Angiogenic activity of sera from pulmonary tuberculosis patients in relation to IL-12p40 and TNFα serum levels.

The role of angiogenesis in the pathogenesis of tuberculosis (TB) is not clear. The aim of this study was to examine the effect of sera from TB patients on angiogenesis induced by different subsets of normal human mononuclear cells (MNC) in relation to IL-12p40 and TNFα serum levels. Serum samples from 36 pulmonary TB patients and from 22 healthy volunteers were evaluated. To assess angiogenic reaction the leukocytes-induced angiogenesis test according to Sidky and Auerbach was performed. IL-12p40 and TNFα serum levels were evaluated by ELISA. Sera from TB patients significantly stimulated angiogenic activity of MNC compared to sera from healthy donors and PBS (p < 0.001). The number of microvessels formed after injection of lymphocytes preincubated with sera from TB patients was significantly lower compared to the number of microvessels created after injection of MNC preincubated with the same sera (p < 0.016). However, the number of microvessels created after the injection of lymphocytes preincubated with sera from healthy donors or with PBS alone was significantly higher (p < 0.017). The mean levels of IL-12p40 and TNFα were significantly elevated in sera from TB patients compared to healthy donors. We observed a correlation between angiogenic activity of sera from TB patients and IL-12p40 and TNFα serum levels (p < 0.01). Sera from TB patients constitute a source of mediators that participate in angiogenesis and prime monocytes for production of proangiogenic factors. The main proangiogenic effect of TB patients' sera is mediated by macrophages/monocytes. TNFα and IL-12p40 may indirectly stimulate angiogenesis in TB.

[Diagnostic value of induced sputum in interstitial lung disease]. / Znaczenie diagnostyczne indukowanej plwociny w chorobach sródmiazszowych pluc.

BACKGROUND: Induced sputum (IS) has been recently used to investigate pulmonary inflammation in patients with interstitial lung disease (ILD), but still little attention has been paid to its efficacy in diagnosing sarcoidosis and other ILD. The aim of this study was to evaluate the diagnostic value of IS differential cell count and CD4+/CD8+ ratio in sarcoidosis (SA) and nonsarcoidosis ILD (NSA ILD). MATERIAL AND METHODS: We studied prospectively newly diagnosed 59 patients: 36 SA and 23 NSA ILD [16 hypersensitivity pneumonitis (HP) and 7 idiopathic pulmonary fibrosis (IPF)]. IS was performed by inhaling a 5% NaCl solution for 4 periods of 5 minutes. Giemsa stained cytopreps were differentially counted and T-lymphocyte subsets were analyzed by flow cytometry method. The k-nearest neighbour rule (k-NN) or predictive value of CD4+/CD8+ ratio were used to discriminate between SA and NSA ILD. The variables of IS used in k-NN rule were: cells viability, total cell count, percentages of alveolar macrophages, lymphocytes, neutrophils, eosinophils, CD4+ and CD8+ subsets, and CD4+/CD8+ ratio. RESULTS: 33 patients were able to produce an adequate sputum sample (SA-15, HP-11, IPF-7). A CD4+/CD8+ ratio>2.6 had a sensitivity of 100% and a specificity of 72% with 84% of correctly classified cases in distinguishing SA from NSA ILD. However, using k-NN rule the probability of correct classification was 79% (classification error rate-21%). CONCLUSION: To distinguish SA from NSA ILD cut off CD4+/CD8+ ratio>2.6 alone was superior to k-NN rule using all the parameters of IS.

Angiogenic activity of sera from extrinsic allergic alveolitis patients in relation to clinical, radiological, and functional pulmonary changes.

