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Intense physical stress, such as that in ultramarathon running, affects the immune system. For monitoring in sports medicine, non-invasive methods, e.g., salivary analysis, are of interest. This pilot cohort study aimed to assess changes in salivary parameters in response to an ultramarathon. The results were compared to blood parameters. Male, healthy finishers (n = 9, mean age: 48 ± 8.8 years, mean height: 1.8 ± 0.1 m, mean weight: 72.5 ± 7.2 kg, mean BMI: 23.5 ± 1.9 kg/cm²) of a 160 km ultramarathon were included. Saliva and blood samples were collected at three time points: T1 (baseline), T2 (shortly after the ultramarathon) and T3 (after recovery). In saliva, cortisol, testosterone, IL-1β, IL-6, IL-8, IL-10, TNF-α, albumin, IgA, α-amylase, aMMP-8, and neopterin were assessed via ELISA. In blood, cortisol, testosterone, IL-1β, IL-6, IL-8, IL-10, TNF-α, blood cell counts, procalcitonin, CRP, osmolality, albumin, and α-amylase were analyzed. The statistical evaluation comprised longitudinal testing and cross-sectional testing between saliva and blood using ratios of T2 and T3 to baseline values. Various parameters in saliva and blood changed in response to the ultramarathon. Comparing blood and saliva, the longitudinal changes of testosterone (p = 0.02) and α-amylase (p = 0.03) differed significantly. Despite the limitations of the study, it underlines that saliva is an interesting option for comprehensive monitoring in sports medicine and necessitates further studies.

Weight gain before the clinical diagnosis of necrotising enterocolitis (NEC) is described as a predictive factor.Weight gain of more than 5% one day prior to clinical suspicion plus increase of plasma Iinterleukin-8 (IL-8) are predictive for NEC.48 infants with diagnosis of NEC stage II and III were enrolled in a case-control study. Oral and parenteral nutrition, diuresis and kinetics of weight and of IL-8 were documented.31 infants with NEC-II and 17 infants with NEC-III were enrolled. Weight gain5% occurred in 35.3% of NEC-III, in 0% of NEC-II and in 4.2% of the control group. IL-8 increased significantly [NEC-III (6 561.4 pg/mL) vs. NEC-II: (326.7 pg/mL) vs. control group (38.9 pg/mL); p0.05]. Sensitivity of IL-8 in NEC-II was 87.10% (70.15-96.25) and in NEC-III 100.00% (80.33-100.00). Sensitivity of weight gain was 0.00% (0.00-11.32) in NEC-II and 35.29% (14.30-61.65) in NEC-III.Weight gain5% was found in only 35.3% of the cases with NEC-III. Combination of weight gain and IL-8 did not improve the diagnosis of NEC.

In der vorliegenden Dissertation wurde die Bakterien-Wirtszell-Interaktion von Bartonella quintana mit humanen THP-1-Makrophagen auf zell- und molekularbiologischer Ebene untersucht. Da das Genom von Bartonella quintana ein Abkömmling des größeren Genoms von Bartonella henselae ist, wurden die Versuchergebnisse im Vergleich zu Bartonella henselae interpretiert.

Infektionen mit Chlamydia trachomatis gehören weltweit zu den häufigsten sexuell übertragbaren Erkrankungen. Sie resultieren in inflammatorischen Erkrankungen wie Zervizitis, Endometritis und Salpingitis. Letztere kann zu ektopen Schwangerschaften und Infertilität führen. Zudem ist C. trachomatis Auslöser des Trachoms, eines der häufigsten Ursachen für Erblindung. All diese Erkrankungen sind charakterisiert durch Entzündungsreaktionen, die zu Gewebeschädigungen, Narbenbildung und Fibrose führen und damit die genannten Krankheiten zur Folge haben. Über die Mechanismen der Interaktion zwischen C. trachomatis und der Wirtszelle ist jedoch noch wenig bekannt. In dieser Arbeit wurde die Rolle der Phosphatidylinositol 3-Kinasen (PI3K), der Mitogen- aktivierten Protein Kinasen (MAPK) und des Transkriptionsfaktors NF-κB an der C. trachomatis- induzierten proinflammatorischen Wirtszellantwort untersucht. Die Infektion von HEp-2 Zellen mit C. trachomatis Serovar K (CTK) führte zu einem zeit- und dosisabhängigen Konzentrationsanstieg des proinflammatorischen Zytokins IL-8 im Zellkulturüberstand. Desweiteren wurde gezeigt, dass die Sekretion von IL-8 offenbar Zelltyp-spezifisch ist, da CTK keinen signifikanten Konzentrationsanstieg von IL-8 im Zellkulturüberstand humaner umbilikalvenöser Endothelzellen (HUVEC) verglichen mit C. pneumoniae TW183 induzierte. Darüber hinaus resultierte die Infektion von HEp-2 Zellen mit CTK in einer dosisabhängigen Aktivierung des Transkriptionsfaktors NF-κB. Interessanterweise waren dazu auch UV-inaktivierte CTK in der Lage. Weiterhin wurde eine Phosphorylierung der MAPKinasen extracellular signal-related kinases (ERK1/2), p38 und c-Jun amino-terminal kinases (JNK) nach Infektion von HEp-2 Zellen mit CTK nachgewiesen. Die nachgewiesene NF-κB- Aktivierung und Translokation in den Zellkern war ebenso wie die konsekutive IL-8- Expression abhängig von der Aktivität der MAPK p38 und ERK1/2 sowie der PI3K, nicht jedoch von der JNK. Diese Beobachtungen haben gezeigt, dass C. trachomatis Serovar K über die Aktivierung verschiedener Signaltransduktionswege in humanen Epithelzellen nicht jedoch in endothelialen Zellen einen proinflammatorischen Phänotyp induziert., Chlamydia trachomatis is the most common cause of sexually transmitted disease. They typically induce asymptomatic lower genital tract infections, such as cervicitis, endometritis and salpingitis. Chronic infections can evolve with sequelae such as ectopic pregnancy and infertility. Moreover, C. trachomatis is the causative agent of trachoma, the leading cause of preventable blindness. Infection is characterized by inflammation which is exacerbated upon reinfection, leading to tissue damage and scarring. Little is known about the mechanisms of C. trachomatis-induced target cell alteration. In this study, we characterized C. trachomatis-mediated effects on host cells. Infection of HEp-2 cells leads to a time and dose-dependent expression of the inflammatory chemokine interleukin-8 (IL-8). While C. trachomatis was also able to infect endothelial cells, it did not induce the expression of endothelial IL-8. These effects were specific for a direct stimulation with viable C. pneumoniae TW183. Furthermore, C. trachomatis induce a dose- dependent expression of the transcription factor nuclear factor-κB (NF-κB) in HEp-2 cells. Interestingly UV-inactivated C. trachomatis had the same effect. We also demonstrated enhanced phosphorylation of the mitogen-activated protein kinase (MAPK) family members extracellular receptor kinase (ERK1/2), p38-MAPK and c-Jun amino-terminal kinase (JNK). Moreover, enhanced activation of the Phosphoinositide 3-kinases (PI3Ks), the MAPKs and the NF-κB was shown. Subsequent IL-8 and NF-κB expression were dependent on PI3Ks, p38-MAPK and ERK1/2 activation as demonstrated by preincubation of HEp-2 cells with specific inhibitors for these pathways. Inhibition of the JNK had almost no effect on IL-8 and NF-κB expression. Thus, C. trachomatis triggers an early signal transduction cascade in target cells that could lead to epithelial but not endothelial cell activation which in turn promote inflammation.

