Endotoxin-activated Serum Research Articles

Carbamazepine Affects Neutrophil Function through an Action on Peripheral Benzodiazepine Receptors

The aims of this study were to assess the possible role of peripheral benzodiazepine receptors (pBZrs)1 in mediating the in vitro effects of carbamazepine (CBZ) on some neutrophil functions in healthy volunteers and to investigate neutrophil function and pBZr expression in patients with epilepsia on CBZ monotherapy for at least 1 year. In vitro CBZ (42–168 μM) concentration-dependently inhibited chemotaxis induced by N-formyl-methionyl-leucyl-phenylalanine (FMLP) or lipopolysaccharide (LPS)-activated human serum. CBZ did not affect random migration, phagocytosis index, phagocytosis frequency, NBT reduction frequency, C. albicans lethality index and resting superoxide production. The pBZr antagonist PK 11195 (1 μM, per se ineffective) reversed the inhibitory effect of CBZ on chemotaxis induced by endotoxin-activated serum or FMLP. The pBZr agonist Ro 5–4864 (10–100 μM) mimicked the effect of CBZ on chemotaxis induced by endotoxin-activated serum or FMLP and had no effect on the other parameters. Neutrophils from epileptic patients on chronic CBZ monotherapy had impaired FMLP- and serum-induced chemotaxis and enhanced expression of pBZrs on neutrophils. These data strongly suggest an involvement of pBZrs in mediating the in vitro effects of CBZ on chemotaxis; furthermore, impairment of the same neutrophil function parameters and overexpression of pBZrs in patients are consistent with the hypothesis of an in vivo interaction of CBZ with pBZrs.

DEFECTIVE NEUTROPHIL (PMN) RESPONSE TO SALMONELLA (SAL) CHEMOTACTIC FACTORS(CF) in SAL GASTROENTERITIS (GE) 709

SAL infections are serious problems and especially dangerous among immuno compromised. We studied the role of PMN chemotaxis in 8 immuno competent infants between day 3 & 5 & 4 of them again after therapy along with same # of controls utilizing Boyden's technique. PMN rich WBC's(1.0×104) from the patients deposited onto a 3u filter and placed in the upper chambers were tested against 1) CF from pt's own SAL, 2) standard E. coli factors (ECF) 3) Endotoxin activated serum (EAS) and 4) Hank's solution (HS) placed in lower chambers in simultaneous expts performed in dulicate. After 3 hours' incubation and staining, migrated cells to lower surface were counted. The ratio of migrated to total was termed as chemotactic Index (C1) in first 3 expts & random migration (RM) in the fourth. Mean(ISD) of the duplicate, taken as final CI/RM are presented(Table). Mean CIs with each CF were lower than control(P <005) and RM's were similar. Post Rx CIs improved significantly. We conclude a) Defect of PMN chemotaxis occurs in SAL infection transiently b) Defect is not specific to SAL CF c) The defect may contribute to SAL invasiveness/virulence and more dangerously so among immuno comprised.

Heterophil Function in Healthy Chickens and in Chickens with Experimentally Induced Staphylococcal Tenosynovitis

Heterophil function was evaluated in 16 healthy chickens and in 46 chickens with experimentally induced staphylococcal tenosynovitis. In paired blood samples, heterophils from chickens with tenosynovitis had a significant increase in adherence, chemotaxis, phagocytosis, and bacterial killing of Staphylococcus aureus compared to heterophils from healthy chickens. The percent adherence of heterophils to nylon fiber columns increased significantly from a 78.4% mean +/- 6.6% standard deviation to 87.6% +/- 3.2% after induction of staphylococcal tenosynovitis. Heterophil movement following in vitro exposure to saline or endotoxin was increased in chickens with tenosynovitis; 3 +/- 1 heterophils/0.25 mm2 to 10 +/- 6 heterophils/0.25 mm2 and 136 +/- 29 heterophils/0.25 mm2 to 340 +/- 74 heterophils/0.25 mm2, respectively. Endotoxin-activated serum was chemoattractive for heterophils from all chickens. Flow cytometry was used to define the heterophil population on light scatter histograms, evaluate individual cell phagocytosis of latex beads, and quantitate the number of beads phagocytosed per heterophil. When incubated with increased numbers of beads, only heterophils from chickens with tenosynovitis phagocytosed higher numbers of beads. At heterophil to bead ratios of 1:10, the percentage of heterophils that phagocytosed beads increased from baseline values of 37.8% +/- 9.0% to post-infection values of 67.3% +/- 7.5%. Using 1:20 heterophil to bead ratios, heterophil phagocytosis increased from 38.7% +/- 9.9% to post-infection values of 79.8% +/- 7.3%. Heterophils from all chickens were able to phagocytose and kill log phase staphylococcal bacteria. After phagocytosis, the heterophils from chickens with staphylococcal tenosynovitis rapidly decreased the number of viable bacterial colony forming-units per milliliter by approximately one log.(ABSTRACT TRUNCATED AT 250 WORDS)

