interleukin-8 and Peritonitis

Cytokines are pluripotent pleiotropic agents that have received widespread attention over the last few years. Not surprisingly, the have also been studied in the context of continuous ambulatory peritoneal dialysis. Cytokines play a central role in this treatment modality for uremic patients, because these inflammatory mediators act upon the biological dialysis membrane, i.e. the peritoneum, while simultaneously they participate in host defense mechanisms. This review describes which cytokines are present in dialysate, whether there is support for intraperitoneal release, and under which circumstances. If focuses particularly on the relationship between cytokines in peritoneal effluent and peritoneal permeability to macromolecules. In addition, the presence of prostanoids in dialysate and their role in the local regulation of peritoneal permeability are discussed, because cyclooxygenase products are tightly linked to cytokine networks.

Topics: Animals; Cell Membrane Permeability; Cytokines; Humans; Interleukin-1; Interleukin-8; Macrophages, Peritoneal; Peritoneal Dialysis, Continuous Ambulatory; Peritoneum; Peritonitis; Prostaglandins; Tumor Necrosis Factor-alpha; Uremia

This study was designed to evaluate differences in both the peritoneal and systemic immune response after laparoscopic and conventional surgical approaches.. Patients with a primary carcinoma were prospectively randomized to curative laparoscopic (n = 12) or conventional (n = 14) colon resection. The proinflammatory cytokines interleukin-6, interleukin-8, and tumor necrosis factor-alpha were measured in the peritoneal drain fluid and in the serum. C-reactive protein and leukocyte counts and the differences in leukocyte subpopulations and expression of human leukocyte antigen-DR on monocytes were measured perioperatively.. Significantly higher levels of proinflammatory cytokine were found in the peritoneal drain fluid than in the circulation after both procedures. Serum interleukin-6 and interleukin-8 levels were significantly lower 2 hours after laparoscopic surgery than with the conventional procedure. Postoperative cellular immune counts and human leukocyte antigen-DR expression normalized earlier after the laparoscopic approach.. The systemic proinflammatory concentrations after both surgical approaches represent only a small fragment of what is generated in the peritoneal drain fluid. Even if the immediate levels of proinflammatory cytokines in the serum are significantly lower in the laparoscopic group, the same cytokines locally produced showed no differences, which suggests that the two intra-abdominal approaches are equally traumatic. No differences in cellular response were observed between the groups.

Topics: Aged; Antibodies, Monoclonal; Ascitic Fluid; C-Reactive Protein; Carcinoma; CD4-CD8 Ratio; Colectomy; Colorectal Neoplasms; Cytokines; Female; HLA-DR Antigens; Humans; Interleukin-6; Interleukin-8; Laparoscopy; Male; Monocytes; Peritoneum; Peritonitis; Prospective Studies; Tumor Necrosis Factor-alpha

Continuous ambulatory peritoneal dialysis (CAPD) carries a risk of peritonitis which is accompanied by mild symptomatology. Culture of effluent has yielded organism in 50% of cases. Peritoneal phagocytes produce tumor necrosis factor-alpha and interleukin (IL)-1 in response to contact with bacteria, initiating an inflammatory cascade which leads to IL-6 and IL-8 secretion. Additonally, neutrophils undergo an increase in oxidative metabolism. We have evaluated the diagnostic accuracy of effluent measurements of TNF-alpha, IL-6, IL-8, and oxidative metabolism markers in these patients. Dialysate fluids (n = 65) were collected from non-infected patients and those presenting with acute peritonitis. Positive culture proved the diagnosis. Oxidative markers and nitric oxide were measured by chemiluminescence. Cytokines were measured by solid phase chemiluminescent immunometric assay (Immulite, DPC, USA). Receiver operating characteristic (ROC) curves were used to assess the diagnostic accuracy and the areas under curves were calculated for comparison. All effluent cytokines and oxidative markers were significantly higher in patients with peritonitis when compared to those without (p < 0.05). Significant correlations were evident between IL-6 and IL-8, lucigenin chemiluminescence and luminol chemiluminescence, lucigenin chemiluminescence and IL-6 or IL-8, and luminol chemiluminescence and IL-6 or IL-8. ROC curves showed that the ability of IL-6, IL-8, lucigenin chemiluminescence, and luminol chemiluminescence to differentiate CAPD patients with peritonitis from non-infected cases exceeds that of polymorphonuclear leukocyte count.

Topics: Adult; Aged; Ascitic Fluid; Cell Count; Cytokinins; Diagnosis, Differential; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Neutrophils; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; ROC Curve; Tumor Necrosis Factor-alpha

Topics: Ascites; Ascitic Fluid; Bacterial Infections; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Interleukin-8; Liver Cirrhosis; Luminescent Measurements; Neutrophils; Peritonitis

Efficacy and safety of panipenem/betamipron (PAPM/BP) in treatment of obstetric and gynecological infections, and change of interleukin-6 (IL-6) and interleukin-8 (IL-8) levels in blood, as markers of infection, were investigated. The results were as follows; 1) Clinical efficacy of PAPM/BP by drip infusion of 1-2 g/day for 3-14 days against 52 patients with intrauterine infection (n = 29), pelveoperitonitis (n = 19), and other infections were 14 "Excellent" in 14 cases, "Good" in 35 cases, and efficacy rate was 94.2% (49/52). Both efficacy rate analy by causative organisms and eradication rate were 35/37 (94.6%). No subjective or objective side effects and no abnormal labolatory findings were observed. 2) Changes of IL-6 (> 4 pg/ml) levels in serum, as an infection marker, were observed in 8 cases out of 14 cases (57.1%), and correlation between CRP and IL-6 in the treatment process was noticed. However, changes of serum IL-8 (> 12.5 pg/ml) were observed in only 2 cases of those 14 cases (14.3%), indicative that IL-8 has no significance as a marker of infection.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Bacterial Infections; beta-Alanine; Biomarkers; Drug Therapy, Combination; Female; Genital Diseases, Female; Humans; Interleukin-6; Interleukin-8; Middle Aged; Pelvic Inflammatory Disease; Peritonitis; Thienamycins; Uterine Diseases

A total of 105 patients participated in this study, including 10 with chronic glomerulonephritis with normal renal function (CGN patients), 36 uraemic patients (CRF patients), 19 continuous ambulatory peritoneal dialysis patients (CAPD) without peritonitis, three CAPD patients with peritonitis, 37 patients undergoing chronic haemodialysis (HD) divided into short-term HD, 15 patients; medium-term HD, 12 patients; and long-term HD, 10 patients. IL-8 and two other proinflammatory cytokines, IL-6 and TNF alpha were tested using a specific immunoassay. IL-8, IL-6, and TNF alpha serum levels were significantly increased in patients with chronic renal failure compared to their levels in normal individuals (P < 0.0001, P < 0.05 and P < 0.0001 respectively). The most pronounced increment in IL-8, IL-6 and TNF alpha serum levels was observed in CAPD patients (P < 0.0001). CAPD patients without peritonitis showed relatively low levels of IL-8 or IL-6 in peritoneal dialysate effluents (PDE), whereas PDE-TNF alpha were not detectable in almost all patients tested. Patients with peritonitis showed very high serum and PDE levels of IL-8, IL-6 and TNF alpha. The clinical recovery from peritonitis was characterized by a rapid fall in IL-8, IL-6 and TNF alpha in serum and dialysate. HD patients showed a significant increase in serum levels of IL-8 and also IL-6 and TNF alpha compared to normal individuals (P < 0.05, P < 0.05 and P < 0.01 respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Chronic Disease; Female; Glomerulonephritis; Humans; Interleukin-6; Interleukin-8; Kidney Diseases; Kidney Failure, Chronic; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Renal Dialysis; Renal Replacement Therapy; Tumor Necrosis Factor-alpha; Uremia

Interleukin-8 (IL-8) has been reported to be released by activated peritoneal macrophages (PMs) and a variety of other cell types, and it exhibits potent chemotactic activity for polymorphonuclear cells (PMNs). We have previously shown that IL-8 is detectable in the drain dialysate of uremic patients on continuous ambulatory peritoneal dialysis (CAPD) during peritonitis. The levels of IL-8 in infected drain dialysate caused by different microorganisms were variable. In this study, we evaluated the gene expression and release of IL-8 by PMs and PMNs during peritonitis caused by Staphylococcus aureus in uremic patients on CAPD. IL-8 levels were variable in the drain dialysate at the different episodes of peritonitis, even in the same patient. The IL-8 levels were highly correlated with PMN count in drain dialysate (r = 0.9919, p < 0.001). PMs and PMNs obtained from drain dialysate at the onset of peritonitis increased mRNA expression for IL-8 and the amount of IL-8 mRNA from drainage cells was also highly correlated with PMN count. In contrast, cells isolated from drain dialysate without peritonitis failed to express mRNA for IL-8. These data suggest that increased expression of IL-8 may be a feature of peritonitis. The levels of IL-8 during peritonitis were not only related to the etiological microorganism but also to other unknown factor(s).

Topics: Adult; Female; Gene Expression; Humans; Interleukin-8; Kidney Failure, Chronic; Macrophages, Peritoneal; Male; Middle Aged; Neutrophils; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; RNA, Messenger; Staphylococcal Infections; Uremia

Topics: Adult; Aged; Aged, 80 and over; Bacterial Infections; Biomarkers; DNA, Bacterial; Female; Fibrosis; Humans; Inflammation; Interleukin-6; Interleukin-8; Liver Cirrhosis; Male; Microbiota; Middle Aged; Peritonitis; RNA, Ribosomal, 16S

Alterations of the innate immunity contribute to the development of spontaneous bacterial peritonitis (SBP) in liver cirrhosis. Given its role in immune signaling, antimicrobial function, and macrophage differentiation, we hypothesized that genetic polymorphisms of TRAF6 modulate the risk of SBP. Thus, we determined theTRAF6 haplotype in 432 patients with cirrhosis and ascites using the haplotype-tagging single nucleotide polymorphisms rs331457 and rs5030419. In addition, peritoneal macrophages were immunomagnetically isolated and characterized. Overall, 122 (28%) patients had an episode of SBP. In the combined prospective-retrospective analysis the frequency of SBP differed between the four haplotypes (P = 0.014) and was the highest in 102 patients carrying the rs331457 but not the rs5030419 variant, when compared to other haplotypes (odds ratio 1.95 [1.22-3.12]) or to the wild-type (odds ratio 1.71 [1.04-2.82]). This association was confirmed in multivariate logistic regression (adjusted odds ratio 2.00 [1.24-3.22]) and in prospective sensitivity analysis (hazard ratio 2.09 [1.08-4.07]; P = 0.03). The risk haplotype was associated with lower concentrations of the immune activation marker soluble CD87 in ascitic fluid and with a decreased expression of IL-6 and CXCL8 in isolated peritoneal macrophages. In conclusion, genetic polymorphisms of TRAF6 are associated with decreased peritoneal immune activation and an increased risk of SBP.

Topics: Aged; Ascites; Female; Gene Expression; Haplotypes; Humans; Immunity, Mucosal; Interleukin-6; Interleukin-8; Intracellular Signaling Peptides and Proteins; Liver Cirrhosis; Logistic Models; Macrophages, Peritoneal; Male; Middle Aged; Odds Ratio; Peritoneum; Peritonitis; Polymorphism, Single Nucleotide; Primary Cell Culture; Prospective Studies; Receptors, Urokinase Plasminogen Activator; Retrospective Studies; Risk Factors; TNF Receptor-Associated Factor 6

Citral is an active compound of lemongrass oil which has been reported to have anti-inflammatory effects. In this study, we investigated the effects of citral on lipopolysaccharide (LPS)-induced inflammatory response in a rat model of peritonitis and human umbilical vein endothelial cells (HUVECs). LPS was intraperitoneally injected into rats to establish a peritonitis model. The HUVECs were treated with citral for 12 h before exposure to LPS. The levels of TNF-α and IL-8 were measured using ELISA. Western blotting was used to detect the expression of VCAM-1, ICAM-1, NF-κB, and PPAR-γ. The results showed that citral had a protective effect against LPS-induced peritonitis. Citral decreased the levels of WBCs and inflammatory cytokines TNF-α and IL-6. Citral also inhibited LPS-induced myeloperoxidase (MPO) activity in the peritoneal tissue. Treatment of HUVECs with citral significantly inhibited TNF-α and IL-8 expression induced by LPS. LPS-induced VCAM-1 and ICAM-1 expression were also suppressed by citral. Meanwhile, we found that citral inhibited LPS-induced NF-κB activation in HUVECs. Furthermore, we found that citral activated PPAR-γ and the anti-inflammatory effects of citral can be reversed by PPAR-γ antagonist GW9662. In conclusion, citral inhibits LPS-induced inflammatory response via activating PPAR-γ which attenuates NF-κB activation and inflammatory mediator production.