In inhalation therapy, drugs are deposited as aerosols onto the air-facing lung epithelium. The currently used in vitro cell assays for drug testing, however, typically dissolve drugs in the medium, completely covering the cells, which represents an unphysiological drug application scenario. Although physiologically realistic in vitro cell culture models of the pulmonary air-blood barrier are available, reliable, easy-to-handle, and efficient technologies for direct aerosol-to-cell delivery are lacking. Here, we introduce the Air-Liquid Interface (ALI) Cell Exposure-Cloud (ALICE-CLOUD) technology, which uses principles of cloud motion for fast and quantitative delivery of aerosolized liquid drugs to pulmonary cells cultured under realistic ALI conditions. Aerosol-to-cell delivery proved to be highly efficient, reproducible, and rapid when using aerosolized fluorescein as surrogate drug. As a proof-of-concept study for the ALICE-CLOUD, we performed functional efficacy studies with the U.S. Food and Drug Administration-approved proteasome inhibitor, Bortezomib, a novel candidate drug for inhalation therapy. Aerosolized Bortezomib had a pronounced anti-inflammatory effect on human epithelial lung cells (A549), as indicated by a significant reduction of (TNFα-induced) IL-8 promoter activation. Importantly, cell-based therapeutic efficacy of aerosolized Bortezomib under ALI conditions was similar to that under dissolved and nonaerosolized submerged conditions, but with faster uptake kinetics. Our data indicate that the ALICE-CLOUD is a reliable tool for aerosolized drug screening with cells cultured under ALI conditions, which combines ease of handling with rapid, efficient, and dosimetrically accurate drug-to-cell delivery. This may pave the way for screening of inhalable drugs under physiologically more relevant and, hence, potentially more predictive conditions than the currently used submerged cell culture systems.

Die Granulomatose mit Polyangiitis (GPA), ehemals Wegenersche Granulomatose, ist eine komplexe Autoimmunerkrankung der Gruppe der ANCA-assoziierten primären Vaskulitiden. Charakteristisch für die GPA sind chronische nekrotisierende Gefäßwandentzündungen vorwiegend der kleinen bis mittelgroßen Gefäße sowie nicht verkäsende Granulome. In 60-90 % der Fälle ist im Krankheitsverlauf die Schleimhaut von Nase und Nasennebenhöhlen betroffen. Diese stellt somit die häufigste anatomische Lokalisation der GPA-Aktivität dar. Größtenteils ungeklärt ist die Ätiologie der GPA. Als relevanter pathogenetischer Prozess wird eine Dysbalance in der Entzündungsantwort auf mikrobielle Reize diskutiert. Als wichtigstes chemotaktisches Zytokin für neutrophile Granulozyten kommt Interleukin-8 ein besonderer Stellenwert in der Abwehrreaktion gegen mikrobielle Pathogene zu. Interleukin-8 wird von nasalen Epithelzellen in mehrfach höherer Konzentration als alle anderen Zytokine exprimiert und ist darüber hinaus in entscheidender Weise an der Pathogenese der GPA beteiligt. Um die funktionell gestörte nasale Barriere bei GPA-Patienten weitergehend zu untersuchen, wurde in der vorliegenden Studie erstmals das lokale Interleukin-8-Expressionsmusters des Nasenepithels in Bezug auf Prozesse der antimikrobiellen Immunabwehr bei der GPA erforscht. Zunächst erfolgte die Quantifizierung der basalen Interleukin-8-Expression primärer nasaler Epithelzellkulturen von GPA-Patienten im Vergleich zu Normalkontrollen auf Proteinebene. Darüber hinaus wurde untersucht, welchen Einfluss Staphylococcus (S.) aureus als potenter mikrobieller Trigger der GPA-Aktivität auf die Interleukin-8-Sekretion der nasalen Mikroumgebung ausübt. Die Untersuchungen zeigten eine hoch signifikant verminderte basale Interleukin-8-Sekretion des nasalen Epithels bei GPA-Patienten. Ein Zusammenhang zu endonasaler Krankheitsaktivität und immunmodulierender Medikation mit Prednisolon wurde nicht nachgewiesen. Auf Stimulation mit mechanisch lysiertem S. aureus wurde keine quantitative Veränderung der Interleukin-8-Sekretion festgestellt. Die gestörte Interleukin-8-Regulation auf lokaler Ebene kann eine veränderte Zusammensetzung des mikrobiellen Besiedlungsspektrums der Nasenschleimhaut begünstigen, und eine Dysbalance der inflammatorischen Antwort auf mikrobielle Reize fördern. Weitere Untersuchungen sind notwendig, um die pathophysiologische Bedeutung dieser Aspekte besser zu verstehen, und so die Basis für zukünftige therapeutische Ansätze auszubauen.

