[Immunoscore and Its Predictive Value for Colorectal Cancer]. / Immunoscore a jeho prediktivní hodnota u kolorektálního karcinomu.

Recent studies suggest that immune  classification (immune-score) in cancer patients has a prognostic value in some cases that seems to be superior to the AJCC/ UICC TNM  classification. The clinical outcome can vary significantly among patients with a particular diagnosis within the same TNM stage. Immunoscore methodology quantifies and detects different types of immune cells in tumor tissue, and also determines the density of their infiltration and localization at the tumor site. Currently within an international collaboration of 23 centers in 17 countries (including our department), immunoscore is being evaluated in more than 7,000 colorectal cancer patients in terms of the tumor microenvironment, focusing on the presence of immune cells both in the tumor tissue and the tumor invasive margin. Immunoscore results are assessed in correlation with: 1. patients response to the treatment, 2. rate of progression, disease prognosis and other immune parameters. It appears that the TNM classification and tumor invasiveness is statistically dependent on the immune response of the patient (there is an inverse correlation between the density of the infiltration of CD8⁺, CD3⁺ lymphocytes and the tumor stage). High densities of T-lymphocytes (CD8⁺, CD3⁺) both in the core and the invasive margin of the primary tumor are associated with longer term asymptomatic survival, overall survival, lower risk of relapse and reduced likelihood of metastases. The project of the international collaboration aims to introduce immunoscore in routine diagnostics.

Cellular and humoral immunodeficiency in breast cancer patients resistant to hormone therapy.

In view of the fact that insufficiency in immune response often correlates with poor prognosis, research in recent years has focused on the task of describing the precise status and function of the immune system and its possible effect on cancer patients. Although more than two thirds of treated patients respond to endocrine therapy, most patients with metastatic breast cancer develop a resistance to it. Estrogen modulates angiogenesis, partially through its effects on vascular endothelial growth factor (VEGF). It also appears that transforming growth factor-beta (TGF beta) could be another factor contributing to this resistance. TGF beta is a highly immunosuppressive factor that inhibits natural and specific immunity against tumors and stimulates the production of VEGF. The purpose of the study was to monitor immune responses in patients with hormone receptor-positive breast cancer who were resistant to hormone therapy. The examination of cellular components (CD4, CD8, HLA-DR, NK cells) and humoral immunity (IgG, IgG subclasses, IgA, IgM,). TGF beta and VEGF production were monitored with special attention, along with an analysis of the changes that occurred during the hormonal treatment. 68 patients included in the research project were implemented with routine cancer treatment with endocrine therapy. Basic parameters (the histological type and grade, the degree of expression of estrogen receptors (ER) and progesterone receptors (PR), human epidermal growth factor receptor 2 (HER2), and the proliferative marker) were established. Patients were evaluated by a cancer clinical immunologist to exclude immune disorders, allergic or autoimmune origin. TGF beta and VEGF were measured by ELISA and antitumor cellular immunity (CD4, CD8) was measured by flow cytometry. Patients who failed in the first line of hormone therapy treatment were considered as resistant to hormone therapy.Depression in cellular immunity was found especially in patients with resistance to endocrine therapy. In addition, immunoglobulin plasma levels were decreased (mainly IgG4 subtype). Most patients showed clinical symptoms of immunodeficiency (frequent infections of respiratory or urinary tract, herpetic infections). Significant increases in TGF beta and VEGF plasma were also detected.The correlation of these factors with resistance to hormonal therapy and the state of anticancer immunity could be helpful in the task of predicting resistance to hormonal therapy and could contribute to the selection of targeted immune therapy in cancer patients in the future.

Preoperative transforming growth factor-beta 1 (TGF-beta 1) plasma levels in operable breast cancer patients.