Isolation of blood granulocytes from golden hamsters for the study of chemotaxis

Since the number of granulocytes is low in the circulating pool of adult golden hamsters, intraperitoneal injection of oyster glycogen was used to mobilize white blood cells from the non-circulating pool. The percentage of blood granulocytes increased from 7 to 49%. Mobilized granulocytes, purified by a discontinuous Percoll gradient, showed adherence to polycarbonate filters only when coated by collagen. Random migration was high using the modified Boyden chamber technique. While endotoxin-activated serum caused chemotactic response, it was absent when the tripeptide N-formyl-methionyl-leucyl-phenylalanine was applied at concentrations of 10(-7) or 10(-8) M. The true chemotactic response corresponded to the proportion of migrated cells versus the total cell number adherent to both sides of the filter. It is concluded that the golden hamster is a good model for the study of granulocyte sequestration.

Preincubation with Intravenous Lipid Emulsion Reduces Chemotactic Motility of Neutrophils in Cord Blood

Neutrophils from cord blood of healthy term infants were isolated and incubated for 30 min with varying concentrations of intravenous lipid emulsion (ILE) solution (4, 8, 20 mg/ml). In vitro assay of chemotaxis was performed after incubation for 120 min with endotoxin-activated serum (EAS). Neutrophil random motility was unchanged after ILE incubation yet chemotactic factor (EAS)-stimulated motility was significantly reduced in a dose-related pattern.

The Role of Formylpeptide Receptors, C5a Receptors, and Cytosolic-Free Calcium in Neutrophil Priming

Polymorphonuclear leukocytes (PMNL) exposed to chemoattractants or cytokines change their functional capacity. The effect of endotoxin-activated serum as a priming agent on human PMNL was tested. Pretreatment of PMNL with endotoxin-activated serum increased their oxidative burst in response to formylpeptide (FMLP) (P less than .02) and C5a (P less than .05). Priming for membrane depolarization was observed in PMNL preincubated with either endotoxin-activated serum, low concentrations of purified C5a, or endotoxin but not with decomplemented plasma. Primed PMNL had an increased number of FMLP but not C5a receptors as compared with control PMNL. The "resting" cytosolic free calcium was increased in primed PMNL (P less than .02). Intracellular calcium buffering abolished the priming effect of endotoxin-activated serum. Thus, endotoxin-activated serum can prime cellular responsiveness for membrane depolarization and superoxide production in response to FMLP and to C5a. Priming may be due to an increased resting cytosolic-free calcium.

Improvement in alveolar macrophage migration after therapeutic whole lung lavage in pulmonary alveolar proteinosis.

Pulmonary alveolar proteinosis (PAP) is a rare disease characterized by the accumulation of lipoproteinaceous material in the alveolar space. Alveolar macrophages (AM) in this disease have been noted in the past to have abnormally decreased function. It is not known whether these abnormalities are reversible with treatment of the disease. In this study, the effect of therapeutic whole lung lavage (WLL) on AM migration in four patients with PAP was determined. Standard segmental bronchoalveolar lavage (BAL) was performed 1 day before and 6 days after WLL. AM recovered by BAL were assayed for migratory response to N-formyl-methionyl phenylalanine (FMP) and endotoxin-activated human serum (EAHS). As expected, migration toward both stimuli was decreased before WLL compared to a group of normal controls. After WLL, AM migration toward both stimuli was not only greater than before the WLL but also was greater than normal. In one patient, serial BAL over a period of 18 months showed a marked decline in AM migration coincident with a clinical relapse of the disease. Thus, the defect in AM function found in PAP is reversible with WLL but can recur if clinical remission is not sustained.