Mortality of sepsis is still high. Crucial for therapeutic response are the early start of treatment as well as the choice of antibiotics or antibiotic combinations. β-lactam antibiotics with bactericidal mode of action are often recommended in guidelines. But this antibiotic class can trigger the immune system to a maximum by releasing cell wall components or exotoxins. This may lead to a worsening of the patient's clinical situation. In contrast, antibiotics with bacteriostatic action often inhibit bacterial protein synthesis with decrease of production of virulence factors and minimize release of cell wall components. The purpose of this review is to summarise the significance of some bacteriostatic antibiotics and to discuss whether a combination of bactericidal and bacteriostatic agents may improve the course of the illness.

Der LPS-Rezeptor CD14 ist auf BEAS-2B-Bronchialepithelzellen vorhanden und wird zudem als lösliche Form sezerniert. Lipopolysaccharide führen an diesen Zellen zu verstärkter Proliferation, Aktivierung von NFkB und MAP-Kinasen sowie vermehrter Sekretion von Interleukin-8 und RANTES. Die zugrundeliegenden Signalkaskaden beinhalten bei der erhöhten Proliferation p38- und JNK-MAPK, Tyrosin- und PI3-Kinasen, bei der IL-8-Sekretion zusätzlich ERK1/2-MAPK, IRAK, NFkB sowie den IL-8-Rezeptor. Das Bronchialepithel leistet durch die Reaktion auf LPS einen wichtigen Beitrag zur angeborenen Immunität. Die LPS-induzierte Interleukin-8-Sekretion kann durch verschiedene Komponenten von Zingiber officinale (Ingwer) gehemmt werden, sodass diese eventuell für die Behandlung entzündlicher Atemwegserkrankungen in Frage kommen. Asymmetrisches Dimethylarginin hat weder auf die Zytokinsekretion von Bronchialepithelzellen noch auf die Zilienschlagfrequenz oder Kontraktion der Rattentrachea einen Einfluss.

Angiotensin-Converting-Enzym (ACE)-Inhibitoren haben als effektive Antihypertensiva heute einen festen Platz in der Behandlung von Hypertonie und myokardialer Insuffizienz. Seit geraumer Zeit wird auch eine immunmodulatorische Potenz diskutiert. Zur Klärung dieser Fragestellung wurden mittels Vollblut-Assay durchflusszytometrisch die Effekte von ACE-Inhibitoren auf die Funktion neutrophiler Granulozyten und Monozyten untersucht. Anhand eines Kollektivs gesunder Probanden konnten die differenten Effekte von verschiedenen Enalaprilat- und Quinaprilatkonzentrationen auf unterschiedlichen Ebenen der zellulären Immunität zu unterschiedlichen Zeitpunkten determiniert werden. Es konnte gezeigt werden, dass Enalaprilat und Quinaprilat deutliche Effekte auf die schnelle zelluläre Immunantwort besitzten. Dabei wurde die spontane, ebenso wie die Lipopolysacharid (LPS)-induzierte Expression der Oberflächenrezeptoren CR3 (CD11b/CD18) und des LPS-Rezeptors CD14 bereits nach kurzer Inkubationszeit signifikant supprimiert. Außerdem konnte eine signifikante Reduktion in der LPS-induzierten Aktivität der Transkriptionsfaktoren NF-kB und AP-1 sowie eine Hemmung funktioneller Parameter wie Phagozytose und oxidativer Burst dargestellt werden. Zur Ergänzung der durchflusszytometrischen Daten wurden zusätzlich derartige Effekte auf die Synthese des Zytokins Interleukin-8 mittels Sandwich-ELISA untersucht. Zwar zeigen die Ergebnisse ebenfalls eine Reduktion der Interleukin-8-Synthese, auf Grund hoher Standardabweichungen wurde das Signifikanzniveau jedoch nicht erreicht. Die immunmodulatorische Potenz von ACE-Hemmern könnte dazu beitragen, immunassoziierte Erkrankungen positiv zu beeinflussen, oder inflammatorische Risiken operativer Eingriffe zu minimieren. Weitere klinische Untersuchungen z.B. anhand eines kardiochirurgischen Patientenkollektivs sind erforderlich, um diese Ergebnisse hinsichtlich ihrer in-vivo Bedeutung zu klären. Der in dieser Studie verwendete durchflusszytometrische Assay stellt dabei ein schnelles, reproduzierbares Verfahren zu Klärung derartiger Fragestellungen dar., ACE-inhibitors as effective antihypertensive drugs play an important role in the treatment of hypertension and chronic heart failure. Immunmodulatory effects of ACE-inhibitors have been postulated. We therefore evaluated the effects of ACE-inhibitors on neutrophil granulocyte and monocyte function in a whole blood flow cytometric assay. Based on a study population of healthy volunteers we determined concentration and time dependent effects of enalaprilat and quinaprilat on different levels of cellular immunity. We showed a significant inhibition in the LPS-induced expression of complement receptor CR3 (CD11b/CD18) as well as the LPS receptor CD14 after a short incubation time of 15 minutes. We also showed an inhibitory effect of both ACE-inhibitors on LPS-induced activity of transcription factors NF-kB, AP-1 and on functional parameters such as phagocytosis and oxidative burst. Furthermore, the synthesis of the cytokine IL-8, as determined by Sandwich-ELISA, decreased after incubation with ACE-inhibitors. However, this effect failed to be significant because of high standard deviation. ACE-inhibitors could have a positive influence on the therapy of immune changes associated with anaesthesia, trauma or critical illness and could help to decrease the inflammatory complications of surgery. Further clinical investigations, e.g. in cardiac surgery patients, are warranted to verify these results in-vivo. Flow cytometric analyis provides a fast and reproduceable method to answer these questions.