OBJECTIVES: The aim of this project was to search for new risk prognostic markers in the early stage of breast cancer. We tested preoperative plasma transforming growth factor - beta 1 (TGF- beta 1) levels in patients with operable breast cancer. Correlation with traditional prognostic markers and with positivity/negativity sentinel lymph node was evaluated. MATERIALS AND METHODS: Between 2003 and 2005, 36 patients with operable breast cancer (T1-2, N0-1, M0) with positive or negative sentinel lymph nodes were evaluated for their plasma TGF-beta 1. Twenty-seven healthy individuals (9 premenopausal and 18 postmenopausal) served as controls. Patients were evaluated for the traditional prognostic markers including tumor characteristics, positivity and negativity of sentinel lymph node, TNM, tumor grade, expression of tumor markers CA 15-3 and CEA, hormonal status (pre- or postmenopausal patients, estrogen and progesteron receptor expression), ERB and p53 expression. Predictive value of TGF-beta 1 level and correlation with either of the assessed parameters was tested by one way ANOVA analysis. RESULTS: Measurements of preoperative plasma TGF-beta 1 levels in patients with operable breast cancer were significantly higher compared with healthy individuals (median 15293 and 3983 pg/ml p < 0.0001). TGF-beta 1 level in plasma of patients with a positive sentinel lymph node was significantly higher than in patients with negative sentinel lymph nodes (high vs low, median 18,9 and 14,5 ng/ml, respectively, p = 0.05). CONCLUSION: The determination of TGF-beta 1 status might help to identify a high-risk population early in tumor progression, for which a more appropriate therapy should be established. In the node-negative population, the up-regulation of TGF-beta 1 might constitute an early event that promotes further progression of breast tumors.

[Transforming growth factor-beta 1 as a marker in patients with operable breast cancer]. / Transformacní rustový faktor--beta 1 jako marker u pacientek s operabilním karcinomem prsu.

UNLABELLED: OBJECTIVE AND DESIGN OF STUDY: Determination of TGF-beta 1 levels in plasma of patients with operable breast cancer. The correlation of TGF levels with the stage of disease and other prognostic markers. Prospective study. METHODS: There were 36 patients fulfilling the entrance criteria included in the study. The blood samples were taken to set the plasma levels of TGF-beta before the operation, with no adjuvant therapy. 27 age matched healthy volunteers served as controls. The study was approved by ethical board and patients signed informed consent regarding blood sampling and results presentation. Differences between groups were determined using the Mann-Whitney U-test. RESULTS: We proved that TGF-beta 1 levels are elevated in patients with operable breast cancer. Moreover, TGF-beta in plasma of patients with positive sentinel lymph node was significantly higher as compared to patients with negative sentinel node. Most important is the fact that patients involved in our study were in very early stages of disease. CONCLUSION: TGF-beta 1 is marker correlating with breast cancer disease risk factors, especially with positive sentinel lymph node. We conclude that TGF is one of the early markers which may help define the risk of disease progression already before the operation.

[Role of intestine in immunity mechanisms]. / Postavení streva v imunité.

The intestinal mucosa represents important primary interface with the external environment. The mucosal immunity posses the ability to differentiate between non-pathogenic microbial agents (non-inflammatory immune reaction) and pathogens (inflammatory immune response). The defence mechanisms of intestinal mucosa due to extensive nonadaptive and adaptive immune reactions facilitate both inductive and effector side of immune response. The oral tolerance is dependent on the mucous intestinal immunity and modulates the systemic immune reactivity.

[Antibodies against low density oxidized lipoproteins in type 2 diabetics]. / Protilátky proti oxidovaným lipoproteinum s nízkou hustotou u diabetiku 2. typu.

Oxidative modification of low density lipoproteins (LDL) is an important factor in the development of macrovascular atherosclerotic complications in patiens with type 2 diabetes mellitus. Recently autoantibodies against oxidized LDL (anti-oxLDL) have been suggested as a potential marker of LDL oxidation in vivo. The purpose of this study was to investigate the presence and levels of anti-oxLDL in patients with type 2 diabetes compared to healthy persons. We determined the serum concentrations of anti-oxLDL in 20 type 2 diabetic patiens with different degree and type of atherosclerotic vascular damage. Two healthy population groups: 20 young blood donors and 20 age and gender matched persons were used as controls. Anti-oxLDL positivity rates were distinctively higher in both control groups. Concentrations of anti-oxLDL were significantly lower in diabetic patients compared to both control groups. The incidence rates and levels of anti-oxLDL in both control groups were similar. Anti-oxLDL levels in the diabetes group did not correlate with the degree of macrovascular damage, serum total cholesterol, LDL cholesterol and triglyceride concentrations. We did not find any significant relationship between anti-oxLDL and other oxidative stress factors (superoxide dismutase, malondialdehyde, C and E vitamins). We suppose that anti-oxLDL may have an antiatherogenic protective role in healthy people but are not applicable to be an in vivo marker of LDL oxidation and macrovascular atherosclerotic vascular damage.

