Calreticulin is a B cell molecular target in some gastrointestinal malignancies.

Calreticulin, upon translocation to the cell surface, plays a critical role in the recognition of tumour cells and in experimentally induced cellular anti-tumour immunity. However, less is known about anti-calreticulin antibodies and their role in malignancies. Using enzyme-linked immunosorbent assay (ELISA), we found immunoglobulin (Ig)A and/or IgG anti-calreticulin antibodies in sera of approximately 63% of patients with hepatocellular carcinoma (HCC), 57% of patients with colorectal adenocarcinoma (CRA) and 47% of patients with pancreatic adenocarcinoma (PACA), while healthy controls, patients with viral hepatitis C and with chronic pancreatitis reached only 2%, 20% and 31% seropositivity, respectively. We found significantly elevated mean levels of IgA anti-calreticulin antibodies (P < 0.001) in patients with HCC (78.7 +/- 52.3 AU, mean +/- standard deviation), PACA (66.5 +/- 30.9 AU) and CRA (61.8 +/- 25.8 AU) when compared to healthy controls (41.4 +/- 19.2 AU). Significantly elevated mean levels of IgG anti-calreticulin antibodies (P < 0.001) were detected in patients with HCC (121.9 +/- 94.2 AU), gall bladder adenocarcinoma (118.4 +/- 80.0 AU) and PACA (88.7 +/- 55.6 AU) when compared to healthy controls (56.7 +/- 22.9 AU). Pepscan analysis revealed a large number of antigenic epitopes of calreticulin recognized by both IgA and IgG antibodies of patients with HCC and PACA, indicating robust systemic immune response. Moreover, significantly elevated levels of antibodies against peptide KGEWKPRQIDNP (P < 0.001) in these patients, tested by ELISA, confirmed the distinct character of antibody reactivity against calreticulin. The high occurrence and specificity of serum anti-calreticulin autoantibodies in the majority of patients with some gastrointestinal malignancies provide the evidence for their possible clinical relevance.

[Calprotectin levels in patients with idiopathic inflammatory bowel disease comparison of two commercial tests]. / Stanovení koncentrace kalprotektinu ve stolici u nemocných s idiopatickými strevními zánety-srovnání dvou komercních testu.

BACKGROUND: Fecal calprotectin test is a simple, non-invasive, rapid and inexpensive diagnostic tool allowing differentiation between GIT functional disorders and inflammatory conditions and relapse prediction in non-specific inflammatory bowel disease. In the last year, commercially available ELISA diagnostic kits, using either monoclonal or polyclonal antibodies against a heterodimeric complex, calprotectin, for the detection of fecal calprotectin, started to be marketed. OBJECTIVE: To compare two ELISA kits for the detection of fecal calprotectin differing from each other in the used type of antibody (monoclonal versus polyclonal). MATERIAL AND METHODS: Two ELISA kits were assessed: Calprotectin ELISA (Bühlmann, Basel, Switzerland) using a monoclonal antibody against calprotectin and PhiCal Calprotectin ELISA (R-Biopharm, Darmstadt, Germany) using a polyclonal antibody against calprotectin. We analyzed fecal eluates from patients with Crohn's disease (CD, n=36) and ulcerous colitis (UC, n=29) and from healthy controls (n=98). Data were analyzed using software Statistica CZ 8.0 (Statsoft, Tulsa, U.S.A.) and measurement variability parameters (linearity, repeatability, stability) and test sensitivity and specificity were established and the methods were compared. RESULTS: The two kits showed adequate accuracy (intra- and inter-assay variation < 10%). The dilution linearity test indicated superiority of the Calprotectin ELISA Bühlmann kit, in particular for high calprotectin levels. The results of the two methods showed good correlation: Pearson's correlation coefficient r = 0.83, limit difference according to Bland-Altman plot ranged from 17% to 30%. Diagnostic sensivity rates were 79% for the Calprotectin ELISA Bühlmann kit and 74% for the Calprotectin ELISA R-Biopharm kit, the test specificity rates were 87% and 84%, respectively. CONCLUSIONS: Both of the tested kits have comparably good measurement parameters, the Bühlmann kit using monoclonal antibody against calprotectin showed higher sensitivity and specificity. In view of their availability, sensitivity and performance, the fecal calprotectin ELISA kits are helpful diagnostic tools for clinical practice.

IgE against food and respiratory allergens in healthy and allergic mothers and their children.

IgE against mixtures of common food or respiratory allergens were determined by ELISA in healthy (n = 38) and allergic (n = 62) mothers and their children. Significantly higher level of IgE against respiratory allergens was found in sera of allergic mothers and in cord blood of their children. No correlation between antibody level in maternal and newborn's sera was found; this argues against the transfer of IgE from mother to fetus and points rather to offspring's intrauterine sensitization. Specific IgE level in cord blood was higher in children who developed later allergy than in children who did not. Specific IgE level in colostrum was low both in healthy and allergic mothers; there was no correlation between high concentration of IgE against respiratory allergens in sera of allergic mothers and their colostrum, which does not support the idea of IgE transport from blood to mammary gland. Only slightly increased colostral IgE was detected in allergic mothers whose children manifested allergy later. Allergy of the mother and high level of anti-allergen IgE in her serum and in cord blood are the main predictive factors of future occurrence of allergy in the offspring. A combination of several predictive factors could have higher prognostic value.

