Modulatory effect of sera from patients with various types of pulmonary fibrosis on mononuclear cell induced angiogenesis in relation to pulmonary function.

Angiogenesis plays an important role in the pathogenesis of idiopathic pulmonary fibrosis. Pulmonary fibrosis occurs also in many diseases, such as other types of interstitial pneumonias or drug-induced pulmonary fibrosis. The aim of the study was to examine the effect of sera from patients with various types of pulmonary fibrosis on angiogenesis induced by human mononuclear cells (MNC) in relation to lung functions. The study population consisted of 32 patients with idiopathic pulmonary fibrosis (IPF), 11 patients with drug-induced pulmonary fibrosis (DIPF), 6 with cryptogenic organizing pneumonia (COP), and 20 healthy volunteers. An animal model of leukocyte-induced angiogenesis assay was used as an angiogenic test. Spirometry, whole-body plethysmography, static lung compliance (Cst), and diffusing capacity of the lung for CO (DL(CO)) were performed in all patients. Sera from IPF and COP patients significantly stimulated angiogenic activity of MNC, compared with sera from healthy donors and from DIPF patients (P<0.001). However, sera from healthy donors and DIPF significantly stimulated angiogenic activity of MNC compared with the control group with PBS (P<0.001). In all groups, a decrease in the mean value of Cst and DL(CO) was observed, but no significant correlation between VC, FEV(1), DL(CO), Cst, and angiogenic activity of sera from examined patients was found. Sera obtained from patients with pulmonary fibrosis constitute a source of mediators modulating angiogenesis, but the pattern of reaction is different in various diseases. The strongest reaction is observed in IPF and the weakest one in DIPF. The angiogenic activity of sera did not correlate with the pulmonary function of patients with pulmonary fibrosis.

Angiogenic activity of sera from silicosis and pulmonary Langerhans cell histiocytosis patients in relation to lung function tests.

Angiogenesis has been implicated in the pathogenesis of interstitial lung diseases. A correlation between serum angiogenic cytokines level of patients with idiopathic pulmonary fibrosis and radiographic manifestations or functional pulmonary changes has been described, but the role of angiogenesis in the pathogenesis of other interstitial lung diseases such as silicosis and pulmonary Langerhans cell histiocytosis remains unclear. The aim of the study was to examine the effect of sera from silicosis and pulmonary Langerhans cell histiocytosis patients on angiogenesis induced by human mononuclear cells (MNC) in relation to pulmonary function. The study population consisted of 12 patients with silicosis, 12 patients with pulmonary Langerhans cell histiocytosis (PLH), and 14 healthy volunteers. Spirometry, whole-body plethysmography, static lung compliance (Cst), and diffusing capacity of the lung for CO (DL(CO)) were performed in all patients. As an angiogenic test, leukocyte induced angiogenesis assay according to Sidky and Auerbach was used. Sera from PLH patients exerted a significant inhibitory effect on angiogenesis (P<0.001). Sera from silicosis patients significantly (P<0.001) stimulated angiogenesis compared with sera from healthy donors. However, sera from healthy donors significantly stimulated the angiogenic activity of MNC compared with the control with PBS. The mean value of DL(CO) was significantly lower in the group of patients with PLH compared with patients with silicosis (P<0.05). A significant correlation between angiogenesis index and DL(CO) was observed (P<0.05). No significant correlation between the angiogenesis index and other functional parameters was found. Sera from interstitial lung diseases patients and healthy donors constitute a source of mediators modulating angiogenesis. Sera from silicosis patients stimulate neovascularization but sera from PLH patients exert an inhibitory effect on angiogenesis. A correlation between serum angiogenic activity and DL(CO) was found.

Angiogenic activity of sera from patients with systemic autoimmune diseases in relation to clinical, radiological, and functional pulmonary status.