Topics: Acyclic Monoterpenes; Anilides; Animals; Anti-Inflammatory Agents; Cell Adhesion; Enzyme Activation; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Leukocyte Count; Lipopolysaccharides; Monoterpenes; Neutrophils; NF-kappa B; Peritonitis; Peroxidase; Plant Oils; PPAR gamma; Rats; Terpenes; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

The immune status was studied while surgical treatment of 74 patients, suffering extended peritonitis (EP). In 38 patients (group of comparison) a complex routine therapy was conducted, in 36 (the main group)--along with routine complex measures of treatment a local and systemic ozonotherapy, using ozonized perftoran (OP) and glutoxim (for immunocorrection), were applied. The patients severity state was estimated in accordance to the Mahnheim's peritoneal index. In all the patients while hospitalization there were revealed changes in T--and B--links of immunity, in phagocytosis and the cytokins profile, significant of which have depended upon the EP severity. The combined local and systemic application of OP and glutoxim in complex of conservative treatment is pathogenetically substantiated, it permits more rapidly to eliminate the immunity disorders and the cytokines balance, as well as inflammatory process in abdominal cavity and to improve the results of surgical treatment significantly.

Topics: Adolescent; Adult; Aged; B-Lymphocytes; Blood Substitutes; Case-Control Studies; Drug Therapy, Combination; Female; Fluorocarbons; Humans; Immunity, Innate; Immunologic Factors; Interleukin-4; Interleukin-8; Male; Middle Aged; Oligopeptides; Ozone; Peritonitis; Severity of Illness Index; T-Lymphocytes; Tumor Necrosis Factor-alpha

Allergy is characterized by eosinophilia and an increased susceptibility to microbial infection. Atopic dermatitis (AD) is typically associated with Staphylococcus aureus (SA) colonization. Some of the mechanisms by which SA and its exotoxins interact with eosinophils remain elusive. CD48, a glycosylphosphatidylinositol-anchored receptor belonging to the CD2 family, participates in mast cells-SA stimulating cross-talk, facilitates the formation of the mast cell/eosinophils effector unit and as expressed by eosinophils, mediates experimental asthma.. To investigate the role of CD48 expressed on human peripheral blood and mouse bone marrow-derived eosinophils (BMEos) in their interaction with heat-killed SA and its three exotoxins, Staphylococcal enterotoxin B (SEB), protein A (PtA) and peptidoglycan (PGN).. Eosinophils were obtained from human peripheral blood and BM of WT and CD48-/- mice. SA was heat killed and eosinophils-SA/exotoxins interactions were analyzed by confocal microscopy, adhesion and degranulation, cell viability, cytokine release and cell signalling. In addition, peritonitis was induced by SEB injection into CD48-/- and WT mice. CD48 expression was studied in AD patients' skin and as expressed on their leucocytes in the peripheral blood.. We provide evidence for the recognition and direct physical interaction between eosinophils and SA/exotoxins. Skin of AD patients showed a striking increase of eosinophil-associated CD48 expression while on peripheral blood leucocytes it was down-regulated. SA/exotoxins enhanced CD48 eosinophil expression, bound to CD48 and caused eosinophil activation and signal transduction. These effects were significantly decreased by blocking CD48 on human eosinophils or in BMEos from CD48-/- mice. We have also explored the role of CD48 in a SEB-induced peritonitis model in CD48-/- mice by evaluating inflammatory peritoneal cells, eosinophil numbers and activation.. These data demonstrate the important role of CD48 in SA/exotoxins-eosinophil activating interactions that can take place during allergic responses and indicate CD48 as a novel therapeutic target for allergy and especially of AD.

Topics: Animals; Antigens, CD; Bacterial Adhesion; Bone Marrow Cells; CD48 Antigen; Cell Degranulation; Dermatitis, Atopic; Enterotoxins; Eosinophils; Gene Expression; Humans; Interleukin-10; Interleukin-8; Leukocytes; Mice; Mice, Knockout; Peritonitis; Protein Binding; Signal Transduction; Skin; Staphylococcal Infections; Staphylococcus aureus

The article analyzes the results of surgical treatment of 78 patients with generalized peritonitis (GP) between the ages of 18 to 76 years. Severity was assessed by RP Mannheim peritoneal index (MPI). The study was conducted according to a comparative evaluation of the method of treatment in both groups. In the comparison group included 38 patients who received conventional therapy without immune complex. In 40 patients of the study group on the background of complex therapeutic measures in pre- and postoperative additionally used concomitant local and systemic ozone therapy (OT) with ozonatedperftoran (OP). All patients in the dynamics of blood prior to surgery, on the 3rd and 7th day after surgery was determined TNF-α, IL-4 and IL-8 by IFA. In general, patients in both groups at admission were identified imbalance between pro- and anti-inflammatory cytokines. Patients of the main group on the complex background of basic therapy combined local and systemic administration of OP positive effect in terms of acceleration available cytokine imbalance. The complex therapeutic interventions GP application of local and systemic OT with OP accelerates elimination of imbalances in the cytokine profile.

Topics: Administration, Intravenous; Adult; Aged; Blood Substitutes; Female; Fluorocarbons; Humans; Interleukin-4; Interleukin-8; Male; Middle Aged; Monitoring, Immunologic; Oxidants, Photochemical; Ozone; Perioperative Care; Peritoneal Lavage; Peritonitis; Severity of Illness Index; Treatment Outcome; Tumor Necrosis Factor-alpha

Meconium peritonitis is caused by an intestinal perforation that may occur in the fetus, followed by severe chemical peritonitis, resulting in high morbidity.. We have experienced six patients with meconium peritonitis. Cystic drainage was performed soon after birth for all patients. We investigated the concentrations of several cytokines and a chemokine (interleukin 8) in the ascites from the six patients with meconium peritonitis. In two patients we also measured the serum cytokines and chemokine level just after birth.. Interleukin 6 and interleukin 8 concentrations were very high in the cyst or ascites just after birth. In the serum taken from two patients, the levels of interleukin 6 and interleukin 8 were also high. In five patients who underwent drainage of cysts after birth, systemic inflammation could not be completely suppressed before curative surgery.. Interleukin 6 and interleukin 8 play important roles in the inflammatory response syndrome associated with meconium peritonitis, and drainage of cystic fluid did not completely suppress this inflammation. To lessen the high morbidity of meconium peritonitis, efforts should be made to suppress the inflammatory response using new treatment strategies, such as administration of steroids or anti-cytokine therapy to supplement cystic drainage.

Topics: Ascites; C-Reactive Protein; Chemokines; Cyst Fluid; Cytokines; Drainage; Fatal Outcome; Female; Fetal Diseases; Hernia, Diaphragmatic; Humans; Infant, Newborn; Interleukin-6; Interleukin-8; Intestinal Perforation; Male; Meconium; Peritonitis; Prognosis; Syndrome

To evaluate inflammatory/angiogenic cytokines-interleukin-1β (IL-1β), IL-6, IL-8, IL-12, interferon-γ (IFN-γ), tumor necrosis factor (TNF), and vascular endothelial growth factor A (VEGF-A)-in the peritoneal fluid of patients with endometriosis in relation to the occurrence and severity of pelvic adhesions and in control women without pelvic pathology.. Case-control study.. University research institution and hospital.. Sixty-five women with laparoscopically and histopathologically confirmed endometriosis, including 40 women with pelvic adhesions, and 37 control women without pelvic pathology.. Peritoneal fluid aspirated during routine diagnostic laparoscopic examination.. Cytokines evaluated in the peritoneal fluid via specific enzyme-linked immunosorbent assays.. Endometriosis and the revised American Fertility Society score of this disease were associated with statistically significantly increased levels of peritoneal IL-6 and IL-8 whereas the incidence and score of endometriosis-related pelvic adhesions were negatively associated with increased levels of VEGF-A. Notably, the concentration of VEGF-A predicted adhesion development and severity after adjustment for endometriosis severity. The adhesion score also correlated with increased levels of IL-6; however, after adjustment for endometriosis severity, the effect of this cytokine was no longer statistically significant.. Increased levels of VEGF-A may be associated with a decreased rate of pelvic adhesion formation in the course of endometriosis.

Topics: Ascitic Fluid; Case-Control Studies; Cytokines; Endometriosis; Female; Humans; Interferon-gamma; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Peritonitis; Severity of Illness Index; Tissue Adhesions; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

The aim of this study was to investigate the effects of carbon dioxide (CO(2)) pneumoperitoneum on the intra-abdominal spread of bacteria, the local and systemic cytokine expression, and oxidant/antioxidant status in young rats with bacterial peritonitis.. Young Sprague-Dawley rats, aging 20-27 days and weighing around 50 g, were allocated to six groups of six to nine animals in each. Intra-abdominal infection model was developed by intraperitoneal injection with 1 cc of Escherichia coli (E. coli) (10(8) CFU/mL, ATCC25922 strain) via right lower abdominal wall. Carbon dioxide (CO(2)) pneumoperitoneum was applied to the rats via umbilical pit insufflation with 20 cc CO(2) for 30 min. All survived rats underwent laparotomy and were killed 24 h or 3 days later. Serum levels of CO(2) and CRP were measured. Left lower abdomen peritoneum, peritoneal fluid, mesenteric lymph node of terminal ileum, and liver were taken for bacterial culture. Liver and plasma levels of TNF-α, IL-1β, and IL-6 were examined for the level of local and systemic immunologic response. Oxidant/antioxidant status in liver and plasma were assessed by measuring malondialdehyde (MDA), and reduced to oxidized glutathione ratio (GSH/GSSG).. Carbon dioxide (CO(2)) pneumoperitoneum does not facilitate E. coli dissemination to other intra-abdominal organs in rats with localized E. coli peritonitis. Peritonitis rats that underwent abdominal CO(2) insufflation have insignificantly higher CRP or lower CO(2) levels. Plasma and liver TNF-α, IL-1β concentrations were not significantly different among the four groups, but plasma IL-6 was significantly increased in rats with E. coli peritonitis and CO(2) pneumoperitoneum that were killed 3 days later as compared with that of rats that were killed 24 h later. In rats with E. coli peritonitis, CO(2) pneumoperitoneum was significantly associated with decreased hepatic GSH/GSSG ratio. However, plasma and liver MDA levels were not altered after CO(2) pneumoperitoneum.. Carbon dioxide (CO(2)) pneumoperitoneum is not associated with E. coli dissemination in the presence of local intra-abdominal infection. CO(2) pneumoperitoneum elicited systemic anti-inflammatory response at a specific time period and decreased hepatic antioxidant status in young rats with E. coli peritonitis.

Topics: Analysis of Variance; Animals; C-Reactive Protein; Carbon Dioxide; Escherichia coli Infections; Glutathione; Insufflation; Interleukin-6; Interleukin-8; Liver; Malondialdehyde; Oxidative Stress; Peritonitis; Pneumoperitoneum; Rats; Rats, Sprague-Dawley; Statistics, Nonparametric; Tumor Necrosis Factor-alpha

5-Lipoxygenase (5-LOX) plays key roles in infection and allergic responses. Herein, four 5-LOX-derived lipids comprising 5-hydroxyeicosatetraenoic acid (HETE) attached to phospholipids (PLs), either phosphatidylethanolamine (PE) or phosphatidylcholine (18:0p/5-HETE-PE, 18:1p/5-HETE-PE, 16:0p/5-HETE-PE, and 16:0a/5-HETE-PC), were identified in primary human neutrophils. They formed within 2 minutes in response to serum-opsonized Staphylococcus epidermidis or f-methionine-leucine-phenylalanine, with priming by lipopolysaccharide, granulocyte macrophage colony-stimulating factor, or cytochalasin D. Levels generated were similar to free 5-HETE (0.37 ± 0.14 ng vs 0.55 ± 0.18 ng/10(6) cells, esterified vs free 5-HETE, respectively). They remained cell associated, localizing to nuclear and extranuclear membrane, and were formed by fast esterification of newly synthesized free 5-HETE. Generation also required Ca(2+), phospholipase C, cytosolic and secretory phospholipase A(2), 5-LOX activating protein, and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1. 5-HETE-PLs were detected in murine S epidermidis peritonitis, paralleling neutrophil influx, and in effluent from Gram-positive human bacterial peritonitis. Formation of neutrophil extracellular traps was significantly enhanced by 5-LOX inhibition but attenuated by HETE-PE, whereas 5-HETE-PE enhanced superoxide and interleukin-8 generation. Thus, new molecular species of oxidized PL formed by human neutrophils during bacterial infection are identified and characterized.

