipopolysaccharide (endotoxin)-induced lung leucocyte sequestration and microvascular albumin exchanges was evaluated in the anaesthetised guinea-pig using radioactive tracers, in parallel to the effects on cell counts in the broncho-alveolar lavage fluid, blood tumour necrosis factor (TNF-alpha) content, secretion of phospholipase A2 and synthesis of leukotriene C4 by alveolar macrophages. Intravenous injections of 0.1 or 1 mg/kg endotoxin induced lung leucocyte sequestration but only the higher dose induced an increase in albumin microvascular exchanges and the infiltration of leucocytes towards the airway lumen. Leukotriene B4, a potential mediator of the 5-lipoxygenase-dependent endotoxin effects, induced a rapid and transient lung leucocyte sequestration and leucopenia associated with a more progressive increase in microvascular exchanges. The 5-lipoxygenase inhibitor, BW B70C, injected i.p. (30 mg/kg) prevented leukotriene C4 synthesis by alveolar macrophages and reduced leucocyte migration to the airways lumen as well as albumin microvascular leakage but did not affect the endotoxin-induced increase in the blood level of TNF-alpha and of secreted phospholipase A2. However, BW B70C failed to modify vascular leucocyte margination induced by 1 mg/kg endotoxin, suggesting that, apart from a role of 5-lipoxygenase, alternative pathways operate in response to endotoxin in guinea-pig.

Topics: Animals; Blood Proteins; Bronchoalveolar Lavage Fluid; Bronchopulmonary Sequestration; Cell Count; Cell Separation; Dose-Response Relationship, Drug; Drug Interactions; Escherichia coli; Guinea Pigs; Hydroxylamines; Hydroxyurea; Injections, Intravenous; Isotope Labeling; Leukocytes; Leukopenia; Leukotriene B4; Leukotriene C4; Lipopolysaccharides; Lipoxygenase Inhibitors; Lung; Macrophage Activation; Macrophages, Alveolar; Methylurea Compounds; Phospholipases A; Phospholipases A2; Radioimmunoassay; Serum Albumin; Tumor Necrosis Factor-alpha

A series of (hydroxyphenyl)pyrazoles was designed by molecular modeling comparison with the LTB4 structure and prepared for evaluation as LTB4 receptor antagonists, culminating in 4-ethyl-5-[[6-methyl-6-(1H-tetrazol-5-yl)heptyl]oxy]-2-(1H-pyrazol -3- yl)phenol (2). Using an assay for inhibition of specific [3H]LTB4 binding to human PMN, it was found that the pyrazole ring could be methylated at N(1) with little loss of activity while methylation at N(2) reduced activity significantly. The structure-activity relationship of the terminal acid group was investigated. Good activity was found with o- and m-phenylalkanoic acids, chromane carboxylic acid, and tetrazole groups. The best in vitro activity was realized with the pyrazole nitrogen unsubstituted and with a six-carbon chain linking the phenyl ether oxygen to the tetrazole group. Compound 2, having an IC50 of 6.4 +/- 0.8 nM in the binding assay, was selected for further preclinical evaluation.

Topics: Animals; Cell Aggregation; Cells, Cultured; Humans; Leukopenia; Leukotriene B4; Models, Molecular; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Pyrazoles; Rabbits; Receptors, Leukotriene B4; Structure-Activity Relationship

SM-15178, a new hydroxyacetophenone derivative, was evaluated to determine its antiinflammatory activity and antagonistic activity against leukotriene B4 (LTB4). SM-15178 inhibited [3H]LTB4 binding to its receptors on human neutrophils (IC50 = 0.30 microM). It inhibited LTB4-induced chemotaxis of human neutrophils (IC50 = 0.72 microM) with little inhibitory effect against C5a or FMLP-induced chemotaxis at concentrations up to 30 microM. The compound alone did not cause human neutrophil chemotaxis at concentrations up to 10 microM. LTB4-induced chemotaxis of mouse and rat neutrophils and guinea pig eosinophils was also inhibited by the compound, with IC50 values of 0.55, 0.52, and 0.58 microM, respectively. In an in vivo study, SM-15178, given orally, significantly prevented LTB4-induced transient leukopenia. It also suppressed LTB4-induced bronchoconstriction in the guinea pig almost completely when given orally at a dose of 40 mg/kg. Furthermore, orally given SM-15178 suppressed arachidonic acid-induced neutrophil infiltration in mouse ears and Arthus reaction-induced paw edema in the mouse in a dose-dependent manner. These results suggest that SM-15178 is a selective and orally active LTB4 antagonist and that it might be effective for the treatment of some types of inflammatory diseases.