Depressed leukotriene B4 chemotactic response of neutrophils from localized juvenile periodontitis patients.

Previous studies have demonstrated that certain local juvenile periodontitis (LJP) patients possess neutrophils with an intrinsic chemotactic defect to the peptide chemoattractants f-met-leu-phe (FMLP) and C5a. In this investigation, the in vitro response of neutrophils to the chemotactic agent leukotriene B4 (LTB4) was examined. Those LJP patients who possessed defective chemotactic responses to FMLP and endotoxin activated serum (EAS) also had impaired chemotaxis to LTB4. Exogenous LTB4 failed to augment the impaired response to FMLP. Evidence for a global membrane defect in LJP neutrophils resulting in hyporesponsiveness to chemically diverse chemotactic agents is presented.

Shift in subfractions of rat alveolar macrophages in vivo during endotoxin-induced alveolitis.

To elucidate changes in alveolar macrophages that accompany sepsis-induced lung injury, this study analyzed the subfractions of alveolar macrophages (AM) recovered by lung lavage during the onset of endotoxin-induced acute neutrophilic alveolar inflammation in the rat model. Centrifugation on continuous self-generated density gradients of Percoll was used to fractionate AM into subpopulations between density limits 1.012 and 1.130. Two-thirds of AM recovered from pathogen-free control rats (group C) were in a fraction with a density range of 1.058-1.078 ["normal" density fraction, (ND)]. Only 6% were located in a very low density (VLD) fraction 1.037-1.048. Neutrophils accounted for less than 1% of recovered cells and usually were found in the fraction with density range of 1.079-1.130. By contrast, if rats underwent lung lavage 15 hours after the administration of endotoxin (group E), only 38% of macrophages were recovered in the "normal" density fraction, whereas 26% of the AM recovered were in the VLD fraction. This shift in the relative sizes of the density based subpopulations coincided with the onset of acute bronchoalveolar inflammation as indicated by the recovery of neutrophils by bronchoalveolar lavage (PMN = 7 X 10(4) in C, vs. 9.4 X 10(5) in E, p less than .001). The macrophages on the low density subfractions showed functional impairment: they were less viable in culture and migrated poorly in response to endotoxin-activated serum compared to macrophages in the "normal" density fraction from the endotoxin-treated animals. The rapid emergence of the low density population after endotoxin could represent an influx of new cells, but more likely indicates that injury to or previous activation of resident macrophages has caused their density to decrease. We speculate that the emergence of a population of AM in airspaces with low density and impaired function could weaken pulmonary host defence following endotoxemia.

NEUTROPHIL (PMN) MIGRATION IN RESPONSE TO CHEMOTACTIC FACTORS GENERATED BY THE INVADING PATHOGENS IN ACUTE BACTERIAL ENTERITIS (ABE)

The role of chemotactic and random migration(RM) in ABE is not known. They were determined in 9 pts with ABE due to shigella or salmonella between days 3&5 of admission and 9 controls. One day prior to study each isolate was grown in medium 199 to generate specific chemotactic factors (SCF). Leukocytes were harvested from heparinized blood and 1.0×106PMNS were deposited on to 3 u millipore filter which was placed in the Boyden's chamber. The upper compartment of the chamber was filled with Hank's solution (HS) and the lower with SCF 100uL/mL of HS. Three simultaneous chambers were also set up with endotoxin activated serum (EAS), E. coli generated factors (ECF) and HS alone in the lower compartments. After 3hr's incubation filters were stained and cells which migrated to lower surface were counted. A ratio of migrated to total cells with SCE, EAS & ECF was termed as chemotactic index(CI) and RM with HS alone. In 4 pts studies were repeated after therapy. Mean (±1SD) values are presented (table): Mean CIs of patients with all the 3 factors were significantly lower than those of controls and significantly increased after therapy. RM was similar. Supression PMN chemotaxis may be one step in the pathogenesis of ABE. Counteracting it with drug/agents may improve the outcome.

