Impact of interleukin 13 (IL13) genetic polymorphism Arg130Gln on total serum immunoglobulin (IgE) levels and interferon (IFN)-γ gene expression.

This cross-sectional study was designed to investigate the extent of genetic susceptibility by targeting variants in interleukin (IL)-4/IL-13 signalling pathways leading to atopic disease in early childhood. We evaluated involvement of five single nucleotide polymorphisms IL4 C-590T, IL13 C-1055T, IL13 Arg130Gln, IL4RA Ile50Val and IL4RA Gln576Arg, in the control of serum total and antigen-specific immunoglobulin (Ig)E levels. Furthermore, we analysed their association with changes in gene expression of five cytokines having key roles in inflammatory and anti-inflammatory immune response [IL-4, IL-13, interferon (IFN)-γ, IL-8 and IL-10]. Total and antigen-specific IgE levels in serum and gene expression of selected cytokines in peripheral blood were measured in 386 children aged 1-8 years. TaqMan allelic discrimination, amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and restriction fragment length polymorphisms (RFLP) methods validated by sequencing were used for genotyping. All genotypes for children with total and antigen-specific IgE levels in the normal range were in Hardy-Weinberg equilibrium. Gene expression analyses were carried out using TaqMan gene expression assays. We found elevated total IgE levels in carriers of IL13 Arg130Gln variant allele [odds ratio (OR) = 1·84; 95% confidence interval (CI) = 1·16-2·93]. This effect was more apparent for boys (OR = 2·31; 95% CI = 1·25-4·28). However, no significant association was observed for the other four variants examined. We found up-regulation of IFN-γ in children with elevated serum total IgE levels carrying the Arg130 allele (P = 0·005). No differences were found for IL4, IL8 or IL10, while IL13 gene expression was under the detection limit. IL13 Arg130Gln genotypes can play a role in genetic susceptibility to allergy via regulation of serum total IgE levels and affecting IFN-γ gene expression.

The kinetics of cell surface receptor expression in children perinatally exposed to polychlorinated biphenyls.

Exposure to polychlorinated biphenyls (PCBs) during pre-natal and early life can alter normal immune system development. Blood specimens from newborns, 6-, and 16-month-old infants were collected in the Michalovce and Svidnik/Stropkov districts, areas with, respectively, high and low environmental PCB contamination, and lymphocyte receptor expression was evaluated by multi-color flow cytometry. The results indicate that the percentage of lymphoid dendritic cells (DC) and naïve/resting T-lymphocytes were significantly increased at 6-months in Michalovce as compared to the same cell types in cord blood samples (p < 0.001), whereas natural regulatory T-lymphocytes and suppressor inducer T-lymphocytes were reduced (p < 0.001). Overall, a positive linear correlation of terminally differentiated effector memory (TEM) T-lymphocyte population with age, but a negative linear correlation for myeloid DC from birth to 6-months in both regions were found. Michalovce samples indicated significantly higher expression of memory T-lymphocytes (birth, 6(th), and 16(th) month), TEM T-lymphocytes (birth and 6(th) month), and lymphoid DC (6(th) month) compared to the Svidnik/Stropkov regions. After adjustment for relevant covariates, such as maternal age, parity, season of birth, breastfeeding, birth weight, and gender, the myeloid DC, suppressor inducer T-lymphocytes, truly naïve helper/inducer T-lymphocytes, and TEM T-lymphocytes remained significantly different between districts in cord blood samples. The multivariate analysis models for 6- and 16-month samples showed district differences in all cellular determinants, except for lymphoid DC and macrophage-like cells. This study provides the first evidence that pre-natal and early post-natal exposure to PCBs affects the dynamics of cell surface receptor expression on lymphoid DC and DC-like cells, suggesting impaired immunologic development following pre-natal and early post-natal PCB exposure.

Dynamics of lymphocyte subsets in children living in an area polluted by polychlorinated biphenyls.