Topics: Acetophenones; Administration, Oral; Animals; Anti-Inflammatory Agents, Non-Steroidal; Chemotaxis, Leukocyte; Dose-Response Relationship, Drug; Guinea Pigs; Humans; Injections, Intravenous; Leukopenia; Leukotriene B4; Male; Mice; Mice, Inbred BALB C; Neutrophils; Rats; Rats, Sprague-Dawley

Exposure of rats to ozone (O3) produces an increase in airway permeability and a concomitant influx of polymorphonuclear leukocytes in the lung. These observations raise the possibility that the inflammatory cells play a role in the cellular injury and increased airway permeability after O3 exposure. This study was therefore designed to determine if the inflammatory cells or their products are essential for the O3 effect. In a series of experiments, rats were rendered leukopenic with cyclophosphamide, treated with leukotriene B4 (LTB4), or with the inhibitors of lipoxygenase or cyclooxygenase products of arachidonic acid, followed by exposure to O3. A 2-h exposure to 0.8 ppm O3 caused a significant increase in the flux of proteins and albumin in bronchoalveolar lavage (BAL) and elevated the transport of 99mTc-diethylenetriaminepentaacetate (99mTc-DTPA) from trachea to blood. The treatment with cyclophosphamide caused a significant reduction in the circulating and pulmonary leukocytes and prevented an increase in tracheal mucosal permeability to 99mTc-DTPA and the protein and albumin flux in BAL. While the intratracheal instillation of LTB4 did not affect the permeability, tracheal permeability and albumin levels in BAL in rats treated with LTD4 antagonist FPL 55712 and exposed to O3 were lower than in the untreated O3-exposed rats. Pretreatment with indomethacin also prevented the O3 effects, as reflected by the decreased protein and albumin flux in BAL and 99mTc-DTPA transport from trachea to blood. These data show a reduction in the effect of O3 by agents that affect leukocytes or their products. The results support a mechanism of increased permeability that is dependent upon inflammatory cells and their products.

Topics: Albumins; Animals; Blood Proteins; Bronchoalveolar Lavage Fluid; Chromones; Cyclophosphamide; Indomethacin; Leukocyte Count; Leukopenia; Leukotriene B4; Male; Ozone; Permeability; Pulmonary Alveoli; Rats; Rats, Inbred F344; Technetium Tc 99m Pentetate; Trachea

LY223982, (E)-5-(3-carboxybenzoyl)-2-((6-(4-methoxyphenyl)-5- hexenyl)oxy)benzenepropanoic acid, is a newly discovered potent inhibitor of specific binding of leukotriene B4 (LTB4) to its receptor on human neutrophils. This study demonstrated that the compound is also a specific antagonist of LTB4-induced neutrophil activation under both in vitro and in vivo conditions. LY223982 was found to be 189-fold more effective in displacing [3H]LTB4 than 35S-N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) from their corresponding receptors on human neutrophils. The concentration inhibiting 50% of response (IC50) for displacement of [3H]LTB4 (13.2 nM) was only 6.8-fold higher than the value for nonradioactive LTB4. The compound inhibited the aggregation of guinea pig neutrophils caused by LTB4 more strongly than FMLP or platelet-activating factor. The IC50 for inhibition of LTB4-induced responses (74 nM) was 93- and > 135-fold lower than the IC50 for inhibition of the corresponding FMLP and platelet-activating factor-induced effects. LY223982 was also a potent antagonist of the aggregation of human neutrophils by LTB4 (IC50, 100 nM). Chemotaxis of human neutrophils induced by LTB4 was only modestly inhibited by the compound (IC50 = 6 microM) but it had even less effect on cell movement caused by FMLP. LY223982 inhibited transient leukopenia induced in rabbits with LTB4 (ED50, 3 mg/kg) but not with FMLP. It had no agonist activity in any of the test systems. In summary, the results indicate that LY223982 is a potent specific antagonist of LTB4-induced neutrophil activation.

