Tumor necrosis factor increases serum leptin levels in humans.

Leptin is a pleiotropic hormone believed to regulate body weight. Its function in wasting during inflammatory disease in humans is unknown. We studied the effect of repeated tumor necrosis factor (TNF) infusion on serum leptin levels in six patients with solid tumors. TNF infusion on day 1 resulted in an increase in serum leptin levels from 3.1 (SEM +/- 0.28) ng/mL to 5.2 (SEM +/- 0.6) ng/mL after 12 h (P < 0.001). The serum levels returned to baseline within 24 h. Similar results were obtained when TNF was infused on subsequent days. The study shows that leptin serum levels are under control of TNF.

Clinical relevance of elevated serum thrombomodulin and soluble E-selectin in patients with Wegener's granulomatosis and other systemic vasculitides.

PURPOSE: Vascular injury plays an important pathophysiological role in vasculitis. Soluble serum thrombomodulin (sTM), a promising marker of endothelial cell injury, is released into the circulating blood following cell damage and was therefore investigated as a parameter of disease activity in patients with Wegener's granulomatosis (WG) and various other vasculitides. PATIENTS AND METHODS: One hundred and ninety-seven sera of 102 patients with histologically proven WG of different disease activity and 41 sera of patients with other vasculitides at their active stage were investigated (12 Takayasu arteritis [TA], 7 giant cell arteritis [GCA], 10 polyarteritis nodosa [PAN], 12 Behcet's disease [BD]). The sera were examined for the levels of sTM and sE-selectin (CD62E) by enzyme-linked immunosorbent assay (ELISA) and for the presence of classical anti-neutrophil cytoplasmic antibodies (cANCA) by indirect immunofluorescence (IIF). The disease activity was evaluated according to the clinical symptoms and organ involvement. RESULTS: A significant increase of sTM levels compared with control values (26 +/- 2 ng/ml) was found in active WG, TA, GCA, PAN, and BD: limited active WG: 63 +/- ng/ml; generalized active WG: 119 +/- 15 ng/ml; limited WG, partial remission: 60 +/- 5 ng/ml; generalized WG, partial remission: 75 +/- 7 ng/ml; active TA: 36 +/-; active GCA: 36 +/- 4 ng/ml, active PAN: 33 +/- 2 ng/ml, active BD: 40 +/- 4 ng/ml. Limited and generalized WG in complete remission did not have elevated levels of sTM. sTM values closely reflected relapses and therapy-induced remissions of WG. Elevated cANCA titers were correlated as well with the disease activity in WG but more weakly than sTM levels. In contrast, sE-selectin values were not significantly correlated with the disease activity and the course of disease. CONCLUSIONS: sTM is a promising serological marker of disease activity and progression in active limited and generalized WG and other vasculitides reflecting the degree of endothelial cell damage. sTM might prove to be a clinically useful marker for therapeutic considerations.

Efficient symptomatic control of carcinoid tumors with somatostatin in patients with disease progression under alpha-interferon therapy.

BACKGROUND/AIMS: We report--as a retrospective observation--on eight patients with malignant carcinoid tumors. MATERIALS AND METHODS: All patients were initially treated with alpha-interferon and received the longacting somatostatin analogue octreotide (SMS 201-995) after disease progression. Tumor growth was monitored by CT-scan or ultrasound. In addition, serum CgA and urinary 5-HIAA values were determined. RESULTS: All patients responded with relief of symptoms within a few days after the start of octreotide therapy. A regression of the tumor size did not occur, however four patients showed no significant progress over a period of nine to more than eighteen months. The endocrine parameter chromogranin A--determined by immunoluminometric assay (ILMA)--was elevated in all eight patients regardless of symptoms and showed a close correlation with the course of disease. The urinary 5-HIAA values were only elevated in seven patients. In addition, 123I-SMS 204-090 scintigraphy could be performed in six patients. Using this method most of the primary tumors and metastases could be detected. CONCLUSIONS: Only octreotide therapy showed a sufficient symptomatic control and has to be considered as progress in drug therapy for patients with malignant carcinoid tumors. In addition, chromogranin A is an interesting endocrine parameter for the follow-up of the secretory activity.