Morbidity and mortality of chronic obstructive pulmonary disease (COPD) are considerable and still increasing. The disease is gaining increasing socioeconomic importance. The knowledge of underlying mechanisms is of special relevance because of the lack of a curative therapy. Respiratory infections have been identified as the most important triggers of acute exacerbations but recent data suggest that they might also play an important role in COPD pathogenesis. This knowledge might offer new therapeutic perspectives in the future. The aim of this review is, therefore, to describe the inflammatory processes involved and to specify the role of respiratory infections in this context.

Chronic obstructive pulmonary disease (COPD) is characterised by a chronic inflammation in the pulmonary tissue. The disease is associated with a switch from a self-limiting inflammatory response, mainly initiated by smoke inhalation, to a chronic persistent inflammatory response after prolonged interaction with cigarette smoke. The development and progression of chronic obstructive pulmonary disease have been associated with increased oxidative stress and reduced antioxidant resources. As a result, the antioxidant capacity decreases in COPD patients. Additionally, there is an imbalance between the release of proteases and of endogenous anti-protease enzymes that prevent elastin digestion. The inflammatory basis of COPD is now well established. In patients with COPD increased numbers of macrophages and neutrophils have been found in induced sputum, the former predominating in mild disease. Biopsy data confirm these observations but also demonstrate increased numbers of CD8-positive T- lymphocytes in the airway wall, which may have a role in the regulation of the inflammatory response to cigarette smoke. The extent of the inflammatory reaction is correlated with the severity of the disease. An imbalance between pro- and anti-inflammatory cytokines may favour this process. Several mediators are involved in COPD including leukotriene B4 (LTB4), tumor necrosis factor alpha (TNF-alpha), and interleukin8 (IL-8). They are selective attractants of neutrophils and regulate the ongoing inflammatory process. The whole process would be predicted to be self-perpetuating leading to a chronic inflammatory state with associated airway remodelling and progressive lung function decline.

Die intestinale Mukosa bildet eine biologisch wichtige Barriere zwischen dem Organismus und den schädigenden Faktoren im intestinalen Lumen. Diese komplexe Aufgabe wird durch eine hochdifferenzierte intestinale Mukosa bewältigt, die eine strukturelle sowie funktionelle intestinale Barriere bildet. Das Ziel der vorliegenden Arbeiten war, ausgewählte Aspekte der Regulations- und Reparaturmechanismen der intestinalen mukosalen Barriere weitergehend zu charakterisieren. Unsere Untersuchungen zeigen, dass das Phospholipid Lysophosphatidsäure (LPA) die intestinale epitheliale Zellmigration stimuliert, die dieser Zellen jedoch inhibiert. Die Modulation der intestinalen Wundheilung durch LPA erfolgt durch einen TGF-b-unabhängigen Mechanismus und wird über einen G-Protein-abhängigen Rezeptor vermittelt, wie wir im Rahmen umfangreicher Untersuchungen zur Signaltransduktion unter Verwendung spezifischer Modulatoren der Signaltransduktion wie Bradykinin, Phorbolester, Pertussistoxin, Suramin und neutralisierender TGF-b-Antikörper belegen konnten. In weiteren Experimenten konnten wir zeigen, dass LPA auch in-vivo einen wundheilungsfördernden Effekt besitzt. Die topische Applikation von LPA in diesem experimentellen Kolitismodell bewirkte einen geringeren Gewichtsverlust sowie ein geringeres Ausmaß an intestinaler Entzündung und Nekrose in-vivo. Diese Untersuchungen legen somit nahe, dass LPA die intestinale epitheliale Wundheilung durch eine Modulation der intestinalen epithelialen Migration und Proliferation durch TGF-b-unabhängige Mechanismen stimuliert. Weitere Untersuchungen beschäftigten sich mit der funktionellen Charakterisierung von Lamina propria T-Zellen (LPT) und peripheren Blut T-Zellen (PBT). Wir konnten zeigen, dass der Zellzyklus von LPT distinkt von PBT reguliert wird. Hierbei spielt der Zellzyklusinhibitor p53 eine zentrale Rolle in der Zellzyklusregulation von LPT. Um Autoimmunität zu verhindern, muss es nach einer Eliminierung des Antigens wieder zu einer Depletion des Pools an Effektor-T-Zellen durch die Aktivierung der Apoptose kommen. Wir konnten zeigen, dass beim Antigen-induzierten Zelltod von LPT der intrinsische Apoptoseweg aktiviert wird und Caspase-8 hierbei eine zentrale Rolle spielt. Physiologischerweise sind Zellzyklus und Apoptose eng miteinander verbunden. In weiteren Versuchen konnten wir jedoch zeigen, dass dies nicht bei LPT der Fall ist und somit die von PBT distinkte Regulation von Zellzyklus und Apoptose mukosaler T-Zellen weiter unterstreichen. Zusammengefasst konnten wir durch ausgewählte Untersuchungen zeigen, dass die intestinale Barriere und ihre funktionelle Beeinflussung eine wesentliche Rolle in der Pathogenese und Therapie intestinaler Entzündungen besitzt. Eine Beeinflussung intestinaler Reparaturprozesse und Modulation abnormer T-Zellen könnte neue Möglichkeiten in der Therapie intestinaler Entzündungen, wie z.B. chronisch entzündlichen Darmerkrankungen bewirken. The intestinal mucosa protect the host from the potential harmful content of the intestinal lumen. To accomplish this difficult goal, the highly complex mucosa forms an anatomical as well as functional barrier to protect the organism.In this work, we aimed to characterize distinct aspect of the intestinal barrier, focussing on distinct regulation and repair mechanism of the intestinal mucosa. First, we demonstrate, that the phospholipid lysophosphatidic acid (LPA) stimulate the migration of intestinal epithelial cells, but, in contrast, inhibit their proliferation. This effect is mediated by G-protein receptors and is TGF-b-independent, as we could demonstrate in further experiments using bradykinine, phorbole ester, pertussis toxin and suramine to modulate distinct signalling pathways.We then demonstrated, using a well-established animal model of colitis, that LPA enhances intestinal wound healing in-vivo. In detail, the topical application of LPA in TNBS-treated rats reduced weight loss, ameliorate intestinal inflammation and prevented necrosis in the animals. This experiments demonstrate for the first time, that LPA modulates migration and proliferation of intestinal epithelial cells by distinct TGF-b independent pathways. Further experiments aimed to explore functional differences between peripheral blood (PBT) and mucosal T-cells (LPT). We demonstrated, that the cell cycle is distinctively regulated in PBT and LPT, identifying p53 as key regulator of LPT cell cycling. To avoid auto-immunity, the pool of effector T-cells must be depleted by apoptosis, once the antigen has been cleared. We demonstrate, that intrinsic pathway of apoptosis is activated during the antigen-induced cell death in LPT and that caspase-8 activity is required to execute LPT apoptosis. Cell cycle and apoptosis are ultimately linked. However, as we show in further experiments, this is not the case in LPT, underlining the distinct regulation of LPT cell cycle and apoptosis.In conclusion, using various distinct experimental tools, we demonstrate that the intestinal barrier itself and the modulation its function plays a fundamental role in the pathogenesis mucosal inflammation. The data presented in this work may therefore open new therapeutic options in the therapy of intestinal inflammatory disorders, such as inflammatory bowel diseases.