Topics: Animals; Benzophenones; Cell Aggregation; Chemotaxis, Leukocyte; Female; Guinea Pigs; Humans; Leukopenia; Leukotriene B4; Male; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Rabbits

Endotoxin plays an important role in the pathogenesis of septicaemia by activation of cellular and plasmatic systems. This study was performed to investigate the effects of infusion of endotoxin in rabbits by measuring the activation of cellular and plasma systems. Endotoxin was infused at a rate of 1 mg/kg body wt for 10 min, which caused death of all rabbits within 72 h. Endotoxin induced early leukopenia and thrombopenia, increased plasma levels of beta-glucuronidase and leukotriene B4 (LTB4), and decreased complement total hemolytic activity (CH50) and tissue plasminogen activator (t-PA) activity. These observations correlate with the cellular and plasma changes that have been documented in severely ill endotoxemic patients. Therefore, we conclude that this endotoxin model in rabbits is a valuable tool for investigation of pathophysiology and treatment of endotoxic shock.

Topics: Animals; Complement System Proteins; Endotoxins; Glucuronidase; Leukopenia; Leukotriene B4; Lipopolysaccharides; Rabbits; Shock, Septic; Thrombocytopenia; Tissue Plasminogen Activator

A method to quantify leukotriene B4-(LTB4)-induced changes in peripheral granulocyte counts in the rabbit is described. Rabbits were surgically prepared with vascular access ports cannulating the right external jugular vein. This preparation made possible rapid, accurate, and repeated sampling of venous blood. Intravenous infusion of LTB4 (0.5-2 micrograms/mL) into the left marginal ear vein was found reproducibly to cause an initial, rapid (1-5 min) leukopenia (64%-100% reduction) followed by an extended (20-30 min) leukocytosis (121%-178% increase). Saline infusion for 30 min resulted in no changes in peripheral granulocyte number. The method described was sensitive and reproducible enough to allow evaluation of the LTB4 receptor antagonist, LY223982 (10 mg/kg, i.v.), which was shown to block both the leukopenia and the leukocytosis induced by LTB4 infusion.

Topics: Animals; Benzophenones; Catheterization; Female; Granulocytes; Leukocyte Count; Leukopenia; Leukotriene B4; Neutrophils; Rabbits

Extracorporeal membrane oxygenation (ECMO) was performed in 27 mongrel adult dogs employing a V-V bypass between the proximal region of the inferior vena cava, used as a blood outlet, and the external jugular vein, used as a blood inlet, in order to investigate the decrease in leukocytes and the change in leukotriene B4 (LTB4) caused by ECMO. AA-861, an inhibitor of LTB4 activity, was given to 10 of the animals (AA-861 group), and the results obtained from this group were compared with those of a control group not given AA-861. Pulmonary arterial blood and pulmonary venous blood, for which femoral arterial blood was substituted, were sampled before ECMO, 5, 15, 30, 60, and 90 minutes after the beginning of ECMO, and 30 and 60 minutes after the termination of ECMO; measurements of various parameters, including WBC count, were carried out. The WBC count was seen to markedly drop immediately after the beginning of ECMO in both the control and AA-861 groups, and was significantly decreased in the femoral arterial blood than in the pulmonary arterial blood in the early phase of ECMO in both groups, revealing that the number of leukocytes differed between regions proximal and distal to the vital lung. This leukosequestration in the vital lung, pulmonary leukocyte sequestration in other words, was markedly inhibited in the AA-861 group than in the control group. Provided that leukosequestration is a phenomenon of leukocyte entrapment in the vital lung, a rate of entrapment could be calculated in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Dogs; Extracorporeal Membrane Oxygenation; Leukocyte Count; Leukocytes; Leukopenia; Leukotriene B4