Immunological profiles of patients with chronic myeloid leukaemia. I. State before the start of treatment.

In view of the increasing interest in the immunotherapy of CML it seems highly desirable to broaden the present knowledge on the immune reactivity of CML patients. A group of 24 patients and 24 healthy controls were studied for the total of 15 immunological parameters, including the prevalence of antibodies against human herpesviruses and papillomaviruses. To clearly discriminate between changes associated with the disease and those induced by the therapy, all patients were enrolled prior to the start of any anti-leukaemic therapy. Statistically significant differences between patients and controls were found in the levels of IgA, C4 component of complement, CRP and IL-6, the production of Th1 cytokines in stimulated CD3 cells and the E. coli stimulatory index. The analysis of the interrelationship between the results obtained in the individual patients presented some unexpected findings, such as the lack of correlation between the CRP and IL-6 levels. It will be the purpose of a follow-up to determine whether and how the immune status of the patients prior to the treatment correlates with their response to therapy and how the individual immunological profiles change in the course of the disease. These observations will be utilized in the future immunotherapeutic studies to constitute the vaccine- and placebo-treated groups.

[Comments on antiphospholipid antibody tests]. / Nekolik poznámek k laboratornímu vysetrováiní antifosfolipidoýsch protilátek.

The Lancet was the first to report the use of anticardiolipin antibody test in a group of patients at risk of recurrent arterial and/or venous thrombosis and recurrent pregnancy losses, 23 years ago. The condition characterized by specific clinical and laboratory signs and initially called the anticardiolipin syndrome came to be known as the antiphospholipid syndrome (APS) when cross-reactivity of cardiolipin with other phospholipids was revealed. The study of APS still arouses controversy. Even after two decades of research, there is disagreement on the standardisation and interpretation of antiphospholipid antibody (APLA) test results. More international workshops have been organized on APLA tests than on any other autoantibody test. However, there is still wide interlaboratory variation in APLA detection. Therefore, comprehensive quality control procedures have to be integrated into the routine workload of laboratories performing APLA analysis. Participation in an external quality assessment (EQA) scheme is essential for any laboratory seeking to maintain and provide quality service.

[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.

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.

[Autoimmune enteropathy with onset in early infancy: clinico-morphologic and immunologic manifestations]. / Autoimunní enteropatie se zacátkem v casném kojeneckém veku: klinicko-morfologické a imunologické projevy.

This paper describes a severely affected male infant with serious protracted diarrhoea caused by a rare autoimmune enteropathy. The disease began at 6 weeks of age of the child and it was associated with small bowel villous atrophy and the presence of circulating antienterocyte antibodies. The child was treated with steroids and with parenteral and special enteral nutrition. The patient showed clinical improvement as documented by decreased stool output and possibility to terminate the parenteral nutrition. The small biopsy samples showed a return to normal. Antienterocyte antibodies were negative after the treatment. The patient has been followed up for at least 18 months and was in a clinical remission. We recommend that autoantibodies tests should be performed in all infants with unexplained protracted diarrhoea. The use of potent immunosuppressive drugs and the increasing experience with parenteral and enteral nutrition can improve the perspective of these previously fatal disorders.

[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 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.

[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.

Influence of plasma exchange on serum levels of cytokines and adhesion molecules in ANCA-positive renal vasculitis.

BACKGROUND: Increased serum levels of proinflammatory cytokines may contribute to the organ damage in active antineutrophil cytoplasmic antigen (ANCA)-positive renal vasculitis. Plasma exchange (PE) may influence the activity of vasculitis not only by removing pathogenic autoantibodies, but also by lowering the serum levels of circulating cytokines. METHODS: Serum levels of IL-1beta, IL-1ra, IL-6, IL-8, ICAM-1 and VCAM-1 were measured using ELISA in 10 patients with active ANCA-positive renal vasculitis (5 patients with Wegener's granulomatosis, WG, and 5 patients with microscopic polyangiitis, MPA) during the course of therapeutic PE. Cytokines and adhesion molecules were measured in samples of serum obtained at the beginning and at the end of the 1st, 3rd and 5th PE and in samples of filtrate obtained during the same PE. RESULTS: In comparison with controls, patients with ANCA had higher serum levels of IL-1ra, IL-8, ICAM-1 and VCAM-1 before the 1st PE. Serum levels of IL-6, IL-8, ICAM-1 and VCAM-1 were increased in patients with MPA, and the serum levels of all the cytokines and adhesion molecules measured in patients with WG were increased. At the end of the PE course there were decreases in the serum levels of IL-1ra and VCAM-1 in ANCA patients and IL-1ra and ICAM-1 in WG patients. Single PE in ANCA patients led only to a decrease in serum levels of ICAM-1 and VCAM-1. On the other hand, there was no change in serum levels of IL-1beta and IL-8, and the serum levels of IL-1ra and IL-6 even increased at the end of a single PE, in spite of high levels of all cytokines and adhesion molecules in the plasma filtrate. CONCLUSION: Serum levels of soluble adhesion molecules decrease after PE, but serum levels of proinflammatory cytokines are not reduced even by a PE course. Removal of these substances by PE is obviously counteracted by their increased production, possibly further stimulated by the contact of blood with the synthetic membrane. The insufficient influence of PE on the elimination of proinflammatory cytokines may partially explain its limited effect in some patients with ANCA-positive renal vasculitis.