ｅｎｄｏｔｏｘｉｎ活性化血清による好中球ａｃｅｔｙｌｔｒａｎｓｆｅｒａｓｅ活性の増強

Endotoxin-activated serum, but not endotoxin or serum alone, had a stimulatory effect on acetyltransferase activity (AT) of neutrophils, which is a key enzyme in biosynthesis of platelet-activating factor. The AT stimulatory factor in endotoxin-activated serum was heat-stable, undialysable, and its activity was linear to that of endotoxin between 0.05-5μg/ml. Zymosan-activated serum, which is known to contain a substantial amount of C5a, induced superoxide anion production and degranulation of neutrophils, but did not elevate the AT level of neutrophils. On the other hand, endotoxin-activated serum did not induce superoxide anion production or degranulation. These findings provided evidence against the role of C5a in induction of AT. Sephadex G-200 gel chromatograhy and protein A affinity chromatograhy suggest that modified IgG molecules play a role in elevating the AT level of neutrophils.

Neutrophil function in localized juvenile periodontitis. Phagocytosis, superoxide production and specific granule release.

Patients with localized juvenile periodontitis (LJP) exhibit defective neutrophil functions to a variety of environmental and host stimuli. It is not clear, however, how many of the measurable functions are defective and whether individual patients exhibit single or multiple dysfunctions. The purpose of this study was to evaluate chemotaxis, phagocytosis, specific granule release and superoxide production in a group of 23 previously unreported LJP patients. Our results indicate that all 23 of these LJP patients exhibited chemotaxis depression to N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) and endotoxin-activated serum (EAS). Smaller groups from the 23 chemotactically defective LJP group were used to test other function due to inability to obtain sufficient quantities of blood. Fourteen of 14 LJP patients tested exhibited defective phagocytosis. Ten LJP patients were evaluated for specific granule release, and 14 LJP patients were evaluated for superoxide production. Both granule release and superoxide production were found to be normal in chemotactically defective LJP patients. Since both defective and normal responses noted in the same neutrophil populations are mediated by the same receptor, it is hypothesized that the cellular defect lies in a post receptor pathway.

Changes in the structural and functional properties of human eosinophils during experimental hookworm infection.

Normal volunteers were infected with hookworm larvae Necator americanus. Peripheral blood counts showed a mean of 524 +/- 29 eosinophils/mm3 of blood before infection and a mean of 3,008 +/- 456 eosinophils/mm3 of blood during infection (P less than .01). Absolute numbers of neutrophils did not change. Eosinophils and neutrophils from the infected period were compared with the noninfected state in each subject. The percentage of hypodense eosinophils increased from a mean of 34% +/- 13% to 80% +/- 7% during infection (P less than .05). Superoxide production of eosinophils increased from a mean of 56 +/- 9 to 97 +/- 12 nmol of O2-./10(6) cells per 60 min (P less than .05) during infection. Chemotaxis of eosinophils to Escherichia coli endotoxin-activated serum increased from a mean average distance migrated of 19 +/- 2 micron (P less than .05), whereas neutrophil responsiveness did not change. This is the first report of changes in eosinophil density and stimulation of eosinophil function in normal hosts experimentally infected with hookworm. The data indicate that hookworm infection preferentially increases eosinophil production and activity with little effect on neutrophils.

Human protein HC and its IgA complex are inhibitors of neutrophil chemotaxis.

Protein HC, a heterogeneously charged low molecular weight glycoprotein, and its IgA complex were isolated from human plasma and urine. Plasma from individuals with monoclonal IgA populations was used as starting material for the isolation of the protein HC-IgA complex to obtain homogeneous complex populations. Neither low molecular weight protein HC nor its IgA complex in the concentrations 30 and 600 mg/liter influenced the random migration of normal human neutrophils. The chemotactic response of neutrophils to endotoxin-activated serum was, however, attenuated in a dose-dependent way by both low molecular weight protein HC and protein HC-IgA complex. Concentrations of protein HC and its IgA complex producing significant inhibition of the chemotactic response were found to occur in plasma from healthy and diseased individuals as well as in synovial fluid from patients with rheumatoid arthritis. These results suggest that protein HC and its IgA complex play physiological roles in the regulation of the inflammatory response.