1. Intravenous administration of ovalbumin (1 mg kg-1) to guinea-pigs that had previously been injected with 3.5 x 10(9) platelets from actively sensitized animals induced an approximately 40% decrease in the number of circulating leucocytes 30-60 min later, whereas the number of platelets was not affected. 2. In contrast, there was no change in the leucocyte number following antigen challenge of guinea-pigs that had received platelets from non-immunised animals. 3. This platelet-dependent leucopenia was inhibited by prior treatment of the recipient animal with cetirizine (10-30 mg kg-1, i.v.). Terfenadine (50 mg kg-1, p.o.) and mepyramine (2 mg kg-1, i.v.) were completely inactive in this respect. All doses of anti-histamines were used at concentrations which completely inhibited the bronchoconstriction to an i.v. injection of 5 micrograms kg-1 of histamine. 4. The site of action of cetirizine is most likely to be the platelet as leucopenia induced by the neutrophil agonists leukotriene B4 (LTB4) (30 ng kg-1) and platelet activating factor (PAF) (30 ng kg-1) were not modified by cetirizine treatment. 5. In these experiments, we failed to support a role for lipoxygenase products as mediators of the platelet-dependent leucopenia, as the selective lipoxygenase inhibitor BWA4C (50 mg kg-1, p.o.) was ineffective. 6. Our present results confirm and extend previous data demonstrating that antigen stimulated platelets can induce leucopenia in non-immunised animals and this can be inhibited by the anti-allergic agent, cetirizine, by an action which is probably unrelated to its anti-histamine properties. The precise nature of the platelet derived factor(s) and their target of action remains to be determined.

Topics: Animals; Benzeneacetamides; Benzhydryl Compounds; Blood Platelets; Cetirizine; Female; Guinea Pigs; Histamine H1 Antagonists; Hydroxamic Acids; Hydroxyzine; In Vitro Techniques; Leukocyte Count; Leukopenia; Leukotriene B4; Lipoxygenase Inhibitors; Male; Platelet Activating Factor; Pyrilamine; Terfenadine

Injection of leukotriene (LT)B4 (0.1-3 micrograms/kg i.v.) in normal anesthetized dogs produced dose-related leukopenia that was accompanied by arterial hypotension and tachycardia at higher tested doses. LTD4 (0.1-3 micrograms/kg i.v.), in contrast, increased arterial blood pressure, lowered cardiac rate and produced little change in arterial blood leukocyte count. Continuous infusion of LY255283 [(1-(5-ethyl-2-hydroxy-4-(6-methyl-6-(1H-tetrazol-5-yl)- heptyloxy)phenyl)ethanone] (0.33 mg/kg/min i.v.), a selective LTB4 receptor antagonist, resulted in near complete inhibition of leukopenic, hypotensive and tachycardic responses to LTB4 (3 micrograms/kg i.v.) challenge over a 6-hr test period. Persistent antagonism of canine LTB4 receptors was associated with high circulating levels of LY255283 that were bound extensively to plasma proteins. In subsequent experiments, myocardial infarct size was measured following 1 hr of occlusion of the circumflex coronary artery and 5 hr of reperfusion in control dogs infused with vehicle, and in dogs receiving LY255283 (0.33 mg/kg/min i.v.). Drug and vehicle were infused continuously beginning 15 min before coronary artery occlusion. LY255283 treatment essentially did not alter base-line cardiovascular parameters or myocardial oxygen demand when alterations were compared to time-related changes observed in control dogs. LY255283 infusion also did not alter the degree of myocardial ischemia or the intensity and duration of cardiac arrhythmias associated with coronary artery occlusion and reperfusion. Resultant infarct sizes were 43 +/- 5% of the left ventricle placed at risk in control dogs and 32 +/- 5% in dogs given LY255283; this difference was not statistically significant. The extent of left ventricle placed at risk was similar between groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Azoles; Blood Pressure; Coronary Circulation; Dogs; Heart Rate; Leukopenia; Leukotriene B4; Male; Myocardial Infarction; Neutrophils; Receptors, Immunologic; Receptors, Leukotriene B4; SRS-A; Tetrazoles