[Respiratory symptoms and sensitization to airborne pollen of ragweed and mugwort of adults in Southwest Germany]. / Atemwegsbeschwerden und Sensibilisierungen gegen Pollen von Ambrosia artemisiifolia und Beifuß bei Erwachsenen in Süd-West-Deutschland.

BACKGROUND AND OBJECTIVE: Allergic diseases of the respiratory tract are common atopic diseases in the population. Pollen of plants are one of their main causes. Pollen of trees, grasses, and weeds like mugwort are of particular importance in this context. The purpose of the present study was to determine the association between typical respiratory symptoms due to pollen allergens and the sensitization to pollen of ragweed and mugwort in adults. METHODS: 1039 participants (18 to 66 years) from Southwest Germany were included in this cross-sectional study. Complains typically for aeroallergens were recorded by questionnaire. In-vitro existing sensitizations were determined for grasses/early bloomer (gx1), trees (tx6), mugwort (w6 and Art-v1) and ragweed (w1 and Amb-a1) by testing for specific IgE-antibodies. RESULTS: In a screening test with a mixed allergen sample (sx1) 36.0 % of the participants showed an aeroallergen sensitization. Consecutive investigations of these 374 positive samples revealed a sensitization to grasses/early bloomer, trees, ragweed (w1), mugwort (w6) and the major allergens ragweed (Amb-a1) and mugwort (Art-v1) in 61.8 %, 54.5 %, 29.9 %, 24.3 %, 1.9 %, and 12.3 %, respectively. This corresponds to 22.2 %, 19.6 %, 10.8 %, 8.7 %, 0.7 % and 4.4 %, respectively, in the whole study population. The participants tested positively stated significantly more disorders. On average, 51.1 % had known hay fever, 65.2 % sneezing without cold and 41.5 % sneezing due to contact with grasses or flowers. In contrast, participants tested negatively stated disorders in only 4.4 %, 32.5 %, and 3.9 %, respectively. CONCLUSION: In Southwest Germany, sensitizations to plant pollen mainly are still caused by grasses, trees, and weeds like mugwort. A sensitization to pollen of ragweed alone is rare until now. However, in case of continuous allergic disorders of the respiratory tract in late summer, a sensitization to ragweed can be important for differential diagnostics.

[Sensitization to airborne ragweed pollen--a cause of allergic respiratory diseases in Germany?]. / Sensibilisierung gegen Ambrosiapollen--Eine Ursache für allergische Atemwegserkrankungen in Deutschland?

BACKGROUND AND OBJECTIVE: Allergic skin and respiratory diseases show a high prevalence in most industrial countries. In addition, during the last years ragweed colonization has increased in Europe. Ambrosia pollen ( AMBROSIA ARTEMISIIFOLIA L. - common ragweed) are highly allergenic. Due to the late flowering time (august/September) of ragweed this can result in increasing health threats for allergic populations. This is of particular importance for those who already are sensitive to some grass or tree pollen. These individuals can then suffer from allergies during nearly the whole year. The present study examined the prevalence of sensitization to ragweed in German children and possible health implications. METHODS: Between 2004 and 2007 sera of 1323 10-years old children in Baden Württemberg were tested in-vitro for specific IgE-antibodies against common aeroallergens including ragweed pollen. RESULTS: Specific IgE-antibodies to extracts of common ragweed pollen were present in 10 - 17 % of the tested sera depending on the year of investigation. CONCLUSION: The determined specific IgE-antibodies may be the result of a direct sensitization to ragweed pollen or correspond to cross-reactivity to other plants of the asteraceae subfamily or some nutritional allergens. The detection of sensitization to ragweed pollen does not prove actual allergic disease. However, a ragweed derived allergy should be considered in the differential diagnosis when allergic symptoms are present in direct connection to the flowering-time of ragweed. Ragweed plants should be removed and the spread of the plant 'restricted', as experiences in other countries with already wide spreading show.