A novel leukocyte differentiation antigen: two monoclonal antibodies TM2 and TM3 define a 120-kd molecule present on neutrophils, monocytes, platelets, and activated lymphoblasts

We produced two hybridomas by fusion of mouse myeloma cells with splenocytes from a mouse immunized with the THP-1 human monocytoid leukemia cell line. Two cloned hybridoma cell lines, designated as TM2 and TM3, were obtained. They secreted antibodies against a unique cell surface antigen expressed on all normal peripheral blood monocytes, neutrophilic granulocytes, platelets, and mitogen-induced lymphoblasts, some cells from patients with immature-type lymphoid leukemias. However, the antibodies reacted neither with large numbers of peripheral blood lymphocytes nor with red cells. Cross-blocking studies showed that these monoclonal antibodies recognized the same or a nearly positioned antigen epitope. Immunoprecipitation of THP-1 cell extract with TM2 or TM3 under reducing and nonreducing conditions yielded a specific band of mol wt equal to 120,000 daltons. This determinant appeared to be involved in granulocyte chemotaxis, since neutrophilic granulocytes exposed to TM2 or TM3 showed a significant decrease in chemotaxis toward endotoxin-activated serum. These two monoclonal antibodies did not affect O2- release or luminol-dependent chemiluminescence of neutrophils. Moreover, they did not alter platelet aggregation induced by thrombin. TM2 and TM3 will provide a new reagent in defining the linkage between lymphoid and myeloid differentiation and intermyeloid development.

A Novel Leukocyte Differentiation Antigen: Two Monoclonal Antibodies TM2 and TM3 Define a 120-kd Molecule Present on Neutrophils, Monocytes, Platelets, and Activated Lymphoblasts

We produced two hybridomas by fusion of mouse myeloma cells with splenocytes from a mouse immunized with the THP-1 human monocytoid leukemia cell line. Two cloned hybridoma cell lines, designated as TM2 and TM3, were obtained. They secreted antibodies against a unique cell surface antigen expressed on all normal peripheral blood monocytes, neutrophilic granulocytes, platelets, and mitogen-induced lymphoblasts, some cells from patients with immature-type lymphoid leukemias. However, the antibodies reacted neither with large numbers of peripheral blood lymphocytes nor with red cells. Cross-blocking studies showed that these monoclonal antibodies recognized the same or a nearly positioned antigen epitope. Immunoprecipitation of THP-1 cell extract with TM2 or TM3 under reducing and nonreducing conditions yielded a specific band of mol wt equal to 120,000 daltons. This determinant appeared to be involved in granulocyte chemotaxis, since neutrophilic granulocytes exposed to TM2 or TM3 showed a significant decrease in chemotaxis toward endotoxin-activated serum. These two monoclonal antibodies did not affect O2- release or luminol-dependent chemiluminescence of neutrophils. Moreover, they did not alter platelet aggregation induced by thrombin. TM2 and TM3 will provide a new reagent in defining the linkage between lymphoid and myeloid differentiation and intermyeloid development.

Rapid Method for Isolation of Normal Human Peripheral Blood Eosinophils on Discontinuous Percoll Gradients and Comparison With Neutrophils

Previous studies on human eosinophils often have used cells from patients with hypereosinophilia syndrome or parasitosis owing to the difficulty in isolating pure populations of eosinophils from normal individuals. In the present study, human eosinophils were isolated with a purity of 97%, with 70% recovery from normal individuals with blood eosinophil counts of less than 3%. Human eosinophils are denser than neutrophils, but the range of densities of the two cell types overlap, making purification of eosinophils by density-gradient centrifugation difficult. However, if neutrophils were exposed to the chemotactic peptide (f-Met-Leu-Phe), which did not stimulate eosinophils, the neutrophils' density decreased, shifting them away from the density of eosinophils. Whole normal blood anticoagulated with EDTA was incubated at 37 degrees C for 15 minutes with 10(-6) mol/L f-Met-Leu-Phe and then layered over a discontinuous Percoll gradient (65% and 75% in diluted phosphate-buffered saline) and centrifuged at 400 g for 25 minutes at 22 degrees C. The cell layer between the 65% and 75% Percoll was collected and washed, and hypotonic lysis was used to remove erythrocytes. This cell layer contained 97.3 +/- 0.7% eosinophils (N = 8) with a yield of 4.9 X 10(4) eosinophils per milliliter of whole blood, or 70% of the total eosinophil count. The isolated eosinophils were in a quiescent state but responded to Escherichia coli endotoxin-activated serum with shape change and chemotaxis, membrane depolarization, and reduced nitroblue tetrazolium (96.0 +/- 1.0%), when stimulated with phorbol myristate acetate. In phagocytic assays, 89.3 +/- 1.3% of the eosinophils ingested Candida albicans v 96.0% +/- 1.0% of neutrophils. In contrast, the eosinophils did not respond chemotactically, alter membrane potential, or reduce nitroblue tetrazolium when treated with f-Met-Leu-Phe, and studies with f-Met-Leu-[3H]Phe showed that normal eosinophils lacked expression of receptors for f-Met-Leu-Phe. In control studies, normal eosinophils that were not exposed to f-Met-Leu-Phe during purification also failed to respond to f-Met-Leu-Phe, indicating intrinsic differences between normal eosinophils and neutrophils. Thus, exposure of whole blood to f-Met-Leu-Phe, followed by separation on Percoll is a simple method for rapid isolation of normal human eosinophils.