Immune system development, particularly in the pre-natal and early post-natal periods, has far-reaching health consequences during childhood, as well as throughout life. Exposure to poly-chlorinated biphenyls (PCBs) during pre-natal and early life has been previously associated with changes in the incidence of infectious and allergic diseases in children, and humoral immunity alterations. Lymphocyte immunophenotyping is an important tool in the diagnosis of immunologic and hematologic disorders. This study used a lysed whole blood method for analysis of lymphocyte sub-populations in samples from children born and living in two districts: a highly-contaminated area (Michalovce) and one (Svidnik/Stropkov) with ≈ 2-fold lower environmental PCB levels. The percentages of B-lymphocytes (CD19(+)), activated HLADR(+)CD19(+) cells, and CD8(+) T-lymphocytes significantly increased at 6- and 16-months-of-age in both selected regions as compared to in cord blood values (p < 0.001). Levels of CD3(+) cells increased significantly (from 61 to 65%) in samples from Michalovce (p < 0.01). Levels of CD4(+) T-lymphocytes declined 10% among 16-month-olds in both regions (Michalovce at p < 0.001 and Svidnik/Stropkov at p < 0.01). Natural killer (NK) cell levels decreased 50% in Michalovce 6- and 16-month-old children and 42% among 6-month-olds in Svidnik/Stropkov (p < 0.001). Compared with the less-contaminated region, Michalovce samples showed significantly higher expression of CD3(+) T-lymphocytes, B-lymphocytes, and activated B-lymphocytes, whereas NK cells were less expressed. Even after adjustment for selected covariates, e.g., maternal cigarette smoking, age, parity, ethnicity, birth weight, and gender of infant, the levels of CD19(+), HLADR(+)CD19(+), and CD3(-)CD(16 + 56)(+) cells were seen to remain significantly different between the districts. These results showed that early-life environmental PCB exposure was associated with fluctuations in major lymphocyte subsets in children, suggesting that there is a post-natal immune system response to PCB exposures.

The relationship between cell surface markers, cytokines, ageing, and cigarette smoking.

The purpose of this study was to investigate the modulation of selected cell surface markers and proinflammatory cytokines production in relation to ageing, and cigarette smoking. The analysis of cell surface receptors was performed by the flow cytometry and cytokines levels were evaluated by the sandwich enzyme immunoassays. We found a decreased expression of CD69, CD28, CD11b, CD95 markers in old population compared to young people (p<0.05; p<0.001). The memory CD45RO lymphocytes were markedly expanded in older population in comparison to young donors (12.93+/-5.92 %, p<0.001) and the selectin CD62L was significantly increased on granulocytes in aged people (p<0.05). Our findings demonstrated an augmented level of CD3 and CD28 on lymphocytes in smokers (p<0.05; p<0.005). The significant depression of CD16+56 molecule was recorded in smokers (10.86+/-0.80%) when compared to non-smokers (14.44+/-0.46; p<0.05). Our results showed a significantly diminished levels of interleukin (IL)-1beta (1.93+/-0.48 pg/ml), and increased levels of IL-6 and tumor necrosis factor (TNF)-alpha in elderly population compared to young people (p<0.05; p<0.001). The present data support previous suggestions that senescence and cigarette smoking may contribute to changes in the immune system activity, resulting in altered cell surface marker expression and cytokine levels (Tab. 1, Fig. 3, Ref. 81). Full Text (Free, PDF) www.bmj.sk.

Hyperacute rejection of living related kidney grafts caused by endothelial cell-specific antibodies: case reports.

We describe two cases of hyperacute humoral rejection of living related kidney grafts despite negative pretransplantation T- and B-lymphocyte flow cytometric crossmatches and blood group identity. Retrospectively, antiendothelial IgG antibodies were detected on a panel of umbilical cord cells in the first case, and IgM antibodies against donor endothelial precursor cells were detected using a new endothelial cell crossmatch kit in the second case. Standard crossmatch methods using donor lymphocytes failed to detect these pathogenic antibodies and did not predict the danger of hyperacute rejection.

Long-term (56-week) oral administration of probiotic Enterococcus faecium M-74 decreases the expression of sICAM-1 and monocyte CD54, and increases that of lymphocyte CD49d in humans.