[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.

[The effect of plasmapheresis on cytokine serum levels and adhesion molecules in ANCA-positive renal vasculitis]. / Vliv plazmaferézy na sérové koncentrace cytokinu a adhezních molekul u ANCA-pozitivní renální vaskulitidy.

BACKGROUND: Increased serum levels of proinflammatory cytokines may contribute to the organ damage in active ANCA-positive renal vasculitis (ANCA-A). Plasma exchange (PE) may influence the activity of vasculitis not only by removal of pathogenic autoantibodies, but also by lowering of serum levels of circulating cytokines. METHODS AND RESULTS: Serum levels of IL-1, IL. 1ra, IL-6, IL-8, ICAM-1 and VCAM-1 were measured using ELISA in 10 pts with active ANCA-positive renal vasculitis (5 pts with Wegener's granulomatosis-WG, 5 pts with microscopic polyangiitis-MPA) during the course of therapeutic PE. Cytokines and adhesion molecules were measured in samples of serum obtained in the beginning and at the end of the 1st, 3rd and 5th PE and in the samples of filtrate obtained during the same PE. Pts with ANCA had before the 1st PE in comparison with controls higher serum levels of IL-1ra, IL-8, ICAM-1 and VCAM-1. There were increased serum levels of IL-6, IL-8, ICAM-1 and VCAM-1 in pts with MPA and increased serum levels of all measured cytokines and adhesion molecules in pts with WG. At the end of the course of PE there was the decrease of serum levels of IL-ira and VCAM-1 in pts with ANCA and IL-1ra and ICAM-1 in WG. Single PE led in pts with ANCA only to the decrease of serum levels of ICAM-1 and VCAM-1. On the other hand, there was no change of serum levels of IL-1 and IL-8 serum levels of IL-1ra and IL-6 even increased at the end of single PE, in spite of high levels of all cytokines and adhesion molecules in plasmafiltrate. CONCLUSIONS: Serum levels of soluble adhesion molecules decrease after PE, but serum levels of proinflammatory cytokines are not reduced even by the course of PE. Removal of these substances by PE is obviously counteracted by their increased production, possibly further stimulated by the contact of blood with synthetic membrane. Insufficient influence of PE on the elimination of proinflammatory cytokines may partially explain its limited effect in some patients with ANCA-positive renal vasculitis.

[Renal vasculitis and glomerulonephritis with anti-neutrophil cytoplasmic antibody positivity]. / Renální vaskulitidy a glomerulonefritidy s pozitivitou protilátek proti cytoplazme neutrofilních leukocytu.

BACKGROUND: Anti-neutrophil cytoplasmic antibodies (ANCA) define pathogenetically related group of renal vasculitides and glomerulonephritides mostly with serious prognosis. If unrecognized, these life-threatening diseases may cause loss of independent renal function and other dangerous extrarenal complications (e. g. pulmonary haemorrhage). We concentrated on the diagnosis, treatment and log-term follow-up of these patients. METHODS AND RESULTS: Renal biopsy was performed in 46 ANCA-positive patients. Age and sex distribution, type of ANCA, organ involvement, renal biopsy findings, renal function and effect of therapy were analyzed in these patients. Twenty three patients suffered from renal vasculitis, most commonly Wegener's granulomatosis (14 patients) and microscopic polyarthritis (7 patients). IgA nephropathy (7 patients) and idiopathic necrotizing/crescentic glomerulonephritis (8 patients) prevailed in patients with limited renal involvement. Renal morphology and function was most seriously impaired in patients with Wegener's granulomatosis. Immunosuppressive treatment was able to control the activity of the disease with the negativization of ANCA and improvement or stabilization of renal function in more than 90% of patients.CONCLUSIONS. ANCA-positive renal vasculitis and glomerulonephritis is relatively common. Clinical signs of extrarenal involvement are present in about 50% of patients with ANCA-positive glomerulonephritis. Rapidly introduced immunosuppressive treatment effectively controls renal and extrarenal manifestations of the disease.

Epstein-Barr virus serology in patients with autoimmune thyroiditis.

Elevated titers of antibodies against different antigens of Epstein-Barr virus (EBV) are found in some immunodeficient states, malignancies or in autoimmune disorders. We examined EBV serology in the group of 22 patients with autoimmune thyroiditis as compared with the group of 35 healthy volunteers. Titers of antibodies against viral capsid antigen (IgG-VCA) were more often found in the group of patients than in the control group (p = 0.000 35 for younger than 40 years and p = 0.00115 for older than 40 years) and the positivity of antibodies against early antigen (IgG-EA-D/DR) was also significantly more often found in the group of patients (p = 0.0031 and p = 0.0019 respectively) than in the control group.