Functions of polymorphonuclear (PMN) leukocytes of patients with lymphoproliferative diseases

PMN leukocytes from untreated patients with multiple myeloma (MM), Hodgkin's disease (HD) and non-Hodgkin lymphoma (NHL) were studied in vitro for their phagocytic and chemotactic function. Alkaline phosphatase score and the reduction of nitroblue tetrazolium (NBT) in these leukocytes were also determined. Most of the functions of PMN leukocytes from untreated patients with MM were impaired, compared to control leukocytes, while those from patients with HD and NHL were impaired only in their chemotactic response to casein and endotoxin-activated serum (EAS).

Chemotactic and candidacidal responses of rabbit alveolar macrophages during postnatal development and the modulating roles of surfactant in these responses.

Chemotactic responses of alveolar macrophages from 1-, 7-, and 28-day-old rabbits to various concentrations of endotoxin-activated serum and n-formyl-methionyl-phenylalanine were tested utilizing both blind well and agarose plate assay systems. A dramatic increase in both the chemotactic response and responsiveness to various concentrations of chemoattractant was observed during postnatal maturation. The pattern of result was similar with both methods of assay. An age-related increase was also found to occur in the candidacidal activity of alveolar macrophages in contrast to their phagocytic uptake, which showed no age-related increases. Furthermore, the decreased function of macrophages from newborn animals correlated with a morphologically and biochemically less mature cell population which contained large amounts of phagocytosed surfactant-related material. Moreover, pretreatment of macrophages from 7- and 28-day-old animals with vesicles of surfactant-related material resulted in decreases in both chemotactic and candidacidal activity, with a paradoxical increase in their phagocytic activity. The resulting activities were similar to those of macrophages from 1-day-old animals treated with buffer alone. These data suggest that there is an age-related increase in the chemotactic and candidacidal activity of alveolar macrophages during maturation and that the decreased activity of macrophages from newborn animals is related in part to the large amount of surfactant-related material present at that time.

Benoxaprofen: a pro-oxidant anti-inflammatory drug?

Benoxaprofen inhibited the random motility and migration to the leucoattractants endotoxin-activated serum (EAS) and f-met-leu-phe of human polymorphonuclear leucocytes (PMNL) in vitro. Inhibition of random and leucoattractant-induced migration was observed at drug concentrations of greater than 1 X 10(-6) M and 1 X 10(-5) M respectively. Benoxaprofen per se was not leucotactic but was pro-oxidative in that it stimulated PMNL hexose-monophosphate shunt activity, chemiluminescence, myeloperoxidase-mediated iodination reactions and degranulation. The drug also mediated auto-oxidation of PMNL as measured by cellular auto-iodination. The relationship between benoxaprofen-mediated inhibition of PMNL migration and activation of oxidative metabolism was investigated using the anti-oxidants ascorbate and levamisole at concentrations of 10(-2) M and 10(-3) M respectively. These agents prevented the decreased motility and auto-oxidation of PMNL induced by 10(-4) M benoxaprofen. Benoxaprofen (10(-4) M) did not inhibit the migration of PMNL from 3 children with chronic granulomatous disease thus showing that intact PMNL oxidative metabolism is required for the induction of drug-mediated inhibition of cell motility. Ingestion of therapeutic doses of benoxaprofen for 7 days by normal adults gave serum drug concentrations greater than those required for detectable effects on PMNL functions in vitro (mean serum value 126 micrograms/ml). Co-incubation of normal PMNL with serum from individuals who had ingested the drug caused decreased cell migration and increased chemiluminescence. These results show that benoxaprofen inhibits PMNL migration as a consequence of pro-oxidant properties and despite its withdrawal may be the prototype of the pro-oxidative anti-inflammatory drug.