Extrinsic allergic alveolitis (EAA) caused by inhaled organic environmental allergens can progress to a fibrotic end-stage lung disease. Neovascularization plays an important role in pathogenesis of pulmonary fibrosis. The aim of this study was to assess the effect of sera from EAA patients on the angiogenic capability of normal peripheral human mononuclear cells (MNC) in relation to the clinical, radiological, and functional changes. The study population consisted of 30 EAA patients and 16 healthy volunteers. Routine pulmonary function tests were undertaken using ERS standards. As an angiogenic test, leukocyte-induced angiogenesis assay according to Sidky and Auerbach was used. Compared with sera from healthy volunteers, sera from our EAA patients significantly stimulated angiogenesis (P < 0.001). However, sera from healthy donors also stimulated angiogenesis compared to PBS (P < 0.001). No correlation was found between serum angiogenic activity and clinical symptoms manifested by evaluated patients. A decrease in DLco and in lung compliance in EAA patients was observed but no significant correlation between pulmonary functional tests and serum angiogenic activity measured by the number of microvessels or an angiogenesis index was found. However, the proangiogenic effect of sera from EAA patients differed depending on the stage of the disease and was stronger in patients with fibrotic changes. The present study suggests that angiogenesis plays a role in the pathogenesis of EAA. It could be possible that the increase in the angiogenic activity of sera from EAA patients depends on the phase of the disease.

[Comparison of Th1 and Th2 response in the blood of tuberculous patients and healthy contacts]. / Odpowiedz Th1-Th2 u chorych na gruzlice oraz osób zdrowych pozostajacych z nimi w stycznosci.

INTRODUCTION: Th1 response is known to play a dominant role in the resistance to tuberculosis. Nevertheless, IFN gamma levels are frequently increased in tuberculous patients, especially at the site of the disease. It is also possible that the shift toward Th2 response is responsible for the loss of resistance. The aim of this study was to compare the Th1 function of peripheral blood cells and the levels of antimycobacterial antibodies in the serum of culture positive tuberculosis patients and healthy tuberculosis (Tb) contacts. The correlation between the levels of antimycobacterial antibodies and Th1 function of blood cells was also evaluated. MATERIAL AND METHODS: The material consisted of 51 tuberculous patients and 20 healthy persons, close contacts of tuberculosis patients. The ability of peripheral blood cells to secrete IFN gamma and IL-2 was estimated in whole blood cultures with PHA, PWM and tuberculin. The levels of IFN gamma and IL-2 in the supernatants of cultures was estimated via a commercial ELISA test. The levels of antimycobacterial antibodies was measured with commercial immunoenzymatic kits detecting IgG antibodies against 38 kDa + 16 kDa and IgG, IgA and IgM antibodies to 38 kDa + lipoarabinomannan (LAM). RESULTS: No difference was found in the secretion of IFN gamma and IL-2 after stimulation with PHA and PWM between the patients and contacts. The secretion of IFN gamma after stimulation with tuberculin was even greater in tuberculous patients than in contacts. The levels of IgG and IgA (38 kDa + LAM) were higher in tuberculous patients than in contacts. There was a negative correlation between the level of IgG anti 38 kDa + LAM and the ability of peripheral blood cells to secrete IFN gamma after non-specific stimulation in patients with tuberculosis. CONCLUSIONS: Our study confirms the hypothesis that it is not the diminished production of Th1 cytokines, but rather the parallel overproduction of Th2 cytokines, which are essential in the development of tuberculosis.

[Evaluation of humoral immune response against mycobacterial antigens in bronchoalveolar lavage fluid from patients with pulmonary tuberculosis confirmed by genetic and culture methods]. / Ocena odpowiedzi humoralnej przeciwko antygenom pratkaw popluczynach oskrzelowo-pecherzykowych u chorych na gruzlicepluc potwierdzona bakteriologicznie i metodami genetycznymi.