Topics: Aged; Aged, 80 and over; Animals; Arachidonate 5-Lipoxygenase; Bacterial Infections; Eicosanoids; Female; Gram-Positive Bacterial Infections; Humans; Hydroxyeicosatetraenoic Acids; In Vitro Techniques; Interleukin-8; Male; Mice; Mice, Inbred C57BL; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Peritonitis; Phospholipids; Plasmalogens; Signal Transduction; Staphylococcal Infections; Staphylococcus epidermidis; Superoxides; Tandem Mass Spectrometry; Tetradecanoylphorbol Acetate

Human blood Vγ9/Vδ2 T cells, monocytes and neutrophils share a responsiveness toward inflammatory chemokines and are rapidly recruited to sites of infection. Studying their interaction in vitro and relating these findings to in vivo observations in patients may therefore provide crucial insight into inflammatory events. Our present data demonstrate that Vγ9/Vδ2 T cells provide potent survival signals resulting in neutrophil activation and the release of the neutrophil chemoattractant CXCL8 (IL-8). In turn, Vγ9/Vδ2 T cells readily respond to neutrophils harboring phagocytosed bacteria, as evidenced by expression of CD69, interferon (IFN)-γ and tumor necrosis factor (TNF)-α. This response is dependent on the ability of these bacteria to produce the microbial metabolite (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP), requires cell-cell contact of Vγ9/Vδ2 T cells with accessory monocytes through lymphocyte function-associated antigen-1 (LFA-1), and results in a TNF-α dependent proliferation of Vγ9/Vδ2 T cells. The antibiotic fosmidomycin, which targets the HMB-PP biosynthesis pathway, not only has a direct antibacterial effect on most HMB-PP producing bacteria but also possesses rapid anti-inflammatory properties by inhibiting γδ T cell responses in vitro. Patients with acute peritoneal-dialysis (PD)-associated bacterial peritonitis--characterized by an excessive influx of neutrophils and monocytes into the peritoneal cavity--show a selective activation of local Vγ9/Vδ2 T cells by HMB-PP producing but not by HMB-PP deficient bacterial pathogens. The γδ T cell-driven perpetuation of inflammatory responses during acute peritonitis is associated with elevated peritoneal levels of γδ T cells and TNF-α and detrimental clinical outcomes in infections caused by HMB-PP positive microorganisms. Taken together, our findings indicate a direct link between invading pathogens, neutrophils, monocytes and microbe-responsive γδ T cells in early infection and suggest novel diagnostic and therapeutic approaches.

Topics: Adult; Antigen-Presenting Cells; Bacteria; Bacterial Infections; Cell Communication; Cell Differentiation; Cell Survival; Cells, Cultured; Diphosphates; Humans; Interferon-gamma; Interleukin-8; Lymphocyte Activation; Monocytes; Neutrophil Activation; Neutrophils; Peritonitis; Phagocytosis; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocyte Subsets; Tumor Necrosis Factor-alpha

PMN leukocytes are the most abundant leukocytes in the circulation and play an important role in host defense. PMN leukocyte recruitment and inflammatory responses at sites of infection are critical components in innate immunity. Although inflammation and coagulation are known to have bidirectional relationships, little is known about the interaction between PMN leukocytes and coagulation factors. Coagulation FXI participates in the intrinsic coagulation pathway upon its activation, contributing to hemostasis and thrombosis. We have shown previously that FXI-deficient mice have an increased survival and less leukocyte accumulation into the peritoneum in severe polymicrobial peritonitis. This result suggests a role for FXI in leukocyte trafficking and/or function. In this study, we characterized the functional consequences of FXIa binding to PMN leukocytes. FXIa reduced PMN leukocyte chemotaxis triggered by the chemokine, IL-8, or the bacterial-derived peptide, fMLP, perhaps as a result of the loss of directed migration. In summary, our data suggest that FXIa modulates the inflammatory response of PMN leukocytes by altering migration. These studies highlight the interplay between inflammation and coagulation and suggest that FXIa may play a role in innate immunity.

Topics: Blood Coagulation; Cell Movement; Cells, Cultured; Chemotaxis, Leukocyte; Factor XIa; Humans; Inflammation; Interleukin-8; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Peritonitis; Protein Binding

Migration and activation of polymorphonuclear neutrophils (PMN) and apoptosis are central to inflammatory tissue damage. This study examines the relation of these processes, and their expression in the abdominal, systemic, and bronchoalveolar compartments in patients with severe peritonitis.. Thirty-one consecutive patients undergoing laparotomy for severe secondary peritonitis. Eight operated patients without peritonitis and 10 long-term mechanically ventilated noninfected patients served as controls. Peritoneal fluid, blood, and bronchoalveolar lavage fluid (BALF) was obtained on d 0 (day of initial laparotomy), 2, and 3. Levels of chemokines (interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1), PMN-counts, PMN activation [myeloperoxidase (MPO), elastase] and apoptosis (nucleosomes) were determined.. In peritonitis patients, levels of chemokines and markers of PMN sequestration were increased in all compartments. IL-8 levels were higher in BALF than in plasma, and did not originate from the circulation or from lysis of alveolar cells. Pulmonary nucleosome levels were higher in patients who died (P=0.020), and corresponded with PMN-count in BALF (P<0.001), levels of chemokines (IL-8, P=0.003; MCP-1, P=0.001), and PMN-activation (MPO, P<0.001; elastase P=0.007).. Severe peritonitis produces an early pulmonary expression of chemoattractants creating a gradient for PMN sequestration and activation into the lung. These parameters are associated with expression of apoptosis in the lung, which is increased in nonsurviving peritonitis patients.

Topics: Abdomen; Aged; APACHE; Apoptosis; Bronchoalveolar Lavage Fluid; Chemokine CCL2; Chemotaxis; Female; Humans; Interleukin-8; Length of Stay; Leukocyte Count; Male; Middle Aged; Neutrophils; Nucleosomes; Peritonitis; Peroxidase; Positive-Pressure Respiration

Sepsis and septic shock are frequently encountered in the intensive care unit. Despite the evolution of intensive care medicine during the last decades, septic shock is still associated with high mortality and complications of sepsis such as cholestasis, liver dysfunction, and massive intravascular volume deficit. Little is known about the whole pattern of changes at the transcriptional level during the development of acute sepsis. Here we present a detailed molecular biological analysis of the events in the liver during the first day of acute bacterial infection in a clinically relevant model of porcine peritoneal sepsis. Before and 21 h after induction of sepsis by autologous fecal inoculum, liver samples were taken for microarray analysis. There were two groups of animals (7 control and 8 sepsis), two of each group where used in microarray, the remaining were used for confirmation of selected genes by real-time polymerase chain reaction. Pathway analysis revealed that in acute sepsis, gene expression was significantly changed in processes related to apoptosis, inflammation, and oxidant/redox balance. Although after 21 h these animals are expected to die within the next 3 to 4 h from massive complications, functional induction of apoptosis could not be confirmed. Computer analysis identified three key regulator genes (IL8, CCL2, and CXCL2) among the first genes to be upregulated specifically in the sepsis group, and these can directly or indirectly control the bulk of the sepsis response. Induction of inflammatory mediators by sepsis was supported by the detection of corresponding cytokines (interleukin 6 and interleukin 8) in the blood.

Topics: Animals; Chemokine CCL2; Chemokine CXCL2; Fluid Therapy; Interleukin-8; Liver; Oligonucleotide Array Sequence Analysis; Peritonitis; Resuscitation; Sepsis; Swine

Polymorphonuclear leukocyte (PMN) infiltration is a cardinal feature of peritonitis. CXC chemokine ligands 1 and 8 (CXCL1 and CXCL8), and the cytokine granulocyte colony-stimulating factor (G-CSF) are the key mediators of PMN accumulation. Increasing evidence points to an important role of human peritoneal fibroblasts (HPFB) in the response of the peritoneum to infection. We have examined the synthesis of PMN-targeting cytokines by HPFB exposed to intraperitoneal milieu as represented by peritoneal dialysate effluent (PDE) from patients undergoing peritoneal dialysis. PDE obtained during peritonitis, but not during infection-free periods, significantly increased production of CXCL1, CXCL8, and G-CSF by HPFB. The effect was largely blocked by antibodies to interleukin-1beta (IL-1beta), whereas neutralization of tumor necrosis factor-alpha (TNFalpha) had no major effect. Similar pattern of inhibition was observed when HPFB were exposed to conditioned media from endotoxin-stimulated peritoneal macrophages. Significance of IL-1beta stimulation was further shown in experiments with recombinant cytokines. Compared with TNFalpha, exposure of HPFB to recombinant IL-1beta resulted in a much higher release of PMN-targeting cytokines. The assessment of mRNA degradation revealed that the IL-1beta-induced transcripts of CXCL1, CXCL8, and G-CSF were more stable compared with those induced by TNFalpha. These data indicate that HPFB can be a significant source of PMN-targeting cytokines when stimulated with IL-1beta in the inflamed peritoneum.

Topics: Adult; Aged; Cells, Cultured; Chemokine CXCL1; Female; Fibroblasts; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-1beta; Interleukin-8; Macrophages, Peritoneal; Male; Middle Aged; Peritoneal Cavity; Peritoneal Dialysis; Peritonitis; Recombinant Proteins; Tumor Necrosis Factor-alpha; Young Adult

To explore the characteristics of immune imbalance in patients with multiple organ dysfunction syndrome (MODS) induced by severe intra-abdominal infection and its relationship with changing of TCM sthenia-asthenia syndrome.. Forty-six patients with MODS induced by severe intra-abdominal infection and treated with etiological and syndrome differentiation of integrative medicine were observed in succession. Patients' peripheral blood levels of interleukin-6/interleukin-10 ratio (IL-6/IL-10), human leukocyte antigen DR site (HLA-DR), helper T lymphocyte1/2 ratio (Th1/Th2), and the regulatory T lymphocyte (Treg) were measured on the 1st, 3rd and 7th day of the research respectively. And the distribution laws of TCM syndrome types, sthenia (S), asthenia (A), and mingled sthenia/asthenia (M), in patients were observed as well.. IL-6/IL-10 ratio at all the testing time points showed insignificant difference in patients of types S and M, while in those of type A, it was more lowered on the 7th day than that on the 1st day. HLA-DR lowered to <30% on the 7th day in all patients of type A and showed significant difference to that on the 1st day (P <0.05), while HLA-DR <30% was not found in all patients of types S and M. Th1/Th2 ratio in patients of types S and A was insignificant different at the foremost 3 days, but lowered significantly on the 7th day, while in patients of type M, it was unchanged in all the 7 days of observation. Treg level was unchanged in the foremost 3 days in patients of types S and M, while in those of type A, it raised on the 3rd day, but no raising was found in the subsequent 4 days. Comparisons of various indexes detected at corresponding time points respectively among patients with various syndrome types showed that, for levels of IL-6/IL-8, HLA-DR, and Th1/Th2, the sequence was S>M>A; and for Treg, it was A>M>S.. In the pathological process of MODS induced by severe intra-abdominal infection, the index IL-6/IL-10, reflecting the balance of the pro-/anti-inflammatory cytokines and the indexes HLA-DR, Th1/Th2 and Treg reflecting the immune function, all can exactly reflect the TCM asthenia-sthenia syndrome types. The sequence in patients of various syndrome types for levels of IL-6/IL-10, HLA-DR and Th1/Th2, is S> M>A, but for Treg it is the inverse, as A>M>S.