[Endothelial, phospholipid and oxidized LDL antibodies in patients with endogenous hypercortisolism]. / Protilátky proti endoteliím, fosfolipidum a oxidovaným LDL u pacientu s endogenním hyperkortizolismem.

Antiphospholipid (APLA), antiendothelial (AECA) and anti-oxidized LDL (anti-oxLDL) autoantibodies are found in vascular disorders. Pathogenetic contingency of atherosclerosis and these autoantibodies is still discussed, the mechanisms of their action in atherogenesis are not quite clear so far. Patients in various stages of endogenous hypercorticism as a model of accelerated atherosclerosis were investigated. We have sought possible correlations between autoantibodies and parameters of atherosclerosis with regard to the influence of endogenous hypercorticism on the inflammation. Low titres of autoantibodies in patients with active forms of disease result from the immunosuppressive effect of steroids. None of investigated group had high titres of APLA. No differences were found in AECA occurrence. No correlation of APLA, anti-oxLDL nor AECA with urinary free cortisol and plasma cortisol was found. There were no significant differences in autoantibody titres between patients with or without carotid stenosis. These results suggest, that autoantibodies may not always influence the development and progression of atherosclerotic lesions.

[Detection of the production of intracellular cytokines by T lymphocytes using flow cytometry--methodologic problems]. / Vysetrení produkce intracelulárních cytokinu T lymfocyty pomocí prutokové cytometrie--metodické problémy.

The method of assessment of intracellular proteins by means of flow cytometry makes it possible to evaluate the production of different cytokines by a clearly defined cell (sub-population type, state of cell activation). If the method should become a routine functional test, it must be standardized. This was the objective of our work when, based on data in the literature, we detected all controversial points and investigated them experimentally. Quite unequivocally we can recommend only sodium heparin as an anticoagulation agent when examining whole blood. The paper solves problems regarding the selection of mitogens where the marked effect of the use of mitogens on the result and necessity to compare results obtained under equal conditions was demonstrated. The authors tested also the possibility of preserving blood before processing and the selection of suitable combinations of surface signs and cytokines. When seeking the optimal time for cultivation it is necessary to make a compromise between the maximum possible production of cytokines (the kinetics of production of different cytokines is moreover different) and the accuracy of measurement because detection of the CD4 molecule after a prolonged period of stimulation deteriorates. As the optimum the authors recommend 4.5 hour cultivation with phorbol myristate acetate. The results proved a much greater capacity to retain newly formed cytokines in the cell if brefeldin A is used instead of monensine. The outcome of the work is a standard protocol for assessment of intracellular cytokines.

[Diagnostic value of determination of autoantibodies in gluten-sensitive enteropathy]. / Diagnostický prínos stanovení autoprotilátek u gluten-senzitivní enteropatie.

The study is focused on the immunopathological mechanisms of development of gluten-sensitive enteropathy (coeliac disease). It describes environmental factors and the role of autoantibodies and autoaggressive cells in the bowel inflammation. Attention is paid to the autoantibodies used in routine laboratory diagnosis of coeliac disease. The objective is a summary of rational diagnostic algorithms used in screening, diagnostics, treatment and dispensary care of patients with coeliac disease, especially with latent forms associated with other organ-specific immunopathological diseases. Exploration of anti-gliadin and anti-endomysial antibodies in diabetes mellitus type I were submitted. Furthermore, indications of these tests in the routine laboratory practice was analyzed.