INTRODUCTION: The resistance to TB is cells-mediated but humoral response is common and may be correlated with the lack of effective local cellular defence mechanisms. The goal of the study was to evaluate IgG, IgA and IgM mediated humoral immune response against 38kDa plus 16kDa and 38kDa plus lipoarabinomannan (LAM) mycobacterial antigens in BALF from patients with culture confirmed and PCR positive pulmonary tuberculosis (TB) compared to non-tuberculous controls (NTB). MATERIAL AND METHODS: 79 BALF samples (46 TB and 30 NTB) were examined. In 25 BALF samples from TB patients nucleic acids from M. tuberculosis were detected by PCR method. Commercially available ELISA - based assays against proteins 38kDa and 16kDa or 38kDa plus LAM were used. Three different dilutions of BALF: 1 : 1 and 1 : 10 were tested. Mean IgG level against 38 + LAM was significantly higher in TB group compared to control (p < 0,0001). No difference was observed between TB and NTB group in titer of IgM antibodies. RESULTS: Sensitivity of the tests based on IgG anti38kDa + 16kDa was 49%, IgG anti38kDa + LAM - 33%, IgA anti38kDa + LAM - 100%, IgM anti38kDa + LAM - 35%. Specificity of examined assays: IgA anti38kDa + LAM - 13%, IgM anti38kDa + LAM - 75%, IgG anti38kDa + 16kDa - 87%, IgG anti38kDa + LAM - 93%. The findings of the study indicate that TB is associated with the presence of detectable levels of antibodies in the BALF. CONCLUSIONS: Examined tests detecting IgG in BALF can be used in combination with other diagnostic methods to increase diagnostic accuracy of pulmonary TB.

[Humoral immune response against mycobacterial antigens in patients with tuberculosis and mycobacterial infections other than tuberculosis]. / Odpowiedz humoralna na antygeny pratka u chorych na gruzlice i mikobakteriozy.

The aim of the study was to compare humoral immune response against various mycobacterial antigens in TB and MOTT vs healthy control group. 350 serum samples from TB patients, 20 samples from MOTT patients and 58 samples from healthy donors were examined. ELISA detecting IgG, IgA and IgM against antigens: 38 kDa and 16 kDa, 38kDa and lipoarabinomannan, and A-60 were used. Mean IgG level was higher in TB compared to healthy controls (p<0,001). Mean IgG level against 38kDa and 38 + 16 kDa mycobacterial antigens was higher in TB than in MOTT group. Mean level of the IgG, IgA and IgM antibodies against LAM was higher in MOTT compared to TB patients. In all subgroups person-to-person heterogeneity of antigen recognition was observed. Humoral immune response to recombinant mycobacterial antigens significantly differs in TB and MOTT patients.

Angiogenesis and cardioprotection after TNFalpha-inducer-Tolpa Peat Preparation treatment in rat's hearts after experimental myocardial infarction in vivo.

The aim of the presented work was to evaluate whether short subcutaneous (s.c.) administration of TNFalpha-inducer-Tolpa Peat Preparation (TPP or TPP batch 0210) 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.

[Antimycobacterial antibody level in pleural, pericardial and cerebrospinal fluid of patients with tuberculosis]. / Ocena poziomu przeciwcial przeciwpratkowych w plynie oplucnowym, osierdziowym i mózgowo-rdzeniowym u chorych na gruzlice.

The goal of the study was to evaluate IgG, IgA and IgM mediated humoral immune response against 38kDa and 16 kDa or 38kDa and LAM mycobacterial antigens in pleural, pericardial or cerebrospinal fluid from patients with tuberculosis (TB) and to compare to non-tuberculous controls (NTB). 30 cerebrospinal fluids (CSF) (16 TB pts and 14 NTB pts), 17 pericardial fluids (6 TB and 11 NTB) and 20 pleural fluids (7 TB and 13 NTB) were examined. Commercially available ELISA-based assays (Pathozyme Tb complex plus, Myco G, A and M--Omega Diagnostic) were used. Tests were performed and cut off established according to manufacturer instruction. Mean IgG level against 38 + 16kDa was significantly higher in neurotuberculosis group compared to control (p<0.05). Sensitivity of the test in detecting neurotuberculosis was of 42% and specificity of 96%. Mean IgG, IgA and IgM against 38kDa + LAM level was higher in TB group compared to NTB in CSF. No difference was observed between TB and NTB group in pleural effusion. Antimycobacterial antibody levels were non-significantly increased in pericardial fluid in TB. The findings of the study indicate that TB is associated with the presence of detectable levels of antibodies in the CSF and pericardial effusion. Anti 38kDa + 16kDa IgG test can be used in combination with other diagnostic methods to increase diagnostic accuracy of neurotuberculosis.