Topics: Adolescent; Adult; Aged; Diagnosis, Differential; HLA-DR Antigens; Humans; Interleukin-6; Interleukin-8; Medicine, Chinese Traditional; Middle Aged; Multiple Organ Failure; Peritonitis; Sepsis; T-Lymphocytes, Regulatory; Th1 Cells; Th2 Cells; Yang Deficiency; Young Adult

A series of 6-amino-4-oxo-1,3-diphenyl-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-carbonyl derivatives was synthesized. The compounds demonstrated to be novel, potent and selective inhibitors of Interleukin-8-induced human neutrophil chemotaxis. A SAR study was performed by varying the carbonyl function at position 5 and the chain linked to the amino group at position 6 of the scaffold. All the compounds of the series displayed inhibitory activity at nano- or picomolar concentrations against Interleukin-8-driven migration and no activity against fMLP- and C5a-induced chemotaxis. The binding tests of selected compounds on CXCR1 and CXCR2 receptors were negative. The most potent derivative showed in vivo efficacy in a mouse model of Zymosan-induced peritonitis.

Topics: Animals; Carboxylic Acids; Chemotaxis, Leukocyte; Disease Models, Animal; Interleukin-8; Mice; Neutrophils; Peritonitis; Pyrimidines; Structure-Activity Relationship

Interleukin (IL)-22 production triggered by innate immune mechanisms has been identified as key to efficient intestinal anti-bacterial host defence and preservation of homeostasis. We hypothesized that glucocorticoid therapy may impair IL-22 expression, which should promote intestinal epithelial damage with the potential of subsequent bacterial translocation. High-dose corticosteroid therapy in Crohn's disease has been associated with an increased rate of abscess formation and ultimately with a higher risk of developing postoperative infectious complications, including abdominal sepsis. Thus, we sought to investigate effects of the prototypic glucocorticoid dexamethasone on IL-22 production in the context of bacterial infection. Enhanced IL-22 plasma levels were detectable in rat sepsis. Moreover, heat-inactivated Staphylococcus epidermidis, used as a prototypic activator of innate immunity, induced robust production of IL-22 by human peripheral blood mononuclear cells (PBMC). Here, we report for the first time that dexamethasone mediates remarkable suppression of IL-22 as detected in S. epidermidis-activated PBMC and rat sepsis, respectively. The data presented herein suggest that insufficient IL-22 function may contribute to impaired intestinal host defence in the context of corticosteroid therapy.

Topics: Animals; Case-Control Studies; Cells, Cultured; Depression, Chemical; Dexamethasone; Gene Expression; Glucocorticoids; Humans; Interleukin-10; Interleukin-22; Interleukin-8; Interleukins; Male; Models, Animal; Peritonitis; Random Allocation; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Staphylococcal Infections; Staphylococcus epidermidis

In inflammatory diseases, circulating neutrophils are recruited to sites of injury. Attractant signals are provided by many different chemotactic molecules, such that blockade of one may not prevent neutrophil recruitment effectively. The Slit family of secreted proteins and their transmembrane receptor, Robo, repel axonal migration during CNS development. Emerging evidence shows that by inhibiting the activation of Rho-family GTPases, Slit2/Robo also inhibit migration of other cell types toward a variety of chemotactic factors in vitro and in vivo. The role of Slit2 in inflammation, however, has been largely unexplored. We isolated primary neutrophils from human peripheral blood and mouse bone marrow and detected Robo-1 expression. Using video-microscopic live cell tracking, we found that Slit2 selectively impaired directional migration but not random movement of neutrophils toward fMLP. Slit2 also inhibited neutrophil migration toward other chemoattractants, namely C5a and IL-8. Slit2 inhibited neutrophil chemotaxis by preventing chemoattractant-induced actin barbed end formation and cell polarization. Slit2 mediated these effects by suppressing inducible activation of Cdc42 and Rac2 but did not impair activation of other major kinase pathways involved in neutrophil migration. We further tested the effects of Slit2 in vivo using mouse models of peritoneal inflammation induced by sodium periodate, C5a, and MIP-2. In all instances, Slit2 reduced neutrophil recruitment effectively (P<0.01). Collectively, these data demonstrate that Slit2 potently inhibits chemotaxis but not random motion of circulating neutrophils and point to Slit2 as a potential new therapeutic for preventing localized inflammation.

Topics: Animals; cdc42 GTP-Binding Protein; Cell Polarity; Chemokine CXCL2; Chemotaxis; Complement C5a; Disease Models, Animal; Enzyme Activation; Gene Expression Regulation; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Mice; N-Formylmethionine Leucyl-Phenylalanine; Nerve Tissue Proteins; Neutrophils; Peritonitis; rac GTP-Binding Proteins; RAC2 GTP-Binding Protein; Receptors, Immunologic; Roundabout Proteins

Neutrophils chemotaxis is a complex multistep process that, if upregulated, causes acute inflammation and a number of autoimmune diseases. We report here the synthesis of a new N-(4-substituted)pyrazolyl-N'-alkyl/benzyl/phenylureas that are potent inhibitors of interleukin-8 (IL8)-induced neutrophil chemotaxis. The first series of compounds, obtained by functionalization with a urea moiety of the 5-amino-1-(2-hydroxy-2-phenylethyl)-1H-pyrazole-4-carboxylic acid ethyl ester 3, blocked the IL8-induced neutrophil chemotaxis, while they did not block N-formylmethionylleucylphenylalanine-mediated chemotaxis. The most active compounds, 3-benzyl- (4d), 3-(4-benzylpiperazinyl)- (4i), 3-phenyl- (4k) and 3-isopropylureido (4a) derivatives, showed an IC50 of 10, 14, 45, and 55 nM, respectively. Several different molecules were then synthesized to obtain more information for SAR study. Compounds 4a, 4d, and 4k were inactive in the binding assays on CXCR1 and CXCR2 (IL8 receptors), whereas they inhibited the phosphorylation of PTKs (protein tyrosine kinases) in the 50-70 kDa region. Moreover, in the presence of the same derivatives, we observed a complete block of F-actin rise and pseudopod formation.

Topics: Actins; Adult; Animals; Anti-Inflammatory Agents; Chemotaxis, Leukocyte; Humans; Interleukin-8; Male; Mice; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Peritoneal Cavity; Peritonitis; Phenylurea Compounds; Phosphorylation; Proto-Oncogene Proteins c-akt; Pseudopodia; Pyrazoles; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Structure-Activity Relationship

Human chymase induced release of interleukin-8 (IL-8) in human EoL-1 cells that had been differentiated into eosinophil-like cells with butyric acid. The chymase-induced IL-8 production was specific in that other cytokines/chemokines examined were not induced. Human chymase also increased mRNA for IL-8 in the differentiated EoL-1 cells, showing involvement of mRNA synthesis. The chymase-induced IL-8 release was inhibited by pertussis toxin as well as U0126 (an inhibitor for extracellular signal-regulated kinase pathway) and SB203580 (p38 inhibitor), suggesting that the chymase-induced IL-8 production is mediated by G protein-coupled receptor and mitogen-activated protein kinases. Mouse mast cell protease-4 (mMCP-4), a mouse chymase, induced macrophage-inflammatory protein-2 (MIP-2), a mouse homologue for IL-8, in mouse eosinophils in vitro. Intradermal injection of mMCP-4 not only induced skin edema but increased MIP-2 content and neutrophil number at the injection site. Taken together, our findings demonstrate that mast cell chymase may contribute to the interaction between eosinophils and neutrophils by inducing IL-8/MIP-2 in eosinophils at allergic inflamed sites.

Topics: Animals; Butadienes; Cell Line, Tumor; Chemokine CXCL2; Chemokines; Chymases; Cytokines; Dose-Response Relationship, Drug; Enzyme Inhibitors; Eosinophils; Gene Expression; Humans; Imidazoles; Interleukin-8; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Neutrophils; Nitriles; Peritonitis; Pertussis Toxin; Pyridines; Quinazolines; RNA, Messenger; Serine Endopeptidases

The predominant problems associated with peritoneal dialysis (PD) are ultrafiltration failure and peritonitis. PD maintains a state of intraperitoneal inflammation that affects the structure and function of the peritoneal membrane, potentially impairing ultrafiltration efficiency. Paradoxically, some PD fluids also have anti-inflammatory properties that may compromise the immune defense against peritonitis. This anti-inflammatory feature is mostly due to the glucose degradation products (GDPs), formed during heat-sterilization and storage of PD fluids. The main purpose of the present thesis was to study regulatory mechanisms behind the acute intraperitoneal inflammatory response in PD in the presence and absence of experimental peritonitis. Rats were exposed to a single dose of heat- or filter sterilized PD fluids either as an i.p. injection or as an infusion through an indwelling catheter, with or without supplementations, or pretreatment of the animals. The dwell fluid was analyzed zero, two and four hours later concerning activation of the complement and coagulation cascades, neutrophil recruitment and respiratory burst, ultrafiltration volumes, cytokine-induced neutrophil chemoattractant (CINC-1), rat mast cell protease 2 (RMCP-2), glucose, urea and histamine concentrations and ex vivo/in vitro intraperitoneal chemotactic activity. Exposure to filter sterilized PD fluid alone induced intraperitoneal complement activation and coagulation, neutrophil recruitment and increased the levels of CINC-1 during the dwell. Intraperitoneal concentrations of the mast cell markers histamine and RMCP-2 changed little during the dwells and did not indicate mast cell activation. Low molecular weight heparin (LMWH) and C5 blockade improved ultrafiltration. Pretreatment with cobra venom factor, known decomplementing agent, blocked the CINC-1 release and the neutrophil recruitment and improved ultrafiltration. In combination with experimental peritonitis, heat sterilized PD fluid compared to filter sterilized, inhibited the CINC-1 release and the recruitment of neutrophils to the peritoneal cavity without affecting the intraperitoneal complement activation. The results of the present thesis indicate that addition of LMWH to the PD fluid improves ultrafiltration, probably by blocking C5a activity. C5 blockade seems to improve ultrafiltration by a mechanism that involves a reduction in glucose transport, possibly by reducing C5 induced vasodilatation. Complement activation

Topics: Acute Disease; Animals; Anti-Inflammatory Agents; Anticoagulants; Blood Coagulation; Chemokine CXCL1; Chemokines, CXC; Complement Activation; Complement C5a; Dialysis Solutions; Heparin, Low-Molecular-Weight; Intercellular Signaling Peptides and Proteins; Interleukin-8; Male; Mast Cells; Neutrophil Infiltration; Peritoneal Dialysis; Peritonitis; Rats; Rats, Sprague-Dawley; Respiratory Burst

The aim of the study was to investigate interleukin 8 (IL-8) mRNA gene expression in circulating and emigrated intra-abdominal neutrophils during human secondary peritonitis intra- and postoperatively (until 96 h). Patients with secondary peritonitis were allocated to two groups, e.g., patients with no complications (n = 10) and patients with complications (organ failure, septic shock, etc., n = 9). Patients with elective abdominal surgery (n = 11) and a group with healthy volunteers (n = 7) were studied as controls. Neutrophil RNA was isolated and semiquantitative reverse transcription-PCR was performed. The PCR products were compared with corresponding GAPDH bands (=100%). The highest amount of IL-8 mRNA could be assessed in blood neutrophils of healthy volunteers (87.4% +/- 7.4%). Complicated peritonitis was associated with the lowest concentration of IL-8 mRNA in blood neutrophils intraoperatively (24% +/- 7%, P < 0.05), which showed no recovery throughout the observation period (34% +/- 8%, 96 h postoperatively). IL-8 mRNA concentration in blood neutrophils of patients with uncomplicated peritonitis and patients with elective abdominal surgery was higher intraoperatively (55.2% +/- 9% (uncomplicated peritonitis); 68% +/- 15% (elective abdominal surgery, P < 0.05 versus complicated peritonitis). Thereafter, IL-8 mRNA decreased slightly in both groups, but was distinctly higher than in patients with complicated peritonitis. Emigration to the abdominal cavity resulted in an approximately 2-fold, in some cases 3-fold, increase in the concentration of IL-8 mRNA in emigrated intra-abdominal neutrophils when compared with circulating cells. This increase could be observed in all groups. The long-lasting down-regulation of constitutive gene expression of IL-8 mRNA in blood neutrophils during complicated peritonitis is worrying because IL-8 is an important activator and chemoattractant for neutrophils themselves. It is encouraging that migration to another compartment, e.g., infected abdominal cavity, resulted in an increase in neutrophil IL-8 mRNA during complicated and uncomplicated peritonitis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged; Neutrophils; Peritoneum; Peritonitis; Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Time Factors

