The role of angiogenesis in the pathogenesis of interstitial lung diseases (ILD) is unknown. Angiotensin-converting enzyme (ACE) is a marker of sarcoidosis activity and may modulate angiogenesis. The aim of this study was to examine the relationship between ACE activity in ILD patients' sera and their effect on microvessels formation in an in vivo model of leukocyte-induced angiogenesis. The study population consisted of 77 sarcoidosis patients, 22 idiopathic pulmonary fibrosis patients, 16 bird fanciers lung patients, eight silicosis patients and 14 healthy donors. Serum ACE activity was assayed by spectrophotometric method. As an angiogenic test, a leukocyte-induced angiogenesis assay in an animal model was used. Sera from interstitial lung disease patients significantly stimulated angiogenic activity of mononuclear cells compared with healthy donors (p < 0.001). The highest ACE serum activity was measured in sera from the silicosis patients, and lowest in sera from the sarcoidosis and IPF patients. A significantly lower serum ACE activity was detected in the bird fanciers lung patients. Serum angiogenic activity of ILD patients measured by angiogenesis index negatively correlated with ACE serum activity (r = ;-0.52; p < 0.01). This correlation was highest in the sarcoidosis group (r = -0.6; p < ). Sera from ILD patient constitute the source of factors modulating angiogenesis.

Chronic inflammation stimulates of neovascularization. The aim of this study was to evaluate the effect of sera from interstitial lung diseases (ILD) patients on angiogenic capabilities of different subsets of mononuclear cells. Serum samples were obtained from 22 patients with sarcoidosis, 20 with hypersensitivity pneumonitis, 20 with idiopathic pulmonary fibrosis, 9 with systemic sclerosis, 6 with pulmonary Langerhans cells histiocytosis, and from 20 healthy volunteers. Animal model of leukocyte induced angiogenesis assay was used as an angiogenic test. The pattern of angiogenic reaction was different in different diseases. Sera from systemic sclerosis and pulmonary Langerhans cells histiocytosis patients exerted inhibitory effects on angiogenesis, but sera from sarcoidosis, hypersensitivity pneumonitis, and idiopathic pulmonary fibrosis patients stimulated angiogenesis. Sera from sarcoidosis and pulmonary Langerhans cells histiocytosis primed monocytes for the production of angiogenic factors. The number of microvessels created after incubation of mononuclear cells depleted of monocytes with sera from systemic sclerosis patients significantly decreased. We conclude that the role of monocytes in the modulation of angiogenesis varies depending on the type of ILD. Sera from sarcoidosis stimulate and from pulmonary Langerhans cells histiocytosis patients inhibit neovascularization induced by monocyte mediators. Sera from systemic sclerosis inhibit angiogenesis induced by lymphocyte products.

OBJECTIVE:Clinical symptoms and radiological changes are useful in monitoring patients with interstitial lung diseases (ILD). Neovascularization participates in the pathogenesis of idiopathic pulmonary fibrosis and other ILD. The objective of the study was to examine the relationships between angiogenic activity of sera from ILD patients and clinical or radiological status.MATERIAL AND METHODS:Serum samples were obtained from 83 patients with sarcoidosis, 31 with idiopathic pulmonary fibrosis (IPF), 29 with hypersensitivity pneumonitis (HP), 16 with collagen diseases with pulmonary manifestation (CD), 13 with scleroderma (SCL), 14 with Wegener's granulomatosis (WG), 12 with pulmonary Langerhans cell histiocytosis (HIS), 12 with pneumoconiosis (PNC), 10 with drug-induced lung disease (DLD), 5 with cryptogenic organizing pneumonia (COP), and from 36 healthy volunteers. As an angiogenic test we used a cutaneous angiogenesis assay according to Sidky and Auerbach. Clinical status was evaluated using a special questionnaire. In all patients chest radiographs were performed.RESULTS:The angiogenic properties of sera from ILD differed depending on the clinical diagnosis. The strongest proangiogenic effect was induced by sera from patients with HP (mean number of new vessels 16.8), CD (16.6), sarcoidosis (16.3), IPF (16.2), and PNC (15.7). In the case of DLD (13.2), the effect was comparable to healthy controls (13.5). In contrast, sera from SCL (mean number of the vessels 10.5) and HIS patients (10.8) significantly inhibited angiogenesis compared with controls. The angiogenic activity of sera from patients with hilar or mediastinal lymph nodes involvement was higher than that of sera from patients with lung fibrosis. There were also differences in the serum angiogenic activity in relation to the severity of dyspnea.CONCLUSIONS:The data showed that sera from ILD patients constitute a source of mediators modulating angiogenesis, but the pattern of reaction is different in various diseases. Sera from HP, sarcoidosis, IPF, and CD patients demonstrated the strongest proangiogenic activity. However, sera from SCL and HIS inhibit angiogenesis. Angiogenic activity of examined sera was related to the clinical and radiological changes.