OBJECTIVES: To study the effects of long-term probiotic [Enterococcus faecium (EF) M-74 strain] application in humans with respect to adhesion molecules, both soluble forms (sICAM-1, sPECAM-1) and their expression on leukocytes. METHODS: Double-blinded randomized and placebo controlled study lasting for 60 weeks. A single capsule containing either 2x10(9) of bacteria EF M-74 with 50 microg of organically bound selenium (E-group) or placebo (P-group) was given to volunteers. Peripheral blood was analyzed for the expression of particular adhesive molecules. RESULTS AND CONCLUSIONS: We observed significant changes in CAMs expression in terms of a decrease in sICAM-1, CD54 on monocytes and CD11b on lymphocytes after one-year administration of Enterococcus faecium M-74 in humans. Anti-adhesion-aimed therapeutic modalities may provide the future approach to prevention and treatment of cardiovascular diseases. Application of probiotics may be part of such strategies. (Tab. 2, Fig. 6, Ref. 41.)

DNA repair and cyclin D1 polymorphisms and styrene-induced genotoxicity and immunotoxicity.

1-SO-adenine DNA adducts, DNA single-strand breaks (SBs), chromosomal aberrations (CAs), mutant frequency (MF) at the HPRT gene, and immune parameters (hematological and of humoral immunity) were studied in styrene-exposed human subjects and controls. Results were correlated with genetic polymorphisms in DNA repair genes (XPD, exon 23, XPG, exon 15, XPC, exon 15, XRCC1, exon 10, XRCC3, exon 7) and cell cycle gene cyclin D1. Results for biomarkers of genotoxicity after stratification for the different DNA repair genetic polymorphisms showed that the polymorphism in exon 23 of the XPD gene modulates levels of chromosomal and DNA damage, HPRT MF, and moderately affects DNA adduct levels. The highest levels of biomarkers were associated with the wild-type homozygous AA genotype. The exposed individuals with the wild-type GG genotype for XRCC1 gene exhibited the lowest CA frequencies, compared to those with an A allele (P < 0.05). Cyclin D1 polymorphism seems to modulate the number of leukocytes and lymphocytes in the analyzed subjects. The number of eosinophiles was positively associated with XPD variant C allele and negatively with XRCC1 variant A allele (P < 0.05) and XPC variant C allele (P < 0.05). Immunoglobulin IgA was positively associated with an XRCC3 variant T allele (P < 0.01) and negatively with XPC variant C allele (P < 0.05). Both C3- and C4-complement components were lower in individuals with XRCC3 CT (P < 0.05) and TT genotypes (P < 0.01). Adhesion molecules sL-selectin and sICAM-1 were associated with XPC genotype (P < 0.05). Individual susceptibility may be reflected in genotoxic and immunotoxic responses to environmental and occupational exposures to xenobiotics.

Simultaneous flow cytometric evaluation of phagocytosis and oxidative burst in human polymorphonuclear cells.

Phagocytosis and oxidative burst (OXIBURST) activity of human polymorphonuclear cells (PMNs) has been simultaneously measured directly in whole blood samples. The ingestion of yeast was assessed by the phagocytosis activity (FA) and phagocytosis index (FI), and the respiratory burst of PMNs was determined as dihydroethidine (DHE) oxidation. We received comparable results in the ingestion of yeast cells by PMNs using either light microscopy (77.31+/-7.56) or flow cytometry detection method (78.26+/-5.14). The significant differences (p<0.05) in FI and OXIBURST activity were find in the patients (2.29+/-0.29 and 14.67+/-3.99, respectively) when compared to healthy donors (1.64+/-0.21 and 32.38+/-14.94, respectively). The two-color flow cytometric procedure permits measurement of two different functions of neutrophils in one step. This flow cytometric procedure is simple, rapid and has the potential to be an alternative assay to test leukocyte function. (Fig. 3, Ref: 30.)

One-year application of probiotic strain Enterococcus faecium M-74 decreases serum cholesterol levels.