T helper, cytotoxic T lymphocyte, NK cell and NK-T cell subpopulations in patients with chronic hepatitis C.

The phenotype of intrahepatic (IHL) and peripheral blood lymphocytes (PBL) was determined, and the production of cytokines by T lymphocytes analyzed in patients with chronic hepatitis C (CHC). Three-color fluorescence-activated cytometric analysis was done for 36 patients with untreated CHC. The percentage of peripheral blood memory T cells was higher in patients with CHC than in healthy controls (all data in %, significant at p < 0.001; 74.6 +/- 2.7 vs. 58.3 +/- 4.5), and a greater proportion of them were observed in the intrahepatic compartment (IHL-94.2 +/- 2.8 vs. PBL-74.6 +/- 2.7). There was a higher percentage of peripheral blood T helper 1 lymphocytes expressing IFN-gamma (IFN-gamma/IL-4) in these patients (4.6 +/- 0.7 vs. control-2.2 +/- 0.5). The expression of CXCR3 chemokine receptors on peripheral blood T helper cells was also high compared with the control (39.8 +/- 4.8 vs. 26.8 +/- 2.5) and a large percentage of T cells expressing CXCR3 or CCR5 chemokine receptors was observed in hepatitis C virus (HCV)-infected liver (CXCR3: IHL vs. PBL-74.9 +/- 5.7 vs. 39.8 +/- 4.8; CCR5: IHL vs. PBL-65.9 +/- 5.9 vs. 19.1 +/- 2.1). The intrahepatic compartment contains a greater proportion of activated cytotoxic T lymphocytes (CTL) and natural killer-T (NK-T) cells than peripheral blood (CTL: IHL vs. PBL-69.5 +/- 3.2 vs. 59.9 +/- 3.1; NK-T: IHL vs. PBL-10.6 +/- 2.5 vs. PBL: 3.99 +/- 0.5). The data suggest that in HCV-infected subjects, memory TH1 lymphocytes, activated CTL and NK-T cells compartmentalize in liver tissue and could play an important role in pathogenesis of chronic hepatitis.

The serologic screening for celiac disease in the general population (blood donors) and in some high-risk groups of adults (patients with autoimmune diseases, osteoporosis and infertility) in the Czech republic.

The prevalence of celiac disease (CD) was determined in healthy blood donors and in high-risk groups of adults (a total of 1835 adults--randomly selected 1312 healthy blood donors, 102 patients with primary osteoporosis, 58 patients with autoimmune diseases and 365 infertile women). It was calculated on the basis of a two-step serologic screening method--in the first step IgA and IgG antigliadin antibodies (AGA) and IgA anti-gamma-glutamyltransferase ('transglutaminase') antibodies (ATG) were estimated, in the second step sera positive for IgA AGA and/or IgA ATG were examined for antiendomysial IgA (AEA) antibodies. Immunoenzymic assay (ELISA) was used for determining of AGA and ATG antibodies; immunofluorescence method, performed on human umbilical cord tissue, was used for assaying of AEA antibodies. Total serum IgA level in only IgG AGA positive subjects was measured by routine turbidimetric method. 0.45% of healthy blood donors, 0.98% of osteoporotic patients, 2.7% of patients suffering from autoimmune disease and 1.13% of women with infertility considered as immunologically mediated were found to be positive in both steps of serologic screening (AGA and/or ATG and antiendomysium positive). The presumed high prevalence of seropositivity for CD in apparently healthy Czech adult population was confirmed. In the high-risk groups, the prevalence of seropositivity for CD was approximately 2-4 times higher than in healthy blood donors. The real prevalence of CD in the tested groups, however, can be estimated after performing small intestinal biopsy in the seropositive patients.

[Initial experience with detection of Saccharomyces cerevisiae antibodies in patients with primary nonspecific inflammatory bowel disease]. / Nase první zkusenosti s vysetrováním protilátek proti Saccharomyces cerevisiae (ASCA) u nemocných s primárními nespecifickými strevními zánety.