The systemic side effects of isolated limb perfusion (ILP) with rhTNF alpha and melphalan are characterised by the induction of a systemic inflammatory response syndrome (SIRS). Procalcitonin (PCT), a serum marker of bacterial sepsis, was investigated with respect to its role in SIRS after TNF-ILP. Serum-PCT was analysed in 24 patients (12 male, 12 female), who treated by ILP for regionally metastasized melanoma (n = 8) or locally advanced soft tissue sarcoma (n = 16). Serum samples were analysed pre- and intraoperatively, and at defined intervals after reperfusion of the limb. In addition to PCT, serum IL-6 and IL-8 were analysed in 11 patients. PCT was significantly elevated over baseline after ILP with a maximum between 8 and 36 hours (p0.001). Even 96 hours after reperfusion, PCT was still significantly elevated as compared to baseline levels (p = 0.005). There was no correlation to the systemic leakage rate during the perfusion. IL-6 and IL-8 were also significantly increased after ILP (p = 0.001), but the maximum peaks of both cytokines were reached much earlier than for PCT (IL-8 max. at 1 hour and IL-6 max. at 4 hours after reperfusion). Serum procalcitonin is induced as part of the specific SIRS after ILP with rhTNF alpha and melphalan. It may be induced directly by rhTNF alpha or by different cytokines, as serum peaks of IL-6 and IL-8 are reached well before the peak of PCT. Determination of PCT prior to and after ILP with TNF might be useful to assess patients at risk of developing hyperdynamic shock.

The appearance of neutrophils in rhinitis and sinusitis led to the working hypothesis that neutrophil-specific attractants commonly called chemokines are generated by stimulation with proinflammatory cytokines and bacteria. The receptor mechanism of chemokine synthesis by bacterial products is under discussion and still has to be elucidated.The primary nasal cultures of epithelial cells and fibroblasts ( n=4) were incubated with TNF-alpha (tumor necrosis factor) for 24 and 72 h. Bacterial stimulation of the cell cultures was performed by adding supernatants from the mucoid phenotype of Pseudomonas aeruginosa (PA01) in dilution 1:5 for 24 and 72 h. Supernatants were collected and the concentration of the chemokines interleukin-8 (IL-8), GRO-alpha (growth-related oncogene-alpha), and ENA-78 (epithelial neutrophil-activating peptide) were determined by ELISA technique. Our results revealed that the protein concentration of the chemokines GRO-alpha and IL-8 was upregulated by TNF-alpha as well as by bacterial supernatants in epithelial cells.We conclude that GRO-alpha and IL-8 were inducible by bacterial supernatants in nasal epithelial cells and fibroblasts.

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), p0.05). Phagocytosis (58 +/- 9 % (ePMNs, peritonitis with complications) versus 81 +/- 6 % (ePMNs, peritonitis without complications) versus 82.2 +/- 1.6 % (ePMNs, controls), p0.05) 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) p0.05) 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.

This study investigated the role of P fimbriae in colonization of Escherichia coli, host response, and bacterial persistence in humans. Human volunteers were inoculated intravesically with the nonadherent ABU isolate E. coli 83972 and with P fimbriated transformants of the same strain. During the following 24 h all urine samples, and thereafter daily samples, were collected for urine culture, analysis of neutrophil numbers, and cytokine concentrations (IL-6 and IL-8). The P fimbriated transformants showed enhanced bacterial colonization in comparison to E. coli 83972 and lowered the bacterial numbers needed for persistent bacteriuria. The P fimbriated transformants also lowered the bacterial numbers needed for a significant neutrophil and cytokine host response. We conclude that P fimbriae enhance bacterial colonization and trigger the host response in the human urinary tract.