[Present importance of direct immunologically based intervention strategies using anticytokines in rheumatoid arthritis]. / Gegenwärtige Bedeutung der direkten immunologischen Interventionstherapie durch Antizytokine bei der Rheumatoiden Arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory multisystemic autoimmune disease of unknown origin. RA is clinically characterized by recurrent inflammation of joints, synovialitis, progressive destruction of cartilage or bone tissue and multiorgan involvement. Today all established therapies of RA are still unable to stop or even cure the disease. In most cases these therapies can only reduce progression. Furthermore, these therapies have substantial side effects, which can contribute to the increased mortality of disease. Therefore more effective therapies with fewer side effects are needed. In this context direct immunological intervention strategies increasingly gained interest to inhibit proinflammatory cytokines. In vivo and in vitro studies as well as experimental therapies documented the important role of the proinflammatory cytokines TNFalpha and IL-1 in RA. The therapy with TNFalpha-antibodies or receptor fusion proteins as well as IL-1 receptor antagonists proved to be clinically as well as immunologically highly effective as therapy of RA. The single dose treatment is associated with mild side effects only. In addition, trials using combined TNFalpha-antibody and methotrexate therapy gave promising results. However, potential severe side effects may occur after repeated therapy cycles or may be discovered after prolonged time of observation only (e.g., allergic reactions, induction of autoantibodies or malignancies). Therefore, at present these therapy options can only be recommended for selected patients, who are included into controlled clinical trials. In addition, repeated courses of therapy seem to lead to reduced therapeutical efficacy (especially in TNFalpha-antibody therapy). Further controlled studies with cytokine antagonists should especially address these problems and focus in particular on potential inductions of autoantibodies or malignancies as well as on additional long-term side effects. In contrast to direct inhibition of TNFalpha or IL-1 several further therapies indirectly influence these cytokines by interference with their synthesis or by alteration of the respective receptors. The importance of these therapeutical options has to be determined as well as the possibility of combination of established therapies with immunological intervention strategies.

Autoantibodies to intermediate filaments in sera of patients with Schistosoma mansoni infection.

Autoantibodies to the intermediate filament proteins vimentin and keratin were studied in sera of 50 Caribbean patients with Schistosoma mansoni infection and 50 control subjects. Autoantibodies were detected by indirect immunofluorescence on HEp-2 cells pretreated with colchicine. The incidence of anti-vimentin antibodies in patients' sera was 94% for IgM, 12% for IgG, and 4% for IgA; in the control subjects incidence was 52%, 0%, and 4%, respectively. Anti-keratin antibodies were found in 82%, 4%, and 4% of patients' sera and 42%, 0%, and 2% in controls, respectively. The difference between the geometric means of titres for patients (1:150) and controls (1:26) was highly significant (P less than 0.001). The possible role and genesis of autoantibodies to intermediate filaments is discussed.

Tietze's syndrome--a chameleon under the thoracic abdominal pain syndrome.

Tietze's syndrome is characterized by pain due to a self-limiting, localized, non-suppurative swelling of the costochondral or sternoclavicular junction of unknown etiology. The case of a woman is presented here who was submitted to the hospital under the suspicion of a pathological fracture. After extensive investigations the diagnosis of Tietze's syndrome was made by exclusion, and the patient was successfully treated with local injection of an anaesthetic. Possible differential diagnoses of Tietze's syndrome include myocardial infarction, pneumonia, and others. This report emphasizes the importance of a thorough clinical investigation and the need for the exclusion of severe and lifethreatening diseases.

Serum levels of adhesion molecules and thrombomodulin as indicators of vascular injury in severe Plasmodium falciparum malaria.