Chronic alcoholic patients have a threefold to fourfold increased risk for developing a severe infection or septic shock after surgery, which might be due to altered immune response. The aim of this outcome matched study was to investigate proinflammatory and anti-inflammatory immune parameters during the course of infection and subsequent septic shock in chronic alcoholic patients, and to compare these parameters with those in nonalcoholic patients.. Twenty-eight patients from a cohort of fifty-six with either pneumonia or peritonitis and subsequent septic shock were selected. Fourteen patients were chronic alcoholics whereas fourteen were nonalcoholic patients. Chronic alcoholic patients met criteria (Diagnostic and Statistical Manual of Mental Disorders IV, of the American Psychiatric Association) for alcohol abuse or dependence. Measurements were performed during the onset of infection (within 24 hours after the onset of infection), in early septic shock (within 12 hours after onset of septic shock) and in late septic shock (72 hours after the onset). Blood measurements included proinflammatory and anti-inflammatory cytokines.. Chronic alcoholic patients exhibited significantly lower plasma levels of IL-8 (P < 0.010) during the onset of infection than did matched nonalcoholic patients. In early septic shock, chronic alcoholic patients had significantly decreased levels of IL-1beta (P < 0.015), IL-6 (P < 0.016) and IL-8 (P < 0.010). The anti-inflammatory parameters IL-10 and tumour necrosis factor receptors I and II did not differ between alcoholic and nonalcoholic patients.. At the onset of infection and during early septic shock, chronic alcoholic patients had lower levels of proinflammatory immune parameters than did nonalcoholic patients. Therefore, immunomodulatory therapy administered early may be considered in chronic alcoholic patients at the onset of an infection because of their altered proinflammatory immune response.

Topics: Adult; Aged; Alcoholism; Case-Control Studies; Chronic Disease; Humans; Interleukin-1; Interleukin-10; Interleukin-8; Middle Aged; Peritonitis; Pneumonia; Predictive Value of Tests; Receptors, Tumor Necrosis Factor; Risk Factors; Shock, Septic; Tumor Necrosis Factor-alpha

Aim of the study was to characterize different functions of circulating and emigrated, intra-abdominal polymorphonuclear leukocytes (cPMNs, ePMNs) during human secondary peritonitis.. In patients (n=25) with diffuse secondary peritonitis circulating and emigrated PMNs were characterized intra- and until 96 h postoperatively. Patients were allocated to two different groups, e. g. patients with septic complications (shock, organ failure, n=11) and patients without complications (n=14) during peritonitis. In addition a control group of patients (n=10) with abdominal surgery but without peritonitis was investigated. The lucigenin- and luminol-enhanced chemiluminescence was used to determine extra- and intracellular oxygen radical generation of PMNs. Besides spontaneous oxygen radical generation of PMNs, stimulated radical production was investigated after the addition of ionophores A23 187 and C3-coated zymosan. Phagocytosis by PMNs was characterized with opsonized E. coli bacteria and fluorescence-activated cell analysis.. Especially patients with complicated peritonitis had strong and long-lasting changes of PMNs functions. The toxic and tissue-destroying production of extracellular oxygen radicals by circulating PMNs was enhanced (e. g., A23 187 - stimulated oxygen radical generation 433 +/- 89 cpm/cPMNs (peritonitis with complications) versus 90 +/- 30 cpm/cPMNs (peritonitis without complications) versus 110 +/- 44 cpm/cPMNs (controls), p < 0.05). Phagocytosis (58 +/- 9 % (ePMNs, peritonitis with complications) versus 81 +/- 6 % (ePMNs, peritonitis without complications) versus 82.2 +/- 1.6 % (ePMNs, controls), p < 0.05) and phagocytosis-associated intracellular oxygen radical generation (8.23 +/- 1.6 x 10(3) cpm/ePMNs (peritonitis with complications) versus 25.2 +/- 5.2 x 10(3) cpm/ePMNs (peritonitis without complications) versus 11.7 +/- 2.8 cpm x 10(3) cpm/ePMNs (controls) p < 0.05) were suppressed.. Not for all patients with peritonitis does it seem favourable to modulate PMNs-functions. If immunomodulation would be able to down-regulate exaggerated functions of circulating PMNs and to up-regulate the suppressed functions of emigrated PMNs patients with complicated peritonitis might benefit from this therapy.

Topics: Colectomy; Free Radicals; Gastrectomy; Humans; Interleukin-8; Luminescent Measurements; Multiple Organ Failure; Neutrophils; Peritoneum; Peritonitis; Postoperative Complications; Prognosis; Reactive Oxygen Species; Shock, Septic; Tumor Necrosis Factor-alpha

Galectin-1 (Gal-1), the prototype of a family of beta-galactoside-binding proteins, has been shown to attenuate experimental acute and chronic inflammation. In view of the fact that endothelial cells (ECs), but not human polymorphonuclear leukocytes (PMNs), expressed Gal-1 we tested here the hypothesis that the protein could modulate leukocyte-EC interaction in inflammatory settings. In vitro, human recombinant (hr) Gal-1 inhibited PMN chemotaxis and trans-endothelial migration. These actions were specific as they were absent if Gal-1 was boiled or blocked by neutralizing antiserum. In vivo, hrGal-1 (optimum effect at 0.3 micro g equivalent to 20 pmol) inhibited interleukin-1beta-induced PMN recruitment into the mouse peritoneal cavity. Intravital microscopy analysis showed that leukocyte flux, but not their rolling velocity, was decreased by an anti-inflammatory dose of hrGal-1. Binding of biotinylated Gal-1 to resting and postadherent human PMNs occurred at concentrations inhibitory in the chemotaxis and transmigration assays. In addition, the pattern of Gal-1 binding was differentially modulated by PMN or EC activation. In conclusion, these data suggest the existence of a previously unrecognized function of Gal-1, that is inhibition of leukocyte rolling and extravasation in experimental inflammation. It is possible that endogenous Gal-1 may be part of a novel anti-inflammatory loop in which the endothelium is the source of the protein and the migrating PMNs the target for its anti-inflammatory action.

Topics: Animals; Binding Sites; Cell Communication; Cell Movement; Chemotaxis, Leukocyte; Dose-Response Relationship, Drug; Endothelium, Vascular; Flow Cytometry; Galectin 1; Humans; In Vitro Techniques; Injections; Interleukin-1; Interleukin-8; Leukocyte Rolling; Male; Mice; Neutrophils; Peritonitis; Recombinant Proteins

The aim of this study was to evaluate the influence of taurolidine (TAU) and polyhexanid (POLY) on basic inflammatory reactions during peritonitis by using an in vitro model of human peritoneum.. Human umbilical vein endothelial cells (HUVEC) and human peritoneal mesothelial cells (HPMC; concentration: 2x10(5)/cm2) were brought on a collagen-coated filter insert with 3-microm pore size (HUVEC on the bottom, HPMC on the top), thus resulting in a two-chamber peritoneal model. After 5 days, confluence of the cells was reached, and HPMC were stimulated with 0.5 mL of TNF-alpha (10 microg/mL) for 4 h. Afterwards, 0.5 mL of TAU (1% and 2%) or 0.5 mL of POLY (0.1% and 0.2%) solution were added to the upper (HPMC) compartment. Polymorphonuclear neutrophils (PMN, 10(6)/mL) were placed in the lower compartment 1 h later. After 2 and 6 h, aliquots were taken from the upper compartment and transmigrated PMN were counted. Interleukin-8 (IL-8) concentrations were measured in both compartments by chemiluminescent enzyme immunometric assay. Expression of the adhesion molecules P-selectin and intercellular adhesion molecule-1 (ICAM-1) was assessed by immunohistochemistry. Controls were either TNF-alpha-stimulated HPMC without any antiseptic agents, or stimulated HPMC where TNF-alpha had been substituted by culture medium. Each experiment was performed in triplicate.. Stimulation with TNF-alpha led to a time-dependent increase of IL-8 secretion to the apical compartment resulting in a gradient between both chambers, as well as to a time-dependent increase of PMN transmigration and expression of adhesion molecules. IL-8 gradients and PMN migration were significantly higher as compared to the other groups (p<0.05). After substitution of the stimulus by culture medium, significantly less IL-8 was measured in both compartments. PMN transmigration was almost absent (p<0.05). Addition of POLY and TAU led to comparable low IL-8 gradients with concomitant low PMN transmigration. The initially detected expression of adhesion molecules significantly decreased during the observation time. The IL-8 gradient in all groups correlated significantly with PMN transmigration (r=0.74226; p<0.0001).. The diminished IL-8 response together with low PMN transmigration rates after addition of TAU and POLY may reflect either antiinflammatory effects or cellular damage.

Topics: Anti-Infective Agents, Local; Anti-Inflammatory Agents; Biguanides; Cell Movement; Dose-Response Relationship, Drug; Endothelial Cells; Endothelium, Vascular; Humans; In Vitro Techniques; Intercellular Adhesion Molecule-1; Interleukin-8; Neutrophils; P-Selectin; Peritoneal Cavity; Peritonitis; Taurine; Thiadiazines; Time Factors; Tumor Necrosis Factor-alpha

To study the changes in solute transperitoneal transport and the IL-8 level at different time phases in dialysis-related peritonitis so as to understand the mechanism of solute transperitoneal transport.. Forty-two New Zealand white rabbits were randomly divided into three experimental groups (5 min, 24 h, and 48 h groups) and a control group. The rabbit model of peritoneal dialysis-related peritonitis was created. The plasma and effluent concentration of creatinine, glucose, total proteins, and albumin were determined respectively; the D/P values of creatinine, total proteins, and albumin, and the Dn/D0 values of glucose were calculated respectively.. There was a significant difference in the D/P values of albumin at 120, 240, and 360 min between the 48 h group and the control group (P < 0.05); the D/P values of total proteins significantly increased at different time points in the 48 h group compared with the control group (P < 0.05); there was a significant difference in the creatinine D/P values and glucose Dn/D0 values between the 48 h group and the control group (P < 0.05). Compared with the control group, IL-8 and WBC obviously increased in the 48 h group (P < 0.05); the effluent IL-8 levels at different time points were positively correlated to the D/P values of the total proteins (P < 0.05). The effluent IL-8 levels at different time points after 60 min were positively correlated to the D/P values of albumin (P < 0.05).. The mechanism of transperitoneal transport of the different molecule solutes varies. Both IL-8 and WBC can influence solute transperitoneal transport in peritoneal dialysis-related peritonitis.