It is not clear why some individuals with unknown predisposition develop tuberculosis, while others remain healthy in spite of heavy exposure. Interferon gamma (IFNgamma) is considered to be the key cytokine responsible for resistance to M. tuberculosis infection, as confirmed by increased susceptibility to mycobacterial infections in rare inherited defects in IL-12-IFNgamma axis. The aim of this study was to assess the IFNgamma production by peripheral blood lymphocytes from immunocompetent tuberculosis (TB) patients. The study group included 51 TB patients. In all cases, TB was confirmed by culture. Twenty healthy TB contacts were considered as control group. Commercially available ELISA-based assays were used to measure IFNgamma in the supernatant of whole blood cell cultures after stimulation with PWM (Phytolacca Americana), PHA (phytohemagglutynin), and PPD (purified protein derivative). No difference in IFNgamma secretion between the patients and control group was found when blood cells were stimulated by PWM or PHA. PPD-induced IFNgamma formation was higher in TB patients than in controls. The secretion of IFNgamma after non-specific stimulation varied in different clinical and radiological presentation of tuberculosis and it was lower in most advanced and extensive forms of the disease. It is unclear whether the difference in formation and release of IFNgamma is a primary or secondary phenomenon in the course of the disease.

Health care workers (HCWs) are at risk for developing active tuberculosis (TB). The prevalence of latent tuberculosis infection (LTBI) in this group is unknown in Poland, due largely to the problems associated with the interpretation of the tuberculin skin test (TST) in BCG immunized population. The goal of the present study was to assess the prevalence of LTBI in both clinical and non-clinical 155 HCWs (120 females and 35 males) and to compare the groups at different levels of risk. All participants were interviewed using a questionnaire and underwent interferon-gamma whole blood assay (Quantiferon-Tb-Gold) (QTF) and TST. The questionnaire provided information on possible risk factors for LTBI, including demographic and socioeconomic details, the presence of BCG scars, and the degree of occupational exposure. We found that the prevalence of LTBI among HCWs was, on average, 27.1%. A higher risk of acquiring LTBI was associated with certain work locations (TB lab workers--prevalence 50%, TB ward clinicians--34%, nurses--30%). The prevalence in analytical lab technicians was 20%, in administration staff was 15%. The HCWs with positive QTF test results were older and worked longer than those who had negative results. There was a significant correlation between the level of IFN-gamma and both age (P<0.001) and length of employment (P<0.01). The correlation between the diameter of skin test induration and the magnitude of the INF-gamma response also was significant (P<0.001). We conclude that HCWs are at increased risk of infection, suggesting that appropriate preventive strategies should be undertaken. IFN-gamma test is a useful tool in detecting LTBI cases in a country where BCG vaccination is a national policy.

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.

Induced sputum is a useful non-invasive method for the assessment of airway and parenchymal lung diseases. This study aimed to compare induced sputum and bronchoalveolar lavage fluid (BALF) cellular composition and T-lymphocyte subpopulations in patients with interstitial lung disease. We evaluated 33 patients: 15 with sarcoidosis, 11 with hypersensitivity pneumonitis, and 7 with idiopathic pulmonary fibrosis. The percentage of macrophages was significantly lower in induced sputum than in BALF in sarcoidosis (P=0.005), and the percentage of neutrophils was higher in induced sputum than in BALF in sarcoidosis (P=0.001) and hypersensitivity pneumonitis (P=0.006). A significant correlation was found between the BALF and induced sputum CD4+, CD8+ subsets and the CD4+/CD8+ ratio in both the whole patient group (r(s)=0.80, r(s)=0.88, r(s)=0.88, P<0.001, respectively) and in the 3 subgroups. A strong correlation of the T-lymphocyte subsets in induced sputum and BALF in patients with interstitial lung disease shows that induced sputum may be a non-invasive surrogate for certain parameters in BALF in these patients.

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.

Different clinical outcomes of tuberculosis can be related to the balance between cell-mediated and humoral immunity. In this prospective study we examined the humoral immune responses to recombinant and native mycobacterial antigens in relation to clinical presentations of pulmonary TB. Two hundred and fifteen serum samples were examined including: non-cavitary (n=120), cavitary (n=65), caseous pneumonia (n=12), and disseminated TB (n=18). ELISA tests detecting IgG, IgA, and IgM against antigens: 38 kDa and 16 kDa, 38 kDa and lipoarabinomannan (LAM) were used. Univariate and multivariate logistic regression analyses were carried out to find the association between the antibody level and demographic or clinical characteristics. The relationships among specific antibody profiles and the phase of the disease in relation to demographic (age and sex) and clinico-radiological factors were investigated by measuring serum antibody levels (IgG, IgA, and IgM) to 38 kDa and 16 kDa recombinant M. tuberculosis antigens and to LAM - native mycobacterial antigen. The results show that the radiological extent of the disease is the strongest factor associated with IgG antibody production. Patients with more extensive pulmonary TB showed higher titers of IgG antibody to M. tuberculosis antigens (P<0.0001). The highest IgG and IgA level were observed in fibro-cavernous TB. The presence of cavity was associated only with IgG anti 38+16 kDa (P<0.001). IgA level was the highest in caseous pneumonia. IgM antibody production was not associated with any clinical and radiological factor, but only with the male gender. Age was independently and inversely associated with IgG anti 38 kDa+LAM level and IgM anti 38 kDa+LAM. We conclude that the humoral immune response to mycobacterial antigens is highly heterogeneous and varies with the stage of TB. IgG antibody level is higher in most advanced and extensive forms of the disease.