Severe Plasmodium falciparum malaria is characterized by multiple organ involvement due to sequestration of infected erythrocytes in small vessels. Endothelial cell adhesion molecules play an important role in this interaction. During the course of a severe cerebral P. falciparum malaria infection we found very markedly elevated levels of the soluble adhesion molecules intercellular adhesion molecule-1, E-selectin, and vascular cell adhesion molecule-1, with a maximum increase of nine, seven, and eight times, respectively. These very high levels of soluble adhesion molecules point to an endothelial cell injury as an additional cause to physiological release or shedding due to receptor interactions. Soluble thrombomodulin (sTM) levels showed an extremely marked elevation up to 332 ng/ml (up to 13 times the normal value) as well. Malaria patients without severe organ involvement/cerebral manifestation showed only a mild elevation of sTM levels. TM is a parameter independent of the immunological system. It is regarded as a marker of vasculitis and endothelial cell destruction. Therefore, markedly elevated sTM levels document a substantial endothelial cell injury in severe malarial infection and may be of diagnostic and prognostic importance.

Extrapulmonary sarcoidosis primarily diagnosed in the liver.

Sarcoidosis is a relatively common, chronic, multisystem disease of unknown origin characterized by the presence of noncaseating epithelioid granulomas. Although an array of organs may be affected by the disease, the commonest site of affection is the lung. We describe a 73-year-old patient admitted to our hospital because of fatigue, weight loss, and an increased alkaline phosphatase level. In conjunction with clinical presentation, laboratory variables, and imaging analysis, a liver biopsy finally confirmed the diagnosis of a systemic sarcoidosis without affection of the lung or mediastinal lymph nodes. Treatment with ursodeoxycholic acid before diagnosis did not improve clinical symptoms and cholestasis indicators. After prednisone treatment, liver enzyme values normalized and remained normal during follow-up for 2 years after diagnosis. The literature on hepatic manifestation of sarcoidosis, its diagnosis, treatment, and prognosis is reviewed. This single case of sarcoidosis presented to the clinician almost exclusively with liver enzyme abnormalities. The consideration of sarcoidosis in such cases is of utmost importance, since the differential diagnosis of hepatic granulomas includes infectious diseases in which treatment with corticosteroids could be fatal.

Intraoperative endothelial damage is associated with increased risk of stenoses in infrainguinal vein grafts.

OBJECTIVES: Saphenous vein injury is believed to predispose to vein graft stenoses. The aims of this study were to assess the endothelial injury associated with infrainguinal vein bypass surgery by serial measurements of serum thrombomodulin concentration as well as by platelet scintigraphy, and to relate these findings with the postoperative development of stenoses. METHODS: In 35 patients undergoing vein bypass surgery serum thrombomodulin concentration was measured pre- and postoperatively. Autologous 111-indium labelled platelets were administered into the common femoral artery immediately after restoration of flow in the graft and scintigraphic images were obtained 4 and/or 24 h later. RESULTS: Serum thrombomodulin increased markedly from median 17 ng/ml preoperatively to 32 ng/ml 1 day after surgery (p =0.00002). Platelet scintigraphy revealed a total of 62 focal activity accumulations, the majority being located in the anastomotic regions. Among the 30 patients with grafts remaining patent at 30 days stenoses occurred in nine (16%) of 55 regions with scintigraphic platelet accumulations as compared to only four (4%) of 94 regions without platelet accumulations (p =0.03). CONCLUSIONS: The very high predictive value of a negative platelet scintigraphy (96%) strongly suggests that localised perioperative endothelial injury is an important pathogenetic factor in the development of vein graft stenoses.

Serum thrombomodulin-a reliable marker of disease activity in systemic lupus erythematosus (SLE): advantage over established serological parameters to indicate disease activity.

To date no specific serological parameter is available to assess disease activity in SLE. Soluble serum thrombomodulin is a new marker of endothelial cell injury and vasculitis. The objective of this study was to compare in vivo soluble thrombomodulin as marker of disease activity in SLE with established and recent serological parameters. One hundred and twenty-four sera of 30 patients with proven SLE with different disease activities were tested for serum levels of thrombomodulin, intercellular adhesion molecule-1 (ICAM-1), E-selectin, vascular cell adhesion molecule-1 (VCAM-1), IL-2R, IL-6, IL-10, dsDNA by ELISA and dsDNA additionally by radioimmunoassay (RIA). C-reactive protein (CRP), complement component C3, IgG, creatinine, anti-nuclear antibodies (ANA) and intermediate filament antibodies were measured by standard laboratory tests. The clinical disease activity was evaluated by the Systemic Lupus Activity Measure (SLAM). Correlations of the different serological SLE disease activity parameters with the SLAM scores revealed the highest significance for serum thrombomodulin (correlation coefficient 0.82). This was further confirmed by the intra-individual analysis of follow-up sera. In addition, a moderate correlation could be found for IL-6, IL-10, ICAM-1, CRP and erythrocyte sedimentation rate (ESR). In summary, soluble thrombomodulin is the most important serological parameter of disease activity in SLE currently available, as shown by the in vivo studies. Soluble thrombomodulin might be a valuable serological parameter for therapeutical considerations.