OBJECTIVES: To investigate the impact of long-term orally administered probiotic strain Enterococcus faecium (EF) M-74 enriched with selenium on lipid profile (total cholesterol, LDH, HDL, and triglycerides) in humans. BACKGROUND: The discovery that hypercholesterolemia plays a major role in the development of atherosclerosis has led to a number of pharmacological and non-pharmacological (including dietary) approaches resulting in its elimination. Up to now, the question of supposed hypocholesterolemic effect of probiotics has not been definitely established. METHODS: 43 volunteers were randomized into two groups. Participants were given single capsule a day containing 2x10(9) of EF M-74 plus 50 microg of organically bound selenium (E-group) or placebo (P-group). The study was double-blind and lasted 60 weeks. Peripheral blood was analyzed for lipid parameters before intervention, after 6, 12, 23, 44, and 56 weeks of capsule administration, and four weeks following interruption of administration. RESULTS: After 56 weeks of application, decrease in total cholesterol in E-group (17/3 women/men, mean age 75.4+/-1.5 year) was observed (5.94+/-0.29 mmol/l at week 0 vs 5.22+/-0.25 mmol/l after 56 weeks, p<0.001). This reduction was achieved mainly due to a fall in LDL cholesterol (3.85+/-0.27 vs 3.09+/-0.21 mmol/l, p<0.001), as no significant alterations in HDL and triglycerides were noted. In placebo group (14/4, 78.1+/-1.7 year), no statistically important changes were observed after one-year capsule administration. CONCLUSIONS: In our study, the administration of E. faecium M-74 probiotic strain was associated with reduction of serum cholesterol concentration by 12% after 56 weeks. The crescent amount of facts on this issue gives a solid reason to assume that probiotics will find their place as a therapeutic alternative in human medicine. (Tab. 4, Fig. 4, Ref: 36.)

Different patterns of serum interleukin 10 response to treatment with anti-tumor necrosis factor alpha antibody (infliximab) in Crohn's disease.

Administration of anti-tumor necrosis factor antibody (anti-TNF, infliximab) down-regulates T helper 1 (Th 1) cytokines production in intestinal mucosa of patients with Crohn's disease (CD). Interleukin 10 (IL-10) is thought to be involved in CD pathogenesis through regulation of the Th 1 response. The aim of this study was to determine the IL-10 response in CD patients treated with anti-TNF. Fourteen patients with active CD received 5 mg/kg of infliximab; clinical activity assessed by Crohn's Disease Activity Index (CDAI), alpha1-acid glycoprotein and serum IL-10 were determined before and after treatment, in month 0, 1 and 5. In the group with a good clinical response, IL-10 levels diminished significantly in month 1 (p<0.05) and remained decreased in month 5. The group with a lower response showed a significant increase in IL-10 levels in month 1 (p<0.05). alpha1-acid glycoprotein levels obtained before treatment were significantly elevated in the group with a good clinical response (p<0.05) and a significant decrease in month 1 was observed in this group (p<0.05). These observations suggest that a pattern of IL-10 response might be related to the clinical response to anti-TNF treatment in CD.

Antiasthmatic effects of nedocromil sodium.

Accumulating data indicate that bronchial asthma is a chronic inflammatory disease. Airway inflammation and it's control became a principal focus in asthma treatment. Nedocromil sodium is chemically nonsteroidal anti-inflammatory agent for the treatment of mild to moderate asthma. The aim of the study was to determine the effects of NS on bronchial hyperresponsivness and eosinophil activation markers isolated from peripheral blood of asthmatics with mild intermittent asthma. Twenty nine patients of both sexes (17 women, 12 men) with average age of 34 years were recruited into the clinical open study. Bronchial responsivness was assessed by metacholine challenge test prior to starting therapy with NS (preparation Tilade mint aer) and 3rd week and 9th week of follow up. Baseline lung function tests were performed at intervals before treatment and at 3rd and 9th week, respectively. Eosinophil activation markers were determined before and after 3rd and 6th week. Assessement was done by flow cytometry using standard monoclonal antibodies. Bronchial responsivness decreased significantly at 3rd and 9th week of follow up (provocation dose--PD20 increased significantly, p < 0.05, p < 0.02, respectively). Improvements of baseline lung function tests were observed in majority of parameters: FVC (p < 0.01), FEV1 (p < 0.01), FEV1/FVC (p < 0.01), MEF 25 (p < 0.03), MEF 50 (p < 0.01), MEF 25-75 (p < 0.01), PEF (p < 0.01) after 3rd week, however the enhancement of improvement was seen in majority of parameters after 9th week of the study. Significant reduction of eosinophil activation markers expression was noticed: CD69 (p < 0.05, p < 0.01) and HLA DR (p < 0.05, p < 0.05) after 3rd and 6th week, respectively and CD66 (p < 0.05) after 3rd week and CD81 (p < 0.05) after 6th week of follow up. NS possessed complex antiasthmatic effects resulting in decrease of bronchial responsivness and reduction of eosinophil activation markers in mild asthmatics. The tolerance of the drug was good and no adverse effects have been reported. NS is effective prophylactic drug recommended for use in both adults and children in long-term management of mild asthma. (Tab. 2, Fig. 1, Ref. 27).