Essen, Univ., Diss., 2002 Das 825T-Allel des C825T-Polymorphismus im Gen GNB3, welches für die b-Untereinheit heterotrimerer G-Proteine kodiert, ist mit der Expression der kurzen Spleißvariante Gb3-s und verstärkter Signaltransduktion via Pertussistoxin-sensitiver G-Proteine assoziiert. Da der bg-Untereinheit der G-Proteine eine entscheidende Rolle bei der gerichteten Zellbewegung (Chemotaxis) zukommt, stellte sich die Frage, ob strukturell unterschiedliche G-Proteine, nämlich G-Proteine mit der b-Untereinheit Gb3 und G-Proteine mit der Spleißvariante Gb3-s, unterschiedlich starke chemotaktische Antworten von menschlichen neutrophilen Granulozyten hervorrufen. Menschliche neutrophile Granulozyten bewegen sich gezielt auf Entzündungsherde zu und produzieren zur Bekämpfung von inflammatorischen Noxen Sauerstoffradikale. In dieser Arbeit wurden in Abhängigkeit vom Genotyp am GNB3-Locus (CC-, TC- oder TT-Genotyp) Chemotaxis, intrazellulärer Kalziumanstieg und Superoxid-Generation von menschlichen neutrophilen Granulozyten betrachtet, jeweils nach Stimulation mit den chemotaktisch wirksamen Substanzen N-formyl-methionyl-leucyl-phenylalanin (fMLP) und Interleukin-8 (IL-8). Bei Stimulation mit fMLP wurde eine signifikant gesteigerte Zellmigration bei Neutrophilen von Individuen mit dem TC/TT-Genotyp (EC50 = 1.5 ± 1.3 nM) verglichen mit Neutrophilen von homozygoten C-Allelträgern (EC50 = 5.9 ± 1.5 nM) bei Konzentrationen zwischen 0.1 und 100 nM gefunden. Stimulationen mit IL-8-Konzentrationen zwischen 0.1 und 10 nM zeigten eine bis zu 2.5-fach höhere Chemotaxis der Granulozyten von 825T-Allelträgern im Vergleich zu den homozygoten C-Allelträgern. Im Gegensatz zu diesen Ergebnissen erwiesen sich intrazellulärer Anstieg der Kalziumkonzentration nach Stimulation mit IL-8 und Freisetzung von Sauerstoffradikalen als unabhängig vom Genotyp. Stimulation mit fMLP bewirkte bei T-Allelträgern im Vergleich zu den homozygoten C-Allelträgern eine signifikant kleinere Erhöhung der intrazellulären Kalziumkonzentration. Dies zeigt, daß zelluläre Reaktionen, bei denen Gb3-s beteiligt ist, nicht zwangsläufig erhöht sein müssen. Unter der Annahme, daß eine verstärkte Chemotaxis der Neutrophilen von 825T-Allelträgern gegenüber solchen von homozygoten C825-Allelträgern eine bessere zelluläre Immunkompetenz bedeutet, könnte die Genotypisierung eine wertvolle diagnostische und prognostische Hilfe zur Einschätzung der individuellen Immunkompetenz darstellen.

The healing process of skin wounds is regulated by growth factors which stimulate proliferation of resident cells and their synthesis of extra cellular matrixcomponents. Different leukocyte subtypes (neutrophils, macrophages, lymphocytes and mast cells) participate in wound healing not only as immunological effector cells but also as an important source of inflammatory and growth promoting cytokines. Rapid recruitment of leukocytes and their positioning is tightly regulated by a temporally and spatially changing set of chemokines. Whereas expression of IL-8 and GRO (growth related oncogene) direct early neutrophil recruitment, migration of macrophages is stimulated by MCP-1 (monocyte chemoattractant protein-1) and later, lymphocytes are attracted by IP-10 (gamma-interferon inducible protein-10) and Mig (monokine induced by interferon-gamma). Since chemokines as IL-8 and GRO also stimulate angiogenesis and keratinocyte proliferation, they integrate the inflammatory events with the reparative processes and are potential candidates in the search of wound healing agents.

Peritonitis is characterised by a continued infiltration of the peritoneal cavity with leukocytes, attracted by chemotactic mediators. The aim of this study was to demonstrate the capacity of the human peritoneum to secrete chemokines and to show a therapeutic option by impairing the proinflammatory function of the peritoneum.Peritoneum was obtained from 12 consenting patients undergoing abdominal surgery for noninflammatory diseases. After opening the peritoneal cavity a piece of the parietal peritoneum was excised and subsequently incubated with lipopolysaccharide (LPS, 50 ng/ml) +/- interleukin-10 (IL-10, 100 U/ml) for five hours in vitro. The culture supernatants were assayed for concentrations of interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1) and monocyte inflammatory protein-1 alpha (MIP-1 alpha) by using the ELISA.The cultured peritoneum secreted MCP-1 (mean (SEM): 3416 (659) pg/ml) and IL-8 (2946 (894) pg/ml). The presence of LPS resulted in a fourfold enhancement of this secretion (MCP-1: 13563 (1613), IL-8: 9854 (1305) pg/ml) and led to the production of MIP-1 alpha (1476 (240) pg/ml). The LPS-stimulated production of all of these chemokines was significantly diminished by the presence of IL-10.The reaction of the peritoneum to LPS indicates its proinflammatory function in the context of peritonitis caused by gram-negative bacteria. This inflammatory reaction might be diminished by application of IL-10.

Elective surgical approaches and trauma cause changes in the production of different cytokines, an increased production of acute phase protein and changes in the expression of different cell surface markers.In a prospective study we examined the C-reactive protein level, the production of the cytokines IL-6, IL-8 and IL-1 RA in 25 laparoscopic and 21 conventional cholecystectomies. In addition the cell surface markers CD25 and CD30 on different cell populations and HLA-DR on monocytes were measured. Statistical analysis was made by Student's-t-test and Mann-Whitney's rank sum test.The humoral markers showed a more distinct increase in patients operated on conventionally two and 24 hours after surgery, the differences between the two surgical approaches were significant (p0.01). The cell surface markers CD25 and CD30 showed the same reaction. The HLA-DR expression on monocytes was significantly lower in patients operated on conventionally.Elective surgical approaches cause changes in the immune system, which can be evaluated by the reaction of cytokines and cell surface markers. Laparoscopic cholecystectomies cause less activation of the immune system than conventional operations.