INTRODUCTION: Inflammatory bowel diseases (IBD), with Crohn's disease (CD) and ulcerative colitis (UC) as the two main disorders, is a heterogeneous group of diseases of unknown etiology. Actually we have no ideal disease marker, to identify people at risk of the disease, which can differentiate CD from UC, be highly specific for CD or UC and easily applicable in routine laboratory praxis. AIMS: Determine the clinical significance of serological testing p-ANCA and ASCA in patients with IBD. METHODS: P-ANCA in IgG isotype were detected by indirect fluorescence assay on human ethanol-fixed granulocytes, ASCA antibodies in IgG and IgA isotypes were determined by ELISA with mannan as a target antigen. RESULTS: P-ANCA and ASCA were studied in a group of 86 patients (38 CD, 26 UC, 3 non-inflammatory gastrointestinal disorder, 19 health controls). P-ANCA was associated with UC in 46%. ASCA was associated with CD in 76%. Specificity of ANCA for UC compared to healthy controls was 100%, specificity of ASCA for CD compared to healthy controls was 89.5%. CONCLUSION: Although the sensitivity of ASCA and p-ANCA is low, their specificity is high, especially when combining these two markers. We think that combined assay for ASCA and p-ANCA is more useful in IBD.

[The antiphospholipid syndrome and antiphospholipid antibodies]. / Antifosfolipidový syndrom a antifosfolipidové protilátky.

Antiphospholipid syndrome is a rate systemic autoimmune disease characterized by widespread arterial and venous thrombosis, recurrent abortion and thrombocytopenia. Laboratory tests reveal antibodies against phospholipids. These antibodies are detected by functional tests for the lupus anticoagulant, the anticardiolipin ELISA, the anti-beta 2-glycoprotein 1 ELISA and ELISA tests for antibodies against other cofactors and phospholipids. Pathogenetic mechanisms of thrombosis are poorly understood. Diagnostic assays for detection of antiphospholipid have not been yet adequately standardized.

Influence of cadmium and zinc sulphates on the function of human T lymphocytes in vitro.

Substances present in our environment influence the whole organism, including the immune system. Metals are one of the principal parts of these substances. It is generally supposed that they may have stimulating effect on immunity system in low concentrations in comparison with high concentrations in which they are toxic, with variations of each metal. There are differences between for example cadmium and zinc, and cadmium is toxic in low concentrations either. The effect of cadmium on the immune system has not been studied so deeply as the influence of zinc. In our work, we are interested in the study of immunomodulation caused by cadmium in comparison with the influence of zinc. We tested the effect of cadmium and zinc sulphates on human T lymphocytes in vitro. Molar concentrations of salts used in our work were from 10(-2) M to 10(-10) M. The influence of metals on lymphocytes in cell culture was studied by the expression of surface antigen CD69, blast transformation and IL-2 and IL-4 intracellular production after 2, 24 and 72 h cultivation. Cells were analyzed by flow cytometry using monoclonal antibodies. The results show more expressive differences in blast transformation. There are stimulating effects of cadmium in concentrations 10(-3) M, 10(-4) M and 10(-8) M, and zinc 10(-3) M. The most suppressive effect is in concentrations 10(-10) M of cadmium and 10(-8) M of zinc. The highest CD69 expression is in concentrations 10(-4) M to 10(-6) M of cadmium, and 10(-3) M of zinc. There are minimal differences in intracellular cytokine production in CD4+ lymphocytes effected by various metal concentrations used and between cadmium and zinc salts after 2 hours cultivation. There is the elevation of cytokines negative cells after the cultivation lasting 24 hours. Our investigation of metals' influence by different methods shows possibilities for further research.

[The chemiluminescence test in the laboratory evaluation of effects of immunostimulatory therapy]. / Vyuzití chemiluminiscencního testu pro laboratorní hodnocení úcinku imunostimulacní lécby.