Topics: Animals; Interleukin-8; Peritoneal Dialysis; Peritoneum; Peritonitis; Rabbits; Random Allocation

Although various inflammatory mediators have been previously shown to be released into the peritoneal cavity during peritonitis in peritoneal dialysis patients, those that are involved in governing changes in peritoneal permeability to small solutes and protein remain incompletely defined.. We determined the importance of prostanoid production in the enhanced protein loss observed during acute peritonitis by inhibition experiments using indomethacin, an inhibitor of cyclooxygenase activity. The association between changes in peritoneal permeability and the generation of inflammatory mediators after adding Escherichia coli to peritoneal dialysate was first examined in series 1 experiments. Series 2 experiments then determined the effect of intraperitoneal administration of indomethacin (75 microg/mL) on changes in peritoneal permeability after adding E. coli to peritoneal dialysate. All experiments were performed in male New Zealand White rabbits (2.6 to 3.4 kg body weight) using an eight-hour dwell of dialysate containing 2.5% glucose. Peritoneal permeability to creatinine and protein was assessed by time-dependent changes in the dialysate to plasma concentration ratios of these solutes.. Series 1 experiments showed enhanced leukocyte migration into the peritoneal cavity and increased peritoneal permeability to protein during bacterial challenge that was accompanied by an increase in the dialysate concentrations of prostaglandin E2 (PGE2), 6-keto-PGF1alpha, and interleukin-8, but not nitrate + nitrite (a measure of local nitric oxide production). Inhibition of prostanoid production by intraperitoneal administration of indomethacin in series 2 experiments resulted in lower dialysate concentrations of PGE2 and 6-keto-PGF1alpha and in lower peritoneal permeability to protein, both to control levels. No effect of indomethacin on transperitoneal migration of leukocytes or the generation of interleukin-8 was observed.. Enhanced production of prostanoids likely plays an important role in governing the increase in peritoneal permeability to protein during acute, bacterial peritonitis in the rabbit.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Cell Movement; Cyclooxygenase Inhibitors; Dialysis Solutions; Dinoprostone; Escherichia coli Infections; Indomethacin; Injections, Intraperitoneal; Interleukin-8; Leukocytes; Male; Peritoneum; Peritonitis; Prostaglandins; Proteins; Rabbits

The purpose of the study was to characterize oxygen radical generation by emigrated, intraabdominal and circulating polymorphonuclear leukocytes (ePMNLs and cPMNLs) during peritonitis, as well as to assess any differences between oxygen radical production in patients with low Mannheim peritonitis index (MPI < 26, group 1) or high Mannheim peritonitis index (MPI > or = 26, group 2). Lucigenin-enhanced chemiluminescence was used to determine spontaneous and stimulated (FMLP, PMA, and A23 187) oxygen radical generation by ePMNLs and cPMNLs. In group 1 spontaneous and stimulated oxygen radical generation by emigrated PMNLs was markedly enhanced compared to circulating PMNLs (e.g., spontaneous oxygen radical generation: 30.3 +/- 11.8 cpm/cPMNLs versus 107 +/- 46 cpm/ePMNLs, P < 0.05) . In group 2 oxygen radical generation by cPMNLs markedly increased within 48 h after diagnosis of peritonitis and surgery, contrary to radical generation by ePMNLs (e.g., A23 187-stimulated oxygen radical generation 993.7 +/- 350 cpm/cPMNLs versus 285.6 +/- 77 cpm/ePMNLs, P < 0.05. In conclusion, cPMNLs and ePMNLs exhibit marked polymorphism in their capacity to generate oxygen radicals in response to secondary peritonitis. Severe peritonitis (MPI - 26) was associated with a strong increase in oxygen radical generation by cPMNLs without a parallel activity being manifest by ePMNLs.

Topics: Abdomen; Adult; Aged; Case-Control Studies; Cell Survival; Endotoxins; Exudates and Transudates; Female; Free Radicals; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Oxygen; Peritonitis; Prognosis; Tumor Necrosis Factor-alpha

Intra-abdominal infection is one of the major causes of septic shock and multiple organ failure. To date, what causes the disease's progression remains unclear and therefore the relevance of immune modulating therapies remains speculative. The primary outcome measure of this study was to investigate immune modulating mediators at the onset of peritonitis before the development of subsequent septic shock. The secondary outcome measure was to investigate the usefulness of these immune parameters in predicting progression from peritonitis to septic shock. Fifty-eight peritonitis patients were included in this study: 14 patients subsequently developed septic shock. All patients were examined on "diagnosis of peritonitis" (<4 h within establishment of diagnosis), during "early septic shock" (<12 h following the onset of septic shock), and once again during "late septic shock" (within 72-98 h following the onset of septic shock). The immune modulating parameters tumor necrosis factor-alpha (TNF-alpha), the soluble TNF-alpha receptors I and II (sTNF-alpha RI and sTNF-alpha RII), interleukines (IL) -1beta, -6, -8, and -10, and the adhesions molecules endothelial-leukocyte-adhesion-molecule (E-Selectin), intercellular-adhesion-molecule-1 (ICAM-1), and vascular-adhesion-molecule-1 (VCAM-1), in addition to nitrate and nitrite, were determined. In the peritonitis group with subsequent septic shock, TNF-alpha, sTNF-alpha RI + RII IL-1beta, IL-8, IL-10, and nitrate were significantly increased before the onset of septic shock. TNF-alpha had an area under the receiver operating characteristics curve (AUC) of 0.84 and was reliable in predicting the progression from peritonitis to septic shock. The AUC of the other immune modulating parameters, despite being significantly elevated, ranged from 0.71 to 0.76. The AUC of the conventional laboratory markers such as leukocytes and C-reactive protein ranged from 0.64 to 0.68. In peritonitis that progressed to septic shock, an early immune response had already occurred before the onset of septic shock. The progression was best predicted by TNF-alpha. Therefore, mediator therapy might be considered in high-risk peritonitis patients who show an exaggerated immune response before the progression to septic shock.

Topics: Adult; Aged; Aged, 80 and over; Area Under Curve; Disease Progression; Female; Humans; Interleukin-1; Interleukin-10; Interleukin-8; Male; Middle Aged; Peritonitis; Predictive Value of Tests; Receptors, Tumor Necrosis Factor; Shock, Septic; Tumor Necrosis Factor-alpha

Monitoring of peritoneal cytokine concentrations of tumor necrosis factor (TNF)-alpha was recommended for early detection of severe postoperative complications. In the present study the clinical application of cytokine monitoring was examined in the treatment course of severe peritonitis.. Nineteen patients with secondary peritonitis were followed up during 75 abdominal lavages. Serum and peritoneal interleukin (IL)-6, IL-8, and IL-10 and TNF-alpha were measured before the surgical intervention, after 1 hour, 3 hours, 6 hours, and 24 hours. Additionally, cardiorespiratory parameters, osmolarity, C-reactive protein, and total leucocyte count were recorded.. Serum and peritoneal cytokine concentrations did not correlate to each other as well as to the observed cardiorespiratory parameters. Peritoneal cytokine concentrations were 10- to 1000-fold higher to serum concentrations and showed an intermittent wash out. There were no differences in determined cytokine concentrations between survivors and nonsurvivors.. Once elevated, peritoneal cytokine measurements offer no new diagnostic or prognostic tool in abdominal lavage peritonitis treatment.

Topics: Ascitic Fluid; Biomarkers; Blood Pressure; C-Reactive Protein; Cytokines; Female; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Leukocyte Count; Male; Oxygen; Peritoneal Lavage; Peritonitis; Postoperative Complications; Sepsis; Survival Rate; Tumor Necrosis Factor-alpha; Vasoconstrictor Agents

We developed an in vitro model of the peritoneum by coculturing human umbilical vein endothelial cells (HUVEC) and human peritoneal mesothelial cells (HPMC) to gather information on peritoneal physiology and to closer reflect the in vivo situation in humans. HUVEC and HPMC were seeded on collagen-coated polytetraflourethylene-insert membranes of pore size 3 microm. HUVEC were grown on the bottom of the membrane and HMPC on the top. The confluent cells were monitored by measuring transepithelial resistance and by confocal microscopy. The transmigration of PMNs as an important mechanism during secondary peritonitis was studied in this two-chamber model. PMNs were isolated by density separation. After stimulation of HMPC with the complement factor 5 split product C5a (1 ng/ml) or tumor necrosis factor-alpha (TNF-alpha; 10 or 50 microg/ml) for 1 h, 1 x 10(6) PMN were given to the lower compartment. Controls were cocultured cells without stimulation. After 1, 2, and 6 h nonadherent PMNs in the upper compartment were harvested and counted, interleukin-8 was measured in each compartment, and cells on the membrane were paraffin-embedded for immunohistochemistry. Each experiment was performed four times. Cells grew to confluence within 2-5 days and were detected on their respective seeding side by CD34 and cytokeratin 18 counterstaining. Transmigration of PMNs after C5a or TNF-alpha stimulation showed a significant time-dependent increase between 1 h and 6 h (P<0.05). PMNs were found in significantly higher numbers after stimulation with either C5a or TNF-alpha at 1, 2, and 6 h than without stimulation. After stimulation of HPMC, interleukin-8 secretion to the apical compartment increased in a time-dependent fashion, resulting in a gradient between the two chambers. Linear regression analysis revealed significant correlation between transmigrated PMN and interleukin-8 in stimulated cocultures; no correlation was found in controls. This new in vitro peritoneum consisting of cocultured mesothelial and endothelial cells may allow more detailed assessment of peritoneal pathophysiology. Generation of an interleukin-8 gradient affecting the migration of PNMs across the cocultured membrane represents a parameter which may be addressed in further studies.

Topics: Cell Culture Techniques; Cell Movement; Coculture Techniques; Confidence Intervals; Culture Media; Endothelium, Vascular; Epithelial Cells; Humans; Interleukin-8; Linear Models; Microscopy, Confocal; Neutrophils; Peritoneum; Peritonitis; Time Factors; Umbilical Veins

Studies on human macrophages are restricted due to difficulties in isolating significant numbers of human macrophages. High numbers of monocytes/macrophages can be obtained from peritonitis effluents of patients treated with peritoneal dialysis. To determine whether these cells might be useful for functional studies, we characterized peritoneal macrophages (PM) immediately after isolation from the dialysate effluents and their subsequent differentiation. During a 10 days culture period they differentiated morphologically and phenotypically (FACS-analysis) from monocyte-like cells to macrophages. Reflecting the intraperitoneal inflammation we found protein- and mRNA-synthesis of IL-8 and monocyte-chemoattractant-protein-1 (MCP-1) to be upregulated in PM after isolation from the effluents. In contrast, TNF-alpha was downregulated and could not be stimulated by LPS and/or IFN-gamma, reflecting the phenomenon of desensitization. After 10 days in culture, cytokine production normalized to a constitutive level and the TNF-alpha responsiveness to LPS was restored. These data suggest the recovery of PM from the inflammatory prestimulation. Therefore PM harvested from peritoneal dialysis effluents might provide a useful tool for further studies on the role of human macrophages in inflammation.

Topics: Cell Differentiation; Cell Separation; Cells, Cultured; Chemokine CCL2; Humans; Interferon-gamma; Interleukin-8; Lipopolysaccharides; Macrophage Activation; Macrophages, Peritoneal; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Receptors, Cell Surface; RNA, Messenger; Time Factors; Tumor Necrosis Factor-alpha

Leukocyte interactions with vascular endothelium during inflammation depend on cascades of adhesion molecule engagement, particularly during selectin-mediated leukocyte rolling. Leukocyte rolling is also facilitated by members of the integrin and Ig families. Specifically, leukocyte rolling velocities during inflammation are significantly increased in ICAM-1-deficient mice, with ICAM-1 expression required for optimal P- and L-selectin-mediated rolling. Elimination of ICAM-1 expression in L-selectin-deficient mice significantly reduces leukocyte rolling. Whether disrupted leukocyte rolling in L-selectin and ICAM-1 double-deficient (L-selectin/ICAM-1-/-) mice affects leukocyte entry into sites of inflammation in vivo was assessed in the current study by using experimental models of inflammation; thioglycollate-induced peritonitis, chemokine-induced neutrophil migration to the skin, delayed-type hypersensitivity responses, rejection of allogeneic skin grafts, and septic shock. In many cases, the loss of both L-selectin and ICAM-1 expression dramatically reduced leukocyte migration into sites of inflammation beyond what was observed with loss of either receptor alone. In fact, the effects from loss of both L-selectin and ICAM-1 effectively eliminated multiple chronic inflammatory responses in L-selectin/ICAM-1-/- mice. By contrast, the combined loss of L-selectin and ICAM-1 expression had minimal effects on the generation of Ag-specific T cell responses or humoral immunity. Thus, members of the selectin and Ig families function synergistically to mediate optimal leukocyte rolling and entry into tissues, which is essential for the generation of effective inflammatory responses in vivo.