Systemic autoimmune diseases, such as vasculitis and collagen diseases, are characterized by chronic inflammation. Mutual interrelationship between angiogenesis and chronic inflammation has already been demonstrated. The aim of the study was to examine the effect of sera from patients with systemic autoimmune diseases on angiogenesis induced by human mononuclear cells. The study population consisted of 43 patients with a systemic autoimmune disease associated with pulmonary manifestations, divided into three groups: 14 with Wegener's granulomatosis (WG), 13 with systemic sclerosis (SS), and 16 with collagen vascular diseases (CVD) such as rheumatoid arthritis, systemic lupus erythematosus, and dermatomyositis. The control group consisted of 15 healthy volunteers. Clinical status was evaluated using a questionnaire. Standard chest radiographs were performed in all patients. Pulmonary function tests were performed according to the ERS standards. An animal model of a leukocyte-induced angiogenesis assay was used as an angiogenic test. Sera from WG and CVD patients significantly stimulated angiogenesis compared with healthy subjects (P<0.001). On the other hand, sera from healthy donors exerted a proangiogenic effect compared with PBS. In contrast, sera from SS patients significantly (P<0.001) inhibited angiogenesis compared with sera from healthy subjects and PBS. Proangiogenic effect of sera from systemic diseases patients depended on radiological changes. No significant correlation between a degree of dyspnea or functional pulmonary tests and the number of new vessels or angiogenesis index was found. Sera from patients with systemic autoimmune diseases and healthy people constitute the source of mediators modulating angiogenesis. These modulatory effects differ depending on the disease entity.

TNFalpha and INFgamma inducing capacity of sera from patients with interstitial lung disease in relation to its angiogenesis activity.

Sera from interstitial lung diseases (ILD) constitute a source of mediators participating in angiogenesis. The nature of these mediators is unknown. The aim of our study was to asses whether preincubation with sera from ILD patients could influence TNFalpha and INFgamma production by normal mononuclear cells (MNC) challenged with LPS (for TNFalpha) or PHA (for INFgamma), and to correlate the cytokine levels with angiogenic properties of sera. The study population consisted of 53 patients with ILD, 16 with sarcoidosis (SAR), 11 with avian fanciers' lung (AFL), 10 with scleroderma with pulmonary manifestations (SCL), 9 with Wegener's granulomatosis (WG), and 7 with pulmonary Langerhans' cell histiocytosis (PLH). As a control, sera from 10 healthy volunteers were used. Neovascularization was measured by a leukocyte-induced angiogenesis assay according to Sidky and Auerbach. TNFalpha and INFgamma production was estimated by a one-step culture immunoassay CytoTraptrade mark TNFalpha DIA (Biosource Europe S.A.) after 3 h of incubation with LPS (TNFalpha) and 24 h incubation with PHA (INFgamma). Sera from sarcoidosis patients, WG patients, and AFL patients significantly stimulated angiogenesis in comparison with sera from healthy donors (P<0.001). Sera from PLH and SCL patients presented anti-angiogenic properties in comparison with sera from healthy donors and from each examined group (P<0.001). Comparing with other groups, preincubation with sera from AFL and WG patients led to a significant increase in TNFalpha production by normal MNC. Highly significant correlation between serum angiogenic activity and TNFalpha production by MNC was observed in SCL, WG, and AFL (r=0.74, P<0.01). we conclude that TNFalpha may play an important role in neovascularization in ILD.

Humoral immune response against mycobacterial antigens in children with tuberculosis.

Different clinical outcomes of tuberculosis (TB) are related to the balance between cell-mediated and humoral immunity and may depend on environmental and individual factors including age and sex. The purpose of this study was to analyze the humoral immune response to recombinant and native mycobacterial antigens in relation to clinical presentations of pulmonary TB in children. We examined 224 serum samples including 81 primary and 31 postprimary TB cases, 30 cases of latent TB infection, and 82 nontuberculosis controls. Commercially available ELISA assays detecting IgG, IgA, and IgM against antigens: 38 kDa, 16 kDa, 38 kDa, lipoarabinomannan (LAM), and A-60 were used. The results indicate that IgG production was very low in primary compared with postprimary TB (P<0.0001). IgM levels did not differ between the examined groups. Antibody levels strongly depended on the child's age. In infants aged below 1 there was no difference in the antibody level between the TB and control cases. Most positive cases were observed in children aged above 10. The influence of BCG vaccination on the antibody level was not seen. In all subgroups, person-to-person heterogeneity of antigen recognition was observed. We conclude that humoral immune response is associated with the phase of TB and is stronger in more advanced TB forms. IgG and IgA production against mycobacterial antigens is very low in young children.