Interaction of endothelial cells and neutrophils in vitro: kinetics of thrombomodulin, intercellular adhesion molecule-1 (ICAM-1), E-selectin, and vascular cell adhesion molecule-1 (VCAM-1): implications for the relevance as serological disease activity markers in vasculitides.

Recently markers of endothelial cell activation or injury gained increasing interest as serological parameters of disease activation in vasculitides. Among these, soluble serum thrombomodulin, ICAM-1, VCAM-1 and E-selectin are of particular interest. However, only thrombomodulin showed the expected close correlation. The objective of this study was to investigate in vitro the kinetics of these endothelial cell receptors after interaction of unstimulated or cytokine-activated polymorphonuclear neutrophils (PMN) and endothelial cells in order to find evidence explaining these different clinical findings. Over the time period of up to 48 h of incubation the kinetics of thrombomodulin, ICAM-1, E-selectin, and VCAM-1 levels in the supernatant of endothelial cells in co-culture with neutrophils were determined in vitro by ELISA under basal and partially cytokine-activated (tumour necrosis factor-alpha) conditions. Increased levels of ICAM-1, E-selectin and VCAM-1 were already found due to cytokine activation of endothelial cells alone. This increase was augmented after coincubation with neutrophils. In contrast, a significant increase of thrombomodulin in the supernatant was only found due to cell injury after cell-cell interaction of cytokine-activated endothelial cells with neutrophils. In conclusion, this in vitro model of the kinetics of soluble endothelial cell receptors after cell-cell interaction of cytokine-activated PMN and endothelial cells underlines the advantage of thrombomodulin in contrast to the adhesion molecules as a marker of endothelial damage. Therefore, soluble thrombomodulin seems to be a promising, valuable serological disease activity marker in vasculitides.

Elevated serum levels and reduced immunohistochemical expression of thrombomodulin in active ulcerative colitis.

BACKGROUND & AIMS: The pathogenesis of ulcerative colitis and Crohn's disease is still unclear. Vascular injury has been suggested as a potential pathogenetic mechanism. Serum thrombomodulin is a marker of endothelial cell injury. The aim of this study was to determine the relevance of increased serum thrombomodulin levels for assessing disease activity in inflammatory bowel disease. As a potential cause of serum thrombomodulin level increase, the loss of local vascular thrombomodulin expression was investigated immunohistochemically. METHODS: Thrombomodulin levels were determined by enzyme-linked immunosorbent assay in sera from patients with ulcerative colitis, Crohn's disease, Schistosoma mansoni infection, and infectious diarrhea and controls. The vascular expression of thrombomodulin was investigated immunohistochemically in fresh frozen transmural specimens of normal, Crohn's, and ulcerative colitis bowel samples. RESULTS: Significantly elevated serum thrombomodulin levels were only detected in active ulcerative colitis and infectious diarrhea complicated by septicemia. A marked and general loss of vascular endothelial cell thrombomodulin expression was found immunohistochemically in inflamed bowel tissues. Graded by a newly established thrombomodulin staining index, this was significantly more marked in ulcerative colitis than Crohn's disease. CONCLUSIONS: Serum thrombomodulin proved to be a novel marker of disease activity in ulcerative colitis closely related to local vascular endothelial cell damage, which might be a relevant pathophysiological feature of ulcerative colitis.

Release of thrombomodulin from endothelial cells by concerted action of TNF-alpha and neutrophils: in vivo and in vitro studies.