Topics: Animals; Cell Movement; Dermatitis, Contact; Graft Rejection; Immunity, Innate; Immunoglobulin G; Immunoglobulin M; Injections, Intradermal; Injections, Intraperitoneal; Intercellular Adhesion Molecule-1; Interleukin-8; L-Selectin; Lymphocyte Activation; Lymphocytes; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Peritonitis; Shock, Septic; Signal Transduction; Skin Transplantation; T-Lymphocytes; Thioglycolates

Growing evidence supports the idea that adhesion via beta(2) integrins not only allows cellular targeting, but also induces intracellular signaling, which in turn activates functional responses of adherent cells. This study investigates whether beta(2) integrin-mediated adhesion of human polymorphonuclear neutrophils (PMN) has a functional impact on cytokine production. Aggregation of the beta(2) integrin Mac-1 (CD11b/CD18) by antibody cross-linking was found to induce substantial de novo synthesis of IL-8 mRNA as measured by semiquantitative RT-PCR and Northern blotting technique, respectively. Induction of IL-8 mRNA was also observed upon adhesion of PMN to immobilized fibrinogen, a functional equivalent of its clotting product fibrin that serves as a native ligand of Mac-1. Results were confirmed using PMN derived from CD18-deficient mice, which were unable to produce MIP-2 mRNA, a homologue of human IL-8, in the presence of immobilized fibrinogen. In contrast, a substantial increase of MIP-2 mRNA was observed when wild-type PMN were incubated on immobilized fibrinogen. In human PMN, ELISA technique showed that the gene activation that required tyrosine kinase activity resulted in a substantial production and secretion of biologically active IL-8 and IL-1beta. In contrast, no TNF-alpha or IL-6 production was found, revealing that beta(2) integrins mediate differential expression of proinflammatory cytokines. The biological relevance of the present findings was confirmed in an in vivo model of acute inflammation. Altogether, the present findings provide evidence for a functional link between clotting and inflammatory responses that may contribute to the recruitment and/or activation of PMN and other cells at sites of lesion.

Topics: Animals; CD18 Antigens; Cell Adhesion; Chemotaxis, Leukocyte; Cytokines; Fibrinogen; Gene Expression Regulation; HL-60 Cells; Humans; Inflammation; Interleukin-1; Interleukin-8; Macrophage-1 Antigen; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Neutrophil Activation; Neutrophils; Peritonitis; Protein Biosynthesis; Signal Transduction; Transcription, Genetic; Transcriptional Activation

Interleukin-8 and monocyte chemotactic protein-1 (MCP-1) are major leukocyte chemoattractants during bacterial peritonitis by recruiting neutrophils and monocytes/macrophages respectively.. Peritoneal macrophages (PM) from 12 different CAPD patients with peritonitis were stimulated with either 10 ng/ml LPS, 10 ng/ml IFN-gamma or LPS+IFN-gamma, and IL-8 and MCP-1 production was determined on protein and mRNA levels by using ELISA technique and Northern blot analysis. To obtain information from two different stages of activation, experiments were done with highly activated PM directly after isolation and with cells after 10 days in culture, each group being stimulated for 4 h. Unstimulated cells served as control.. Immediately after isolation IL-8 mRNA-expression and synthesis was high and could be further increased by LPS stimulation, whereas IFN-gamma treatment showed no significant influence. The levels of MCP-1 were also initially high but could not be further stimulated by LPS, whereas addition of IFN-gamma resulted in a significant rise in MCP-1 synthesis. After 10 days in culture LPS-stimulation of cells again revealed a significant increase in IL 8 protein synthesis, whereas IFN-gamma showed no effect. LPS anergy for MCP-1 was still seen in PM after 10 days in culture, and IFN-gamma treatment again induced a significant rise in MCP-1 synthesis. The overall production of both chemokines was far higher on day 1 compared to day 10.. Our data show differences in LPS/IFN-gamma regulation for IL-8 and MCP-1 in both highly activated and in resting, mature peritoneal macrophages, suggesting distinct pathways for these chemokines that may offer a means of control for the specific recruitment of neutrophils and monocytes/macrophages in bacterial peritonitis.

Topics: Chemokine CCL2; Gene Expression Regulation; Humans; In Vitro Techniques; Interferon-gamma; Interleukin-8; Lipopolysaccharides; Macrophage Activation; Macrophages, Peritoneal; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Recombinant Proteins; RNA, Messenger

Cytokine levels during infection and sepsis have been extensively studied in the past. In contrast to the excellent data on tumour necrosis factor alpha (TNF-alpha), interleukin 8 (IL-8), and polymorphonuclear (PMN) granulocyte elastase (PMN-E) concentrations in blood, little is known about cytokine and PMN-E levels in tissue or local fluids like abdominal exudate in secondary, purulent peritonitis of man. Therefore, the authors studied perioperative intra-abdominal levels of TNF-alpha, IL-8 and PMN-E in 21 patients with severe purulent peritonitis. The average pre-operative levels of TNF-alpha were 694 +/- 239 pg/ml in exudate and 26 +/- 6 pg/ml in plasma, for IL-8 100 +/- 34 ng/ml and 0.7 +/- 0.5 ng/ml, and for PMN-E 68 +/- 14 microg/ml and 0.7 +/- 0.1 microg/ml, respectively. Standard surgical procedures reduced the intra-abdominal concentrations of cytokines and PMN-E to as low as one tenth of the pre-operative levels. Postoperatively, TNF-alpha and IL-8 levels recovered rapidly and pre-operative levels of IL-8 were reached again after 1 h and for TNF-alpha after 8 h. PMN-E concentration remained below the initial baseline within 8 h of observation. TNF-alpha concentration, but not IL-8 or PMN-E, depended on the microbiological load of the abdominal exudate (< or > 10(3) cfu/ml). There were no significant differences in the intra-abdominal or plasma levels of cytokines or PMN-E between survivors and non-survivors at any observation time.

Topics: Adult; Aged; Aged, 80 and over; APACHE; Exudates and Transudates; Female; Humans; Interleukin-8; Leukocyte Elastase; Male; Middle Aged; Peritonitis; Tumor Necrosis Factor-alpha

An important event in intraperitoneal inflammation is the influx of leukocytes into the peritoneal cavity. Chemokines such as interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) play a major role in the recruitment of immune cells to the site of inflammation. We determined the concentrations of two members of the chemokine family, IL-8 and MCP-1, in the dialysate effluents of 18 continuous ambulatory peritoneal dialysis (CAPD) patients with peritonitis and of 18 non-infected CAPD patients by specific enzyme-linked immunosorbent assays (ELISA). Isolated peritoneal macrophages (PMs) from CAPD peritonitis patients were cultured and IL-8 and MCP-1 production was determined on protein (ELISA) and mRNA level (Northern blot) at designated timepoints over a 72-h culture period. PMs from non-infected patients served as controls. Much higher concentrations of IL-8 and MCP-1 were found in dialysate effluents of peritonitis patients than in effluents of non-infected patients: IL-8 2.39 +/- 1.15 vs 0.05 +/- 0.01 ng/ml and MCP-1 22.5 +/- 6.27 vs 0.42 +/- 0.07 ng/ml. IL-8 and MCP-1 release by cultured PMs from peritonitis patients and non-infected patients revealed significant differences: IL-8 40.3 +/- 2.2 ng/ml after 3 h and 194.2 +/- 34.9 ng/ml after 12 h compared to 21.02 +/- 6.15 ng/ml after 3 h and 89.64 +/- 30.28 ng/ml after 12 h, respectively; MCP-1 3.3 +/- 0.9 ng/ml after 3 h and 25.7 +/- 7.4 ng/ml after 12 h compared to 1.1 +/- 0.2 ng/ml and 1.8 +/- 0.2 ng/ml, respectively. Interestingly, the ratio of IL-8 to MCP-1 concentrations in the dialysate effluents (1:9.4) is reversed in the supernatants of cultured PMs. In the effluents and in the culture supernatants of PMs from CAPD peritonitis patients high amounts of IL-8 and MCP-1 are detectable, suggesting that PMs are an important source for these chemokines during peritonitis. Because of the inverse ratio of IL-8 and MCP-1 in the effluents and culture supernatants it can be assumed that PMs are responsible for the MCP-1 concentration to a lesser extent than for the IL-8 concentration in the effluents.

Topics: Cells, Cultured; Chemokine CCL2; Humans; Interleukin-8; Macrophages, Peritoneal; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; RNA, Messenger

Secretion of IL-2 and sIL-2R in the peritoneal dialysis fluid and in the peripheral blood during peritonitis with reference to the process of IL-6 and IL-8 release were investigated.. 17 patients with end-stage renal disease, treated with continuous ambulatory peritoneal dialysis.. ELISA method using commercial kits, Genzyme Corp Boston and DAKO.. Markedly increased IL-6 (mean; 2895 +/- 1360 pg/ml) and IL-8 concentration (1459 +/- 966 pg/ml) in drain dialysate fluid at the onset of peritonitis, started to drop rapidly in the successive samples after antibiotics had been administered. Statistically significant increase of IL-2 (197 +/- 92 pg/ml) and sIL-2R (287 +/- 79 pg/ml) was observed 16 h later and kept increasing until reaching the peak after 24 h.. Secretion of IL-6 and IL-8 pro-inflammatory cytokines which are mainly synthesized in mesothelium cells is followed by the activation of lymphocytes, their infiltration and the production of T lymphocyte derived IL-2 and sIL-2R.

Topics: Adult; Aged; Ascitic Fluid; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-2; Interleukin-6; Interleukin-8; Kidney Failure, Chronic; Longitudinal Studies; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Receptors, Interleukin-2; Regression Analysis

Interleukin-8 (IL-8) is a chemoattractant that is highly selective for neutrophils. This study was designed to investigate the presence of IL-8 in peritoneal fluid of patients with acute appendicitis. The clinical circumstances underlying the secretion of IL-8 by mesothelium and its mechanism of activation have not been defined. In an in vitro model for bacterial peritonitis the role of bacteria in activating human mesothelial cells to secrete IL-8 was studied. Cultured human mesothelium was incubated with various species of pathogenic bacteria, isolated from peritoneal exudate fluids of patients with appendicitis. The amount of IL-8 secreted by the cultured mesothelial cells was determined in an IL-8 ELISA, as IL-8 was present in the original peritoneal fluid of these patients. Peritoneal fluids from patients with a perforated appendix were found to contain a significantly higher concentration of IL-8 compared to peritoneal fluids from patients with nonperforating appendicitis (121.6 (57.8) ng/ml versus 0.2 (0.07) ng/ml, respectively; mean (SEM), P < or = 0.01). Species of Bacteroïdes and Fusobacterium necrophorum induced IL-8 secretion from cultured mesothelial monolayers to levels comparable to those found in peritoneal fluids in vivo. Heat-killed bacteria and bacterial supernatant were also able to stimulate mesothelium to secrete IL-8. The results suggest that in the early phase of bacterial peritonitis the influx of PMN is regulated by bacteria-induced IL-8 secretion by the mesothelium lining the peritoneal cavity.

Topics: Acute Disease; Appendicitis; Ascitic Fluid; Bacterial Adhesion; Bacterial Infections; Cells, Cultured; Epithelium; Humans; Interleukin-8; Lipopolysaccharides; Peritonitis

Our objective was to determine the incidence of peritonitis episodes with an impaired initial cell reaction (IICR:neutrophil number < 100 x 10(6)/L) over a period of ten years, and to find possible explanations for this unusual presentation of peritonitis. A retrospective review of the files of continuous ambulatory peritoneal dialysis (CAPD) patients included in the CAPD program 1984 and 1993 was done. Analysis of cytokine and prostanoid patterns during four peritonitis episodes with an IICR was compared to 12 episodes with a normal initial cell reaction (NICR). Dialysate cell numbers and immunoeffector characteristics of peritoneal cells were compared in 7 IICR patients in a stable situation and a control group of 70 stable CAPD patients. The setting was a CAPD unit in the Academic Medical Center in Amsterdam. Thirty-five CAPD patients who had one or more peritonitis episodes with an IICR and a control group of 249 CAPD patients were included in the study. The incidence of peritonitis with an IICR was 6%. These episodes occurred more than once in 51% of the patients who presented with IICR. In 72% the cell reaction was only delayed: a cell number exceeding 100 x 10(6)/L was reached later. Staphylococcus aureus was significantly more frequently the causative microorganism compared to all peritonitis episodes (PE) that occurred during the study period. Patients with IICR had lower dialysate cell counts in a stable situation, compared to a control group (p < 0.01). This was caused by a lower number of macrophages and CD4 positive lymphocytes. The phagocytosis capacity of the macrophages appeared to be normal. In a comparison of four PE with an IICR and 12 episodes with an NICR, the tumor necrosis factor-alpha (TNF-alpha) response was similar and occurred on day 1, also pointing to normally functioning macrophages. However, the maximal appearance rates of interleukin-6 (IL-6) and IL-8 occurred later in the episodes with IICR compared to NICR (day 2 vs day 1, p < 0.05). No differences were found in vasodilating prostaglandins, mesothelial cell markers (cancer antigen 125, phospholipids, hyaluronan), and mesothelial cell numbers in the stable situation nor during peritonitis. Peritonitis can present as abdominal pain in the absence of a cloudy dialysate. In some of the patients this presentation occurred more than once. This impaired, most often delayed, cell reaction was associated with a delayed secondary cytokine response. As IL-6 and IL-8 can be synthesiz