Humoral immune response against mycobacterial antigens in bronchoalveolar fluid from tuberculosis patients.

Resistance to tuberculosis (TB) is cell-mediated but a humoral response is common and may be correlated with the lack of effective local cellular defense mechanisms. The goal of the study was to evaluate IgG, IgA, and IgM-mediated humoral immune response against 38-kDa+16-kDa and 38-kDa+lipoarabinomannan (LAM) mycobacterial antigens in bronchoalveolar fluid (BALF) from patients with pulmonary TB. Non-tuberculosis (NTB) patients were used as control. 179 BALF samples (56 TB and 123 NTB) were examined. Commercially available ELISA-based assays against proteins 38-kDa and 16-kDa or 38-kDa plus LAM were used. Three different dilutions of BALF: 1:1; 1:10, and 1:50 (100) were tested. Only the results obtained with the 1:10 dilution allowed distinguishing TB and NTB groups. The mean IgG level for 38-Da+LAM was significantly higher in the TB than that in the NTB group (P<0.0001). The mean IgA level for 38-kDa+LAM also was higher in the TB group (P<0.05). No difference was observed between TB and NTB groups in the titer of IgM antibodies. These findings indicate that TB is associated with the presence of detectable levels of antibodies in BALF. The antibody response is highly heterogeneous. This phenomenon results from the balance between pathogen and host immune system. The tests examined for detection of IgG in BALF can be used in combination with other diagnostic methods to increase diagnostic accuracy of pulmonary TB.

Diagnostic value of different serological tests for tuberculosis in Poland.

The aim of the study was to test the diagnostic accuracy of several serological assays for the diagnosis of tuberculosis (TB) in the Polish population. ELISA based assays detecting: 38 kDa+LAM - MycoM, MycoA and MycoG, 38 kDa - Pathozyme TB complex, 38 kDa+16 kDa - Pathozyme TB complex plus were used. The humoral immune response was analyzed in a group of 319 TB patients (289 adults and 30 children) and in a control group consisting of 66 sarcoidosis cases, 16 cases of mycobacterial infections other than tuberculosis, 35 lung cancer patients, and 70 healthy volunteers. Among the TB patients, there were 267 cases of pulmonary TB and 52 cases of extrapulmonary TB. Sensitivity varied between 32% (IgM) and 63% (IgA) and increased in culture positive tuberculosis and in chronic cases. Specificity was the highest for the tests based on recombinant antibodies (98%). Sensitivity of the IgG test in extrapulmonary TB was comparable with that in pulmonary TB. Overall, sensitivity of the examined tests was lower in children than in adults, but it varied depending on the age and phase of the disease. We conclude that the ELISA-based tests may be a useful tool for improving the diagnosis of TB, especially in adults and in those countries where the prevalence of culture positive and chronic cases is high.

Chocolate feeding of pregnant mice influences length of limbs of their progeny.

UNLABELLED: We previously reported the inhibitory effect of various methyloxantines and phenolic compounds on tumor-induced angiogenesis and the production of angiogenic growth factors. The aim of the present work was to evaluate the effect of chocolate (CH), food containing substantial amounts of methyloxantine theobromine and polyphenols (mainly catechins), given to mice during pregnancy and the lactation period, on weight of organs, length of limbs, and bone vascular endothelial growth factor (VEGF) concentration (tested by ELISA), in 4-week old offspring. The study was performed on 2-month old Balb/c mice fed during pregnancy and lactation 400 mg of CH daily. Content of polyphenols (catechines) and theobromine in the chocolate was estimated by high liquid perforance chromatography (HPLC). Concentration of VEGF was tested by ELISA. Feeding pregnant mice chocolate produced the following effects: decrease of relative length of limbs and thigh bones in 4-week old progeny and decrease in VEGF content of offspring femoral bones. CONCLUSION: attention should be paid to possible unwanted effects of catechine- and methyloxantine-rich food and beverages during pregnancy and lactation. 200 mg of chocolate per mouse corresponds to 100 g per person.