Lung injury following intravenous oleic acid is characterized by pulmonary edema, leukopenia and hypoxemia. Because leukotrienes can increase permeability and cause leukocyte adherence, we evaluated their potential role in oleic acid-induced lung injury in the anesthetized rat using a selective LTD4/E4 antagonist, LY171883. 99mTc-albumin and 99mTc-red blood cells (99mTc-RBC) were used to measure changes in the pulmonary permeability index and intravascular space by non-invasive scintigraphy. Intravenous oleic acid (0.06 ml/kg) increased the pulmonary permeability index 11 (P less than 0.01) and 5.8 fold (P less than 0.01) at 5 and 50 min after its injection compared to baseline, but had no effect on mean pulmonary arterial pressure or pulmonary distribution of 99mTc-RBC. Oleic acid also induced arterial hypoxemia, and increased bronchoalveolar lavage-fluid levels of immunoreactive (i) leukotriene LTC4 from 0.40 +/- 0.14 ng/ml to 2.27 +/- 0.55 ng/ml (mean +/- S.E.M., n = 4, P less than 0.05) and iLTB4 (from 0.42 +/- 0.05 ng/ml to 1.91 +/- 0.63 ng/ml, n = 5-7, P less than 0.01). LY171883 attenuated the elevated permeability by 24% and 68% at 5 (P less than 0.05) and 50 min (P less than 0.01), but did not alter the hypoxemia. These results support the hypothesis that oleic acid elevates leukotriene levels which may increase pulmonary vascular permeability. Furthermore, they suggest that the prevention of elevated pulmonary vascular permeability and edema may be necessary, but are clearly not sufficient to prevent arterial hypoxemia following oleic acid injury in the rat.

Topics: Acetophenones; Animals; Capillary Permeability; Dimercaprol; Leukopenia; Leukotriene B4; Leukotriene E4; Male; Oleic Acid; Oleic Acids; Oxygen; Pulmonary Edema; Rats; SRS-A; Tetrazoles

Serum from a patient with inactive systemic lupus erythematosus (SLE) and ibuprofen-induced transient neutropenia was used as a probe to define further the control of human polymorphonuclear leukocyte (PMN) exocytosis and superoxide (O2-) generation. Thirty-minute preincubation of normal PMNs with 10-50% v/v of this serum, followed by washing, produced a specific dose-related suppression of leukotriene B4 (LTB4)-elicited beta-glucuronidase and lysozyme release of up to 45% and 30% respectively. If cells were not washed, the inhibition increased to 60% and 40%. Superoxide production stimulated by LTB4 was unaffected. The serum had no effect on formyl-met-leu-phe (FMLP) or phorbol myristate acetate-stimulated O2- or exocytosis. O2- and beta-glucuronidase release elicited by zymosan-treated serum (ZTS) were both decreased by 15%, but there was no increased inhibition seen if cells were not washed, or if the time of preincubation was increased from 7 to 30 min. In contradistinction, the serum inhibition of LTB4 exocytosis did show time dependence. Serum obtained when the patient was not leukopenic and sera from 6 normal controls, 2 patients with inactive SLE, 1 patient with SLE and chronic leukopenia, and 2 controls taking ibuprofen did not influence any PMN function. The serum inhibition of ZTS-induced functions was qualitatively similar to that observed when PMNs were preincubated and desensitized with ZTS in vitro. Selective inhibition of LTB4 exocytosis was not seen when PMNs were desensitized with LTB4 in vitro. These observations indicate that LTB4-elicited O2- and exocytosis can be independently and specifically regulated. The cellular site at which this serum factor acts is not clear, but the current studies strongly suggest that this inhibition is not due to in vitro deactivation by LTB4 activity.

Topics: Exocytosis; Glucuronidase; Humans; Ibuprofen; Leukopenia; Leukotriene B4; Lupus Erythematosus, Systemic; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Stimulation, Chemical; Superoxides; Zymosan