[Proteinases inhibitors in sera of diabetic patients with non-proliferative retinopathy]. / Inhibitory proteaz w surowicach pacjentów z cukrzycowa retinopatia prosta.

Diabetes is associated with a hypercoagulable state. Eighty percent of patients with diabetes mellitus die due to various thrombotic vascular complications. Disorder of coagulation and fibrynolysis is associated with diabetic retinopathy and nephropathy. Angiogenesis requires degradation of vascular basement membrane prior to migration and proliferation of endothelial cells. Various serine proteases play important role in this process. The homeostatic system is also the source of endogenous inhibitors of angiogenesis. Human serum contains various factors able to induce or suppress formation of new blood vessels. The aim of the present study was to evaluate the activity of some angiogenesis inhibitors, anti-proteases, anti-thrombin III, a1 anti-trypsin and a2 anti-plasmin in sera of patients with diabetes mellitus type 1 and 2 and non-proliferative retinopathy, and to correlate this activity to total angiogenic potential of these sera, measured by mice cutaneous test. Sera of 22 persons with DM1, aged 33-70 years, 35 persons with DM2, aged 37-79 years, and 51 healthy people, aged 22-80 years (as control group) were studied. Direct serum-induced cutaneous angiogenesis test in mice (SIA) was applied. Berichrom (ade Behring) tests and immunoturbidimetric method were used for evaluation of anti-proteases activity. Angiogenic activity of DM1 patients sera was statistically lower than this parameter in DM2 patients and in control group. Levels of anti-proteases were similar in DM1, DM2 and control group, with one exception: anti-thrombin level was lower in DM2 patients' sera than this in the control group. Analysis of correlation revealed important difference in behaviour of DM1 sera, as compared to other groups. Significant negative correlation was observed between angiogenic activity and anti-thrombin, as well as anti-trypsin level of DM1 patients' sera. On the other hand, correlation analysis performed for the sera of control group revealed significant positive correlation between their angiogenic activity and anti-thrombin level. No other correlations were found.

[Usefulness of antibodies against A60 mycobacterial antigen in tuberculosis diagnosis]. / Przydatnosc oznaczania róznej klasy przeciwcial przeciwpratkowych anty-A60 w rozpoznawaniu gruzlicy.

Diagnosis of tuberculosis is based on clinico-radiological symptoms and on identification of mycobacteria. Accuracy of both methods is limited. Therefore reliable serologic test would have considerable advantage. The goal of our study was to evaluate serologic response against A60 mycobacterial antigen in group of 270 patients (pts) (137 pts with active tuberculosis (TB), 15 pts with mycobacterial infections other than TB (MOTT), 58 pts with sarcoidosis, 26 pts with lung cancer and 34 healthy controls. We used commercially available ELISA based kits against antigen A60 (ANDA-TB). Specificity of 87% for IgG, of 94% for IgA and for IgM, of 85% for IgG + IgA, of 89% for IgA + IgM, of 80% for IgG + IgM, and of 79% for IgG + IgA + IgM was achieved. Sensitivity of 40% for IgG, of 30% for IgA, of 18% for IgM, of 56% for IgG + IgA, of 35% for IgM + IgA, of 50% for IgG + IgM, and of 59% for IgG + IgA + IgM was obtained. The results demonstrated that humoral response differs depends on the immunoglobulin class. Measurements of IgG or IgG and IgA as most sensitive against A60 are potentially more useful serological tests in clinical practice. It is not recommended to use tests based on antigen A60 for differentiation tuberculosis and MOTT.