Topics: Adolescent; Adult; Aged; Bacterial Infections; Female; Humans; Interleukin-6; Interleukin-8; Kidney Failure, Chronic; Leukocyte Count; Macrophage Activation; Male; Middle Aged; Neutrophils; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Prostaglandins; Retrospective Studies; Staphylococcal Infections; Tumor Necrosis Factor-alpha

Topics: Animals; Blood Platelets; Cell Adhesion; Chemotaxis, Leukocyte; Humans; Immune Complex Diseases; In Vitro Techniques; Interleukin-8; Neutrophils; P-Selectin; Peritonitis; Platelet Membrane Glycoproteins; Rats

Interleukin 8 (IL-8) is one of the most widely studied chemoattractants for leukocytes. It belongs to the newly classified CXC family of chemokine which possesses biological activities mainly on neutrophils. The potential role of IL-8 in inflammation is substantiated by the growing evidences of clinical relevance of IL-8 in various diseases such as infection, ischemia and autoimmune disorders. The common characteristic pathological feature of these events is neutrophil infiltration. Although little is known about the mechanism of neutrophil recruitment into the urine, urinary tract infections (UTI) are accompanied by pyuria. Elevated urinary IL-8 levels were found in patients with UTI. Bioactive, multiple forms of IL-8 were produced locally within the urinary tract, and implied that IL-8 participated in the induction of neutrophil migration into the inflammatory site. Similar findings were observed in the peritoneal dialysate of patients on continuous ambulatory peritoneal dialysis with peritonitis. The notion of involvement of IL-8 in local infection was reinforced by the findings obtained on the rabbit UTI model. Finally, the clinical usefulness, as well as the problems of IL-8 level determination in various body fluids are discussed.

Topics: Humans; Interleukin-8; Peritonitis; Urinary Tract Infections

To evaluate the influence of interleukin-8 (IL-8) and other inflammatory cytokines (IL-6, IL-1 beta and tumour necrosis factor alpha (TNF alpha)) on the occurrence of peritonitis in patients receiving continuous ambulatory peritoneal dialysis (CAPD).. The study population comprised 12 patients with peritonitis, 33 without peritonitis, all undergoing CAPD, and five patients undergoing peritoneal catheter implantation. Cytokine concentrations in dialysis fluid were determined by immunoassay and their values compared.. Concentrations of both IL-8 (median 147 pg/ml, range 20-2273 pg/ml; n = 12) and IL-6 (median 1120 pg/ml, range 96-10,600 pg/ml) were substantially elevated, while the IL-1 beta concentration was lower and TNF alpha was not detectable in patients at diagnosis. The IL-6 concentration was also elevated in patients undergoing catheter implantation as well as in those with peritonitis. The IL-8 concentration, however, was elevated only upon infection. Intraperitoneal production of IL-8 was evident on determination of paired serum and dialysis fluid cytokine concentrations, and immunostaining of peritoneal cells with monoclonal anti-IL-8 antibody.. These results suggest that determination of the IL-8 concentration in dialysis fluid maybe useful as a specific marker for following patients with peritonitis receiving CAPD.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Female; Humans; Immunoassay; Interleukin-1; Interleukin-6; Interleukin-8; Male; Middle Aged; Peritoneal Dialysis; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Tumor Necrosis Factor-alpha

We investigated the role of human mesothelium in an in vitro model of peritonitis with emphasis on the secretion of the neutrophil chemoattractant interleukin-8 (IL-8) and the migration of polymorphonuclear leucocytes (PMN) across monolayers of peritoneal mesothelial cells. PMN showed minimal migration across non-activated mesothelial monolayers (< 2%). However, migration was induced after mesothelial cell activation by IL-1 beta (24%) and this induced migration was significantly blocked by antibodies against IL-8 (63% inhibition; P < or = 0.01). IL-1 beta-activated mesothelial monolayers were shown to secrete IL-8 in a polarized way, which was preferentially oriented towards the apical side of the monolayer. Our results indicate that the influx of PMN into the peritoneal cavity is, at least in part, controlled by the mesothelial cell layer of the peritoneal membrane.

Topics: Cell Movement; Cells, Cultured; Chemotaxis, Leukocyte; Electric Impedance; Epithelial Cells; Epithelium; Humans; Interleukin-1; Interleukin-8; Models, Biological; Neutrophil Activation; Neutrophils; Omentum; Peritonitis

The migration of leukocytes, including polymorphonuclear neutrophils and monocytes, into the peritoneal cavity is a key event of intraperitoneal inflammation. We investigated the levels of two members of the chemokine family, interleukin 8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1), in the dialysate effluent of 18 continuous ambulatory peritoneal dialysis (CAPD) patients with peritonitis and compared them with chemokine levels in noninfected CAPD effluent. Being a major source of inflammatory cytokines, we also isolated peritoneal macrophages from peritonitis effluent to determine the mRNA expression directly after isolation. The mean (SEM) concentrations of IL-8 and MCP-1 were significantly higher in the effluent of peritonitis patients than in noninfected effluents MCP-1: 22.5 +/- (6.27) versus 0.37 +/- (0.1) ng/mL and IL-8: 2.39 +/- (1.15) versus 0.04 +/- (0.01) ng/mL. Northern blot analysis of isolated effluent macrophages revealed strong signals for MCP-1 and IL-8. Our findings showed that CAPD effluent from patients with peritonitis contains markedly elevated MCP-1 and IL-8 levels, suggesting that these chemokines participate in leukocyte recruitment during CAPD peritonitis. Isolation of mRNA of peritonitis-derived peritoneal macrophages revealed strong signals for MCP-1 and IL-8, suggesting that macrophages are a major source of these inflammatory mediators.

Topics: Ascitic Fluid; Blotting, Northern; Chemokine CCL2; Dialysis Solutions; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-8; Macrophages, Peritoneal; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; RNA

CAPD-related peritonitis was used as an in-vivo model to study Il-8 during peritoneal inflammation. Eleven episodes were studied in nine patients, who were followed on 8 consecutive days from the start of peritonitis and once after recovery (control). Il-8 was measured in dialysate (night dwells) and serum. The Il-8 time course was compared to Il-6 and TNF alpha. In addition, an in-vivo relationship between dialysate Il-8 and intraperitoneal accumulation of neutrophils was studied. A highly increased peritoneal appearance rate of Il-8 was found in the acute phase that decreased to control values during recovery. A remarkable parallelism was observed for dialysate Il-8 and Il-6 with respect to the time course and the peritoneal appearance rate. In contrast, the appearance rate of TNF alpha was much less and had a different time course. In three of four cases where the dialysate Il-8 peak occurred on day 2, the dialysate Il-6 peak still coincided with Il-8, in contrast to TNF alpha (always day 1). Dialysate Il-8 generally exceeded serum concentrations during the entire follow-up, indicating intraperitoneal Il-8 synthesis. A positive correlation was present between the dialysate Il-8 peak and the maximal number of neutrophils in dialysate. This relationship was absent for Il-6 and TNF alpha. In five of six episodes where neutrophils were quantified on both day 1 and 2, the Il-8 peak occurred simultaneously with the neutrophil peak. These findings suggest that Il-8 is involved in the recruitment of neutrophils towards the dialysate during peritonitis.

Topics: Acute Disease; Adult; Aged; Ascitic Fluid; Female; Humans; Inflammation; Interleukin-6; Interleukin-8; Kidney Diseases; Leukocytes; Male; Middle Aged; Models, Biological; Neutrophils; Peritoneal Cavity; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Time Factors; Tumor Necrosis Factor-alpha

We investigated interleukin-6 (IL-6) and interleukin-8 (IL-8) in peritoneal dialysate and serum from 17 patients on continuous ambulatory peritoneal dialysis (CAPD) with a total of 24 episodes of peritonitis and from 14 non-infected CAPD controls. Bacterial growth was found in 20 (83%) of the dialysate samples. Staphylococcus epidermidis caused 40% of the culture-verified peritonitis. Samples from dialysate were obtained during the first month of dialysis and during peritonitis from the first three dialysate bags on day 1 (the day of admittance) and from night bags on days 3 and 10. Serum samples were drawn on days 1 and 10. Interleukin-6 was increased in all dialysate samples on day 1. The peak median concentration was 23,500 pg/mL (range 1,710 to 340,000 pg/mL) in the first dialysate. Interleukin-8 was also elevated from all patients in the first dialysate, with a peak median value of 2,000 pg/mL (range, 110 to 185,000 pg/mL). Interleukin-6 and IL-8 concentrations from peritoneal fluid on days 1, 3, and 10 were significantly higher than concentrations from CAPD controls (IL-6 median value, 90 pg/mL, P < 0.001; IL-8 median value, nondetectable, P < 0.001). In serum, IL-6 and IL-8 were detected in 83% and 65% of the episodes, respectively. A correlation (P = 0.007) was seen between IL-6 and IL-8 in the first dialysate sample, but not in the subsequent dialysate samples. The highest acute phase reactant (CRP) level obtained during the peritonitis episode correlated to IL-6 in serum (P < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Aged, 80 and over; Ascitic Fluid; Dialysis Solutions; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Prospective Studies

In this study, we investigated whether peritoneal dialysate interleukin-6 (IL-6) and IL-8 levels were elevated during peritonitis in continuous ambulatory peritoneal dialysis (CAPD) patients, with special reference to the high peritonitis occurrence (HPO) group. Serial measurements of IL-6 and IL-8 levels in dialysate before, during and after resolution of peritonitis were done in 13 CAPD patients with 15 episodes of peritonitis. Based on the peritonitis occurrence, 7 patients were assigned to the low peritonitis occurrence (LPO) and 6 patients to the HPO group. Marked elevation of IL-6 and IL-8 in drain dialysate occurred in the early period of peritonitis especially on the first 2 days in both groups. However, there were no significant differences between the groups in the levels of IL-6 and IL-8 in drain dialysate on the first day of peritonitis. However, the disappearance of peritoneal dialysate IL-8 level was faster in the LPO than in the HPO group. The decrease in IL-8 levels during peritonitis was faster than that of IL-6. Marked elevation of IL-6 and IL-8 in drain dialysate was found in the patient with peritonitis caused by Staphylococcus epidermidis and mixed gram-negative bacilli. Therefore, we hypothesize that when peritonitis occurs too frequently in a short period in the HPO group, more IL-6 and IL-8 have been produced in the peritoneum contributing to the ongoing peritoneal injury and/or fibrosis.

Topics: Adolescent; Adult; Dialysis Solutions; Female; Humans; Interleukin-6; Interleukin-8; Macrophages; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Time Factors; Uremia

An inflammatory reaction was induced in vivo by injection of zymosan into the peritoneal cavity of the rabbit. The inflammatory exudate was found to contain oedema-inducing and neutrophil chemoattractant activity when assayed in rabbit skin in vivo, using 125I-albumin and 111In-neutrophils. This activity was additional to that of complement fragment C5a, which was removed by an affinity gel. Two chemoattractants were isolated by cation-exchange, gel-filtration and reversed-phase h.p.l.c. One of these, which ran as a single band of 6-8 kDa on SDS/PAGE, was subjected to N-terminal sequence analysis without reduction and alkylation of cysteine residues. Positive identification of 28 of the first 31 amino acids revealed a rabbit homologue of interleukin-8 (75% sequence identity with human interleukin-8). The demonstration of interleukin-8 as a major neutrophil chemoattractant in an inflammatory reaction in vivo provides the basis for further investigations into the role of this cytokine in the inflammatory process.

Topics: Amino Acid Sequence; Animals; Chemotactic Factors; Edema; Electrophoresis, Polyacrylamide Gel; Inflammation; Interleukin-8; Molecular Sequence Data; Neutrophils; Peritonitis; Rabbits; Sodium Dodecyl Sulfate; Zymosan