BACKGROUND: Number of patients treated by general practitioners with various immunomodulatory drugs has recently increased. Effects of such medication on the immune system were not usually monitored. The aim of our work was to evaluate effect of selected immunomodulatory drugs on the phagocytic and metabolic activities of the phagocytes. METHODS AND RESULTS: 51 patients (18 males and 33 females) of the average age 36 years with repeating respiratory, mycotic and herpetic infections were investigated. Immunomodulatory treatment included: Decaris (Lavamizolum), Isoprinosine (Methisoprinolum), Imudon (Lysatum bacteriale mixtum), Biostim (Klebsiella pneumoniae), and Immodin (Leukocyti dialysati lyophylysatum). Before and after treatment all patients underwent basic immunological examination IgG, IgA, IgM, C3, C4 complement components, PEG, CD3, CD4, CD8 and CD19). Phagocytotic activity was estimated by means of FAGO MSHP test with HEMA particles and by chemiluminiscence test. Chemiluminiscence was measured using ML 3000 Microtiter Plate Luminometer (Dynex), 26 healthy individuals of the corresponding age were the controls. Results were statistically evaluated by Student's t-test. Significant increase of the cellular metabolic activity was found in Decaris and Immodin treated patients (P < 0.001). CONCLUSIONS: Chemiluminiscence test, which evaluates the metabolic activity of phagocytes, can be used for the accurate laboratory monitoring of the effects of some immunomodulatory drugs on the natural immunity of patients.

[Study of leukocyte phagocytic activity using flow cytometry and hydrophilic fluorochrome-labeled microspheric particles]. / Vysetrování fagocytární aktivity leukocytu na prutokovém cytometru mikrosférickými hydrofilními cásticemi znacenými fluorochromem.

The phagocytic activity was determined in human polymorphonuclear leucocytes using the flow cytometry technique with microspheric hydrophilic particles labelled with fluorescein isothiocyanate. Non-specific adherence of particles is discussed and normal values of phagocytic activity are presented. The advantages of this method include simplified sample preparation using whole blood, rapidity of the test, precise phagocytosis quantification and possibility of archiving all findings in a computer.

[Risk factors for ischemic heart disease in patients with chronic fatigue syndrome]. / Rizikové faktory ischemické choroby srdecní u pacientu s chronickým únavovým syndromem.

Risk factors of coronary artery disease (CAD) between a group of patients suffering of chronic fatigue syndrome (CFS) and a control group of healthy persons (whose exercise activity was not health-limited) were compared. Thirty three patients (27 women, 6 men, average age 39.9 +/- 11.7 years) and the same number of controls matched in age (39.8 +/- 10.3 years), gender and body weight. The Minnesota Questionnaire (by Taylor) and the Compendium of Physical Activities (by Ainsworth) were used to estimate total energetic expenditure in exercise activity as well as in job. The risk factors of CAD in the patients with CFS were not higher than in the control group. Aerobic physical fitness, basic anthropometric data, blood pressure, spectrum of blood lipoproteins, blood uric acid and smoking habits were not different between the compared groups. Patients suffering from CFS had lower total energetic expenditure in exercise activity. Nevertheless, this significant difference in sports activity was not large enough to cause any difference in risk factors of CAD between the CFS patients and the control group.

Cytokines and adhesion molecules in renal vasculitis and lupus nephritis.