Sarcoidosis (SAR) is a systemic granulomatous inflammatory disease characterized by recruitment and activation of peripheral blood mononuclear cells to the sites of disease. Neovascularisation is a principal vascular response in chronic inflammation and hypoxia. The aim of the study was to evaluate the effect of sera from sarcoidosis patients on angiogenic capability of different subsets of normal peripheral human mononuclear cells (MNC) in relation to IL-6 and IL-8 serum levels, to radiological stages of disease and to the presence of extrapulmonary changes. Serum samples obtained from 42 sarcoidosis patients were examined. There were 12 patients in stage I, 16 patients in stage II, and 14 in stage III. In order to quantify angiogenesis, a leukocyte-induced angiogenesis assay was performed by a method of Sidky and Auerbach. MNC were depleted in monocytes by glass adherence and phagocytosis of iron particles techniques. IL-6 and IL-8 in sera from sarcoidosis patients were evaluated by an ELISA-based assay. Sera from sarcoidosis patients enhanced angiogenic capability of normal MNC significantly stronger than sera from healthy donors (P<0.001). Angiogenic activity of sera in sarcoidosis depended on the stage of disease and appeared most pronounced in stage II (P<0.05). Sera from patients with extrapulmonary changes exerted stronger effect on angiogenesis than sera from patients with thoracic changes only (P<0.001). IL-6 and IL-8 serum level correlated with each other, but no correlation was found between IL-6 and IL-8 serum level and angiogenic activity of the examined sera. Removal of monocytes from MNC eliminated the effect of sera from sarcoidosis patients on angiogenesis compared with the effect of these sera on intact MNC (P<0.001). Sera from sarcoidosis patients and from healthy people constitute a source of mediators participating in angiogenesis. Sera from sarcoidosis patients prime monocytes for production of proangiogenic factors.

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.

Different clinical outcomes of tuberculosis (TB) are related to the balance between cellmediated 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, 38kDa, 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.

The aim of the presented work was to evaluate whether short subcutaneous (s.c.) administration of TNF alpha-inducer-Tolpa Peat Preparation (TPP or TPP batch 0210)(R) modulates the process of ischemic remodeling and spontaneous angiogenesis after experimental myocardial infarction (MI) in rats in vivo. The results obtained using three complementary and correlative methods: histological studies, Proliferating Cell Nuclear Antigen (PCNA) reaction and Lymphocytes Induced Angiogenesis (LIA) test showed a clear pro-angiogenic and cardioprotective effect of TPP administration after experimental MI.TPP batch 0210 should be considered as an angiogenesis stimulating factor and consecutively as a cardioprotective preventing development of ischemic cardiomyopathy after MI in rats.It might possibly be used as an adjunct to conventional therapy of coronary artery disease, including late phase after myocardial infarction or ischemic cardiomyopathy. (c) 2005 Elsevier Inc. All rights reserved.

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.

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.

Background: Chronic obstructive pulmonary disease (COPD) is accompanied by both airway and systemic inflammation and by oxidative stress. This study aimed to characterise the relationship between oxidative stress and inflammatory components in induced sputum and blood.Material & methods: We studied blood and sputum samples from stable COPD patients (mean FEV1 60.5+/-7.5% predicted) at baseline (no treatment) and after 10 weeks treatment with either inhaled steroid, fluticasone propionate (FP) (1000 mug/d) or 10 weeks treatment with N-acetylcysteine (600mg/d) (NAC). We assessed the inflammatory markers (IL-8, ECP, sICAM-1, NE) in sputum and serum and we compared them with blood markers of oxidative stress (SOD, GPx, TEAC, albumin, vitamin E and A).Results: At baseline blood sICAM-1 correlated with IL-8 levels (P<0.01, r = 0.62) and negatively with GPx (P<0.01, r = -0.63) and with TEAC (P<0.05, r = -0.53). TEAC correlated positively with'GPx (P < 0.01, r = 0.70). dCorrelation between sICAM and IL-8 disappeared after NAC treatment. The correlation between sICAM and GPx disappeared after FP treatment. The correlation between TEAC and GPx was maintained after both NAC and FP.Conclusions: The relationship between markers of inflammation, adhesion and antioxidant capacity is significantly modulated by treatment with N-acetylcysteine or inhaled corticosteroids. (C) 2004 Elsevier Ltd. All rights reserved.