The detection of cytokines may elucidate the pathophysiological mechanisms that produce early systemic complications in acute interstitial (i) or necrotizing (n) pancreatitis (AP). The increase in the level of cytokines in the blood of patients with AP may correlate with the severity of the disease. In a prospective clinical trial from October 1992 to August 1993, 23 patients with AP were recruited and blood samples taken for cytokine detection by commercially available Elisa kits and C-reactive protein (CRP) by laser nephelometry. Six of 11 patients with nAP died either early (n = 1) or of late septic complications. None died of iAP. The peak of cytokine and CRP level in the first 3 days of hospitalization was used for calculation. The IL-6 concentration in the blood reached up to 2600 pg/ml in the 1st few days, depending on the severity of AP, and dropped to almost zero in the next days, independently of the clinical course. The differentiation of i- versus nAP, using a cut-off line of 600 pg/ml, was correct in 20 patients [87%, sensitivity (SE): 82%, specificity (SP): 91%, P0.001]. The blood levels of IL-8 reached a maximum of 1381 pg/ml in the 1st few days, depending on the severity of AP, and showed a correlation with the clinical course in the following days. The peak of IL-8 blood levels indicated correctly the severity of AP in 18 out of 23 patients using a cutoff level of 200 pg/ml (accuracy: 78%, SE: 82%, SP: 75%, P0.01). The CRP levels increased up to a maximum of 535 mg/l and indicated the course of AP correctly in 18 out of 22 patients (SE and SP 82%, P0.01). There was no correlation between cytokine blood levels and mortality. In the blood samples of five patients with i- or nAP, no TNF-alpha was detectable. The blood levels of IL-6, and to a lesser extent of IL-8 and CRP, can predict the severity and early systemic complications of AP. The excessive rise in cytokines can be explained by the stimulation of immunological cells (macrophages, lymphocytes and endothelial cells) in the course of AP, inducing early systemic complications.

We studied the plasma levels of TNF-alpha, IL-6, IL-8 and soluble adhesion molecules (sE-Selectin, sL-Selectin, sVCAM-1) immediately before and during mechanical circulatory support with a Biventricular Assist Device System (BVAD-"Berlin Heart") in comparison to patients with chronic heart failure (NYHA classes II/III) and patients with coronary artery disease with normal ventricular function. Additionally, the biocompatibility of the membranes used in the "Berlin Heart" was tested in vitro. IL-6 and IL-8 but not TNF-alpha could only be detected in patients with cardiogenic shock immediately before starting circulatory support. Furthermore, plasma concentrations of soluble adhesion molecules were statistically significantly elevated in patients with cardiogenic shock compared to patients with coronary artery disease. This picture of a systemic inflammatory response syndrome without significant level of TNF-alpha looks quite similar to that seen in patients following trauma and severe operations. During mechanical circulatory support plasma levels of cytokines and soluble adhesion molecules dropped to low levels in patients, who were successfully maintained on BVAD. By contrast, we have found persistently elevated levels of these mediators in patients with fatal outcome. This seems not to be the result of individual distinct response of blood cells to contact with the artificial surfaces of the device. In summary, our data suggest the development of a systemic inflammatory response syndrome may be due to hypoxia during cardiogenic shock. Persistence of systemic inflammation suggests failing of the mechanical support. Therefore, the monitoring of inflammatory mediators may be relevant as a prognostic marker in these patients (disappearance of peripheral hypoxia).

Thymic peptides show immunoreconstitutive und tumor suppressive effects. They are used in oncology to improve the immunological status of patients. Chemokines are able to activate immune cells and inhibit tumor growth. In this study it was proved whether low molecular thymic peptides are able to increase the secretion of the chemokines monocyte chemotactic protein-1 (MCP-1), interleukin-8 (IL-8), macrophage inflammatory protein-1 alpha (MIP-1 alpha) and macrophage inflammatory protein-1 beta (MIP-1 beta) as well as the cytokine tumor necrosis factor-alpha (TNF-alpha) which is not related to chemokines using cell cultures of human whole blood. The thymic peptides induced a significant (p0.05) elevated secretion of MCP-1 and IL-8 whereas for MIP-1 alpha, MIP-1 beta and TNF-alpha no significant chances were seen. MCP-1 and IL-8 showed divergent dose-dependent effects and a different time kinetic of their secretion. The MCP-1 concentration correlated positively with the count of monocytes in whole blood of the volunteers while the IL-8 concentration in dependence with the incubation time correlated positively with the count of granulocytes or monocytes of the volunteers. The results indicate an activation of monocytes and/or granulocytes by low molecular thymic peptides followed by selective elevated secretion of MCP-1 and IL-8.

Studies of in vivo inhalation of nitrogen dioxide (NO2) have demonstrated a transient pulmonary inflammation. This study was done to determine the contribution of airway epithelial cells to the release of inflammatory mediators following NO2 exposure. Confluent cultures of the human bronchial epithelial cell line BEAS-2B on Transwell-Col filters were exposed for 1 h to air or NO2 up to 1.5 ppm with the apical fluids removed with 5% CO2 at 37 degrees C. The cells were hydrated with Hanks' Balanced Salt Solution (HBSS) in the basolateral compartment. Sequential reverse transcription and quantitative cDNA amplification (RT-PCR) was used to measure inflammatory mediator mRNA abundance in BEAS-2B cultures. When compared to air-exposed cells, NO2 induced increases in IL-6 (23.4-fold) and IL-8 (30.9-fold) mRNA abundance. The NO2-dependent increases in mRNA expression reached a maximum between 0 and 1 h post exposure and returned to baseline levels within 24 h. IL-6 and IL-8 proteins as measured by enzyme-linked immunosorbent assays (ELISA) were also elevated in supernatants recovered from NO2-exposed BEAS-2B cells. These studies suggest that exposure to NO2 induces the synthesis and release of inflammatory mediators from airway epithelial cells that may participate in the pathogenesis of airway disease.