BACKGROUND: Plasma levels of some pro-inflammatory cytokines and soluble adhesion molecules have been suggested to be useful parameters to assess the activity of antineutrophil cytoplasmic antibody (ANCA)-positive vasculitis and lupus nephritis. We hypothesized that the renal activity of these diseases is better reflected by the urinary excretion and fractional excretion of these molecules. METHODS: Plasma levels and urinary excretion of tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, IL-8, and the soluble cell adhesion molecules sICAM-1 and sVCAM-1 were measured by enzyme-linked immunosorbent assay (ELISA) in 14 patients with ANCA-positive renal vasculitis (eight active, ANCA-A; six in remission, ANCA-R), six patients with active lupus nephritis (LN), 15 patients with IgA nephropathy (IgAN) and nine healthy subjects. Fractional excretion of selected cytokines and adhesion molecules was also calculated. RESULTS: Patients with ANCA-A had increased urinary excretion and fractional excretion of TNF-alpha (9.27 +/- 3.19% vs 0.58 +/- 0.02%, P < 0.01), IL-6 (120.79 +/- 65.83% vs 1.89 +/- 0.34%, P < 0.01) and increased fractional excretion of IL-8 (23.34 +/- 6.38% vs 2.56 +/- 1.07%, P < 0.01) and sVCAM-1 (0.81 +/- 0.33% vs 0.03 +/- 0.02%, P < 0.01) compared with controls. Urinary excretion of TNF-alpha and IL-6 and fractional excretion of TNFalpha, IL-6 and IL-8 were higher in ANCA-A than in ANCA-R. Patients with LN had increased plasma TNF-alpha (20.52 +/- 2.01 pg/ml vs 12.33 +/- 0.23 pg/ml, P < 0.05) and sVCAM-1 (1537.88 +/- 276.36 ng/ml vs 692.26 +/- 44.42 ng/ml, P < 0.05) and increased urinary excretion of TNF-alpha (2.81 +/- 0.51 microg/mol creat vs 0.98 +/- 0.05 microg/mol creat, P < 0.01), IL-8 (35.78 +/- 14.03 microg/mol creat vs 12.46 +/- 5.19 microg/mol creat, P < 0.05) and sVCAM-1 (48.98 +/- 20.20 microg/mol creat vs 2.92 +/- 1.35 microg/mol creat, P < 0.01) compared with controls. Patients with IgAN had, in comparison with controls only increased plasma TNF-alpha (18.10 +/- 0.57 pg/ml vs 12.33 +/- 0.23 pg/ml, P < 0.05). CONCLUSIONS: Urinary excretion and fractional excretion, but not plasma levels, of selected pro-inflammatory cytokines (TNF-alpha, IL-6 and IL-8) were increased in patients with active ANCA-positive renal vasculitis, but not in ANCA positive vasculitis in remission. These parameters may be useful to monitor the activity of this disease.

[Chronic fatigue syndrome]. / Chronický únavový syndrome.

A great concern is recently given to the chronic fatigue syndrome in the Czech Republic. Unfortunately, published data allow us to state neither the etiologic agent nor the pathophysiology of the disease. Although many authors published various laboratory abnormalities, these changes are inconstant and do not allow to state a diagnosis of the chronic fatigue syndrome by a single laboratory test, and effective therapy is not known either. Psychotherapy, and in some cases antidepressants, are recommended by some authors to alleviate patient's symptoms. Neither immunological nor antiviral therapy showed positive results in controlled trials and are not generally used in most centers.

[CD5-positive B lymphocytes and antinuclear antibodies]. / CD5 pozitivní B lymfocyty a antinukleární protilátky.

BACKGROUND: The production of natural autoantibodies incl. antinuclear antibodies (ANA) is ascribed to lymphocytes which have a CD5 molecule on their surface. The role of CD5 positive B lymphocytes in the induction of autoimmunity is obscure so far. The authors focused their attention on the incidence of antinuclear antibodies (AA) in subjects with different diseases and sought a relationship with the ratio of CD5 positive B lymphocytes in the peripheral blood stream. METHODS AND RESULTS: CD5 positive lymphocytes were assessed on a flow cytometer using monoclonal anti CD5 and anti CD19 antibodies. Antinuclear antibodies are detected by indirect fluorescence on a substrate of human leucocytes and HEP-2 cells. In a group of 134 subjects the authors did not provide evidence of a direct relationship between the relative number of CD5 positive B lymphocytes in the peripheral blood stream and the presence of ANA (IgG, IgA, IgM), not even in 33 patients with autoimmune diseases. In 86 patients the authors revealed that antinuclear antibodies type IgM predominate in patients with repeated respiratory infections. In systemic diseases the isotype IgG predominates (p = 0.01). After immunosuppressive treatment with a favourable clinical effect the ANA isotype IgG disappears and isotype IgM is found more frequently. The incidence of the ANA isotype IgM is significantly higher in healthy subjects aged over 60 years than in younger subjects (12.5%/6.1%, p = 0.06), and more frequent in women (p = 0.05). CONCLUSIONS: The presence of antinuclear antibodies is not associated with the amount of CD5 positive B lymphocyte in the peripheral blood stream.