[Value of the immunochromatographic assay for detecting IgG antibodies against 38 kDa mycobacterial antigen in diagnosis of tuberculosis]. / Przydatnosc testu immunochromatograficznego wykrywajacego przeciwciala IgG przeciwko antygenowi pratka 38 kDa w rozpoznawaniu gruzlicy.

Despite of a fast development in the techniques of rapid identification of mycobacteria by molecular genetic techniques, serodiagnosis may be of special values as non-expensive, easy to perform method. Several serodiagnostic tests, principally those using immunoenzymatic (ELISA) methodology are available. The goal of our study was to evaluate one step coloured immunochromatographic assay detecting IgG antibodies against antigen 38 kDa (Rapid Test TB). Our material consisted of 278 serum samples--tuberculosis (n = 155), healthy (n = 36), sarcoidosis (n = 50), lung cancer (n = 25) mycobacterial infections other than tuberculosis (n = 12). Tuberculosis group consisted of new culture positive cases (n = 66), new culture negative cases (n = 23), chronic cases (n = 43) and extrapulmonary TB (n = 23). Specificity of 96% and sensitivity of 54% was obtained. In pulmonary TB sensitivity of 50% and in extrapulmonary TB of 74% was obtained. In chronic cases sensitivity of 70% and in new cases of 40% was received. Sensitivity of 44% in new culture positive cases and 30% in new culture negative cases was obtained. We conclude that immunochromatographic test may be a very useful tool improving tuberculosis diagnosis, especially in extrapulmonary tuberculosis. Strip test may be an interesting alternative as it is an extremely simple, rapid, and cheap technique.

Inhibitory effect of shark liver oil on cutaneous angiogenesis induced in Balb/c mice by syngeneic sarcoma L-1, human urinary bladder and human kidney tumour cells.

The effect of shark liver oil on cutaneous angiogenesis induced in mice by intradermal grafting of tumour cells was evaluated. It was shown that this substance (Ecomer) suppressed neovascular response in mice grafted with sarcoma L-1 syngeneic cells, human kidney cancer and human urinary bladder cancer cells. In addition, strong stimulatory effect of this drug on mice blood granulocyte number and their metabolic activity was observed.

Inhibitory effect of theobromine on induction of angiogenesis and VEGF mRNA expression in v-raf transfectants of human urothelial cells HCV-29.

Neovascularisation plays a crucial role in solid tumor growth and metastasis formation. Our previous studies showed that theophylline and theobromine suppressed cutaneous neovascular reaction induced in mice by human blood leukocytes, and lung as well as ovarian cancer cells. Here, we investigated the in vivo effect of theobromine on angiogenic activity of human urothelial cell line HCV-29, v-raf transfected (mouse cutaneous assay), and the in vitro effect of this drug on VEGF, tPA, uPA and PAI mRNA expression in these cells (RT-PCR method). Theobromine suppressed angiogenesis induced in mice by HCV-29-v-raf cells, inhibited VEGF mRNA expression, and had no effect on transcription of uPA and tPA in these cells. HCV-29-v-raf transfectants do not display transcripts of PAI, in the presence or the absence of theobromine.

[Diagnostic value of IgG antibody levels against 38 kDa mycobacterial antigen]. / Wartosc diagnostyczna poziomu przeciwcial IgG przeciwko antygenowi 38 kDa pratka gruzlicy.

Tuberculosis diagnosis bases on clinical and radiological symptoms and identification of mycobacteria. Accuracy of both methods is limited. Therefore reliable serological test would have considerable advantage. The present study was aimed at evaluating IgG-mediated immune response against specific mycobacterial antigens 38 kDa in group of 200 patients and control subjects. Our material consisted of 104 tuberculosis patients, 25 with sarcoidosis, 24 with lung cancer, 13 with bacterial or fungal pulmonary infection, 8 with mycobacterial infections other than tuberculosis and 26 healthy persons. We used commercially available ELISA based kits (Pathozyme TB-complex). Specificity of 100% and sensitivity of 49% was achieved. Sensitivity increased to 59% in chronic cases and to 52% in culture positive cases. Sensitivity decreased to only 14% in group of new culture negative cases. Measurement of IgG serum level against 38 kDa can be helpful in tuberculosis diagnosis. As the test lacks falsely positive results it indicates its high positive predictive value.