Inflammatory bowel disease (IBD) is characterized by T-cell activation and mucosal influx of inflammatory cells partly mediated by increased local release of cytokines and chemokines. Increased levels of activated platelets are reported in IBD. Activated platelets induce endothelial cells in vitro to secrete several cytokines and growth factors and to express adhesion molecules. This study investigates the expression of interleukin-1 (IL-1), IL-8 and granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors on circulating platelets from patients with IBD and healthy controls and assesses the in vitro effect of various concentrations of IL-1 beta, IL-8 and GM-CSF on platelet activation in healthy controls. Flow cytometry was performed to quantify the percentage of platelets binding phycoerythrin (PE) labeled recombinant human IL-1 beta, IL-8 and GM-CSF. Platelet activation was assessed using fluorochrome labeled anti-GMP-140, an activation-dependent antigen. Results are expressed as percentage cytokine receptor expressing platelets (median and interquartile range IQR). Platelets from patients with IBD expressed significantly more cytokine receptors compared to healthy controls: IL-1R [8.7% (5.5-18.2) vs 3.1% (2.4-4.8), p0.05], IL-8R [22.5% (18.1-27.9) vs 8% (4.5-9.2), p0.001)], GM-CSFR [25.9% (16.1-39.2) vs 3.9% (2.7-3.9), p0.001]. The percentage of activated platelets was significantly increased after in vitro stimulation with IL-1 beta, IL-8 and GM-CSF. We conclude that cytokines and chemokines modulate platelet activation through specific, functional receptors which are upregulated in IBD.

We report on a 45-year-old male patient who presented a classic polyarteritis nodosa (PAN). The clinical course extended over 7 years. In spite of 2 years immunosuppressive therapy with azathioprine and methylprednisolone the course was progressive. Low-dose methotrexate therapy was the only treatment that controlled the disease, leading to rapid clinical and histopathological remission. In low concentrations methotrexate acts as an IL-1 inhibitor, and it obviously suppresses the pathogenetic mechanism of PAN.

Accumulating evidence indicates an association between intraamniotic infection and raising concentrations of amniotic cytokines, resulting in preterm labor and preterm rupture of fetal membranes, because these cytokines are able to stimulate prostaglandin biosynthesis. Therefore the purpose of our study was to investigate if quantitative determination of Il-1 beta, Il-6, Il-8 and TNF-a in amniotic fluid may be a practicable method to diagnose intraamniotic infection. Since invasive amniocentesis doesn't allow repeated cytokine detection, in case of preterm rupture of fetal membranes, amniotic fluid also was obtained by placing a sterile gauze and cotton pad into the women's vagina, absorbing draining amniotic fluid for cytokine detection. Our results clearly indicate that Il-1 beta and TNF-a are not detectable in normal pregnancy, while Il-6 and Il-8 are produced in low, but constant levels. In contrast, in amniotic fluid of patients with intraamniotic infection high amounts of Il-6 and Il-8 were found, while Il-1 beta and TNF-a bioactivity became measurable, indicating that biosynthesis was activated. These results demonstrate, that infection associated cytokines detectable in amniotic fluid are highly sensitive markers for intraamniotic infection. In case of preterm rupture of fetal membranes recovery of amniotic fluid from a vaginal pad allows monitoring of cytokine bioactivity in daily intervals to control success of antibiotic treatment.

IL-8 is a chemotactic cytokine with proinflammatory and growth-promoting activities. Recently it has been shown to influence several functions of keratinocytes, including HLA-DR expression, chemotaxis, and proliferation by binding to a specific receptor. Because psoriasis vulgaris is characterized by epidermal hyperproliferation and infiltration of inflammatory cells, we investigated the expression of IL-8 and its receptor in normal and psoriatic epidermis using semiquantitative reverse-transcriptase-polymerase chain reaction. In addition the mRNA levels of the proto-oncogenes c-ras, c-raf, c-myc, and HER-2 were also investigated as potential growth-promoting stimuli in psoriatic epidermis. IL-8 mRNA was only detected in lesional psoriatic epidermis, and IL-8R-specific mRNA was found to be 10 times increased in lesional psoriatic epidermis. There was no significant difference in the protooncogene mRNA levels. In order to test the relevance of the massively increased IL-8R levels in psoriatic epidermis, we investigated the effect of the antipsoriatic drug FK-506 on specific IL-8 and IL-8R mRNA expression. FK-506 dose dependently inhibited IL-8R expression and function. Our data suggest that in psoriatic skin, elevated IL-8 levels and markedly increased IL-8R expression may act in concert to induce the cardinal signs of psoriasis--epidermal hyperproliferation and leukocyte infiltration. IL-8R may prove a molecular target for antipsoriatic drugs such as FK-506.

NAF/NAP-1 is a novel tissue-derived chemotactic peptide. It consists of 72 amino acids and has no sequence homology to known cytokines. NAF/NAP-1 is produced by a wide variety of cells after stimulation with interleukin-1, tumor necrosis factor or endotoxin, and has the properties of a local mediator of neutrophil recruitment into diseased tissues. There are indications that NAF/NAP-1 is important in the pathophysiology of inflammatory conditions such as psoriasis, idiopathic pulmonary fibrosis, asbestosis, adult respiratory distress syndrome and different forms of arthritis.