Inflammatory cytokines decrease the expression of thrombomodulin (TM) on the endothelial cell surface by suppression of TM transcription and translation or internalization with subsequent degradation. Nevertheless, elevated serum TM levels are found in diseases associated with systemical or locally increased levels of inflammatory cytokines. To study directly the in vivo effects of tumour necrosis factor-alpha (TNF-alpha) we determined the course of serum TM after systemic recombinant human (rh)TNF-alpha therapy. The TM levels were determined by enzyme-linked immunosorbent assay (ELISA). Systemic rhTNF-alpha therapy resulted in a marked and significant increase of serum TM. Using a mouse model we studied whether increased serum TM is associated with a decreased expression of TM on the endothelial surface in vivo. The immunohistochemical staining of the vasculature of meth-A sarcoma transplanted in mice showed a loss of TM immunoreactivity 4 hr after intravenous TNF-alpha application. To study the mechanism of TNF-alpha mediated release of TM, cultured endothelial cells were incubated with neutrophils and TNF-alpha. Incubation with TNF-alpha alone did not lead to an increase of TM in vitro. However TM was released into the culture supernatant when endothelial cells pretreated with TNF-alpha were exposed to neutrophils. This was associated with morphological evidence of endothelial cell damage. Therefore, the concerted action of cytokine-stimulated endothelial cells and neutrophils results in release of TM from cultured endothelial cells after rhTNF-alpha therapy. This might explain the increased serum TM levels observed in diseases associated with increased systemic or local levels of inflammatory cytokines despite the induced internalization and the direct inhibitory effects of TNF-alpha on TM transcription and translation.

Kinetics of soluble TNF-receptors and soluble adhesion molecules ICAM-1, E-selectin and VCAM-1 under systemic rhTNF alpha therapy.

Cytostatic as well as cytotoxic effects of tumour necrosis factor alpha (TNF-alpha) therapy have been shown in vitro and in experimental in vivo models. Nevertheless, the mechanism of anti-tumour activity in humans in vivo remains unclear. To determine the role of the vascular lining endothelial cells as important mediators of several immunological interactions, we investigated changes in the levels of the soluble endothelial cell adhesion molecules intercellular adhesion molecule 1, E-selectin and vascular cell adhesion molecule 1 as well as of soluble TNF receptors I and II during systemic therapy with recombinant human rhTNF-alpha (rhTNF-alpha). All tests were performed by enzyme-linked immunosorbent assays (ELISAs). The clinical efficacy of the intravenous rhTNF-alpha therapy was poor. Only one patient with isolated intraarterial limb perfusion had a delayed, marked, but only temporary necrosis of tumour cells. In contrast, we found a marked, significant and (during therapy) undulating augmented increase in the levels of soluble adhesion molecules as well as of the soluble TNF receptors. Taken together, these data support the hypothesis that a sufficient tumour-specific cellular immunity is required to achieve a clinically apparent efficacy of systemic rhTNF-alpha therapy in addition to cytokine-dependent inducible activation mechanisms. In this context, the vascular lining endothelial cells might play an important role as mediators of the complex immunological antitumoral activity.

Serum thrombomodulin. A novel marker of disease activity in systemic lupus erythematosus.

OBJECTIVE: Soluble thrombomodulin (sTM), a proposed serum marker of endothelial cell injury, was investigated as a parameter of disease activity in patients with systemic lupus erythematosus (SLE). METHODS: Levels of sTM were determined by enzyme-linked immunosorbent assay. Disease activity was assessed using 3 established scoring systems: the American College of Rheumatology (ACR), the New York Hospital for Special Surgery (NYHSS), and the Systemic Lupus Activity Measure (SLAM) systems. RESULTS: A close correlation was found between sTM levels and disease activity as assessed with all 3 scoring systems: r = 0.52 by the ACR, 0.75 by the NYHSS, and 0.82 by the SLAM. CONCLUSION: We found that sTM is a sensitive serologic marker of organ involvement in patients with SLE. Furthermore, sTM may prove to be an important marker for vasculitis in general.