Decreased levels of coenzyme Q(10) in patients with bronchial asthma.

BACKGROUND: The contribution of free oxygen radicals in the pathogenesis of bronchial asthma is generally accepted. The modulation of antioxidative defence by supplementation with antioxidants represents additive therapy in complex management of disease. The aim of the study was to assess the levels of coenzyme Q10, alpha-tocopherol, and beta-carotene both in plasma and whole blood, and malondialdehyde (MDA) and eosinophil cationic protein (ECP) in plasma of asthmatics (As). METHODS: Fifty-six As (15 males and 41 females) aged from 19 to 72 years (mean age 46 years) suffering from allergic asthma were enrolled into the study. The control group comprised 25 healthy volunteers (16 males, 9 females) aged 25-50 years. RESULTS: The concentrations of CoQ10 decreased significantly both in plasma and whole blood, compared with healthy volunteers (0.34 +/- 0.15 micromol/l vs. 0.52 +/- 0.15 micromol/l, 0.33 +/- 0.14 micromol/l vs. 0.50 +/- 0.13 micromol/l, P < 0.001, P< 0.001, respectively). The levels of alpha-tocopherol were decreased both in plasma and whole blood in comparison with controls [24.10 micromol/l (19.8; 30.5), vs. 33.20 micromol/l (28.25; 38.05), 17.22 +/- 6.45 micromol/l vs. 21.58 +/- 7.92 micromol/l, P= 0.006, P = 0.01, respectively]. The levels of MDA were elevated over the reference range in both groups (reference range < 4.5 micromol/l). No changes were seen in beta-carotene concentrations. Positive correlation was found between whole blood CoQ10 and alpha-tocopherol concentrations. CONCLUSION: Results of the study suggest a possible contribution of suboptimal concentrations of CoQ10 on antioxidative dysbalance in As and provide a rationale for its supplementation.

Effects of occupational exposure to styrene on expression of adhesion molecule on leukocytes.

Styrene is an indispensable chemical extensively used in plastic and synthetic rubber industries. Styrene is known to produce various types of hepatotoxic, neurotoxic, and genotoxic effects. Styrene may be immunotoxic by both direct and indirect mechanisms. Measurement of adhesion molecules is a new tool for the investigation of immune system modulation. The aim of this study was to evaluate the association of the expression of the adhesion molecules CD11a, CD11b, CD18, CD54, CD49d, and CD62-L in white blood cells and levels of soluble adhesion molecules ICAM-1 and L-selectin in serum with occupational exposure to styrene. Analyses by flow cytometry revealed elevated levels of most of the assessed adhesion molecules on surfaces of lymphocytes, monocytes, and granulocytes. Expression of the adhesion receptor antigens CD11a on lymphocytes, CD11b on monocytes, and CD18 on granulocytes were unaffected. Workers exposed to styrene had decreased concentrations of sICAM-1 and no changes in concentrations of sL-selectin. Styrene exposure appears to increase activation of the immune system and alter leukocyte adherence. This interaction is a critical first step in immune stimulation and leukocyte-endothelial interaction.

Effects of selenium supplementation on expression of adhesion molecules in corticoid-dependent asthmatics.

Selenium (Se) deficiency attenuates the host immune response, thereby increasing the risk of bacterial and viral infections. We have examined the effects of selenium supplementation (SeS) in corticoid-dependent asthmatics (CDAs) with lowered circulatory Se status. Twenty CDAs (10 males and 10 females, average age 54.5 yrs) were enrolled into the study. The average duration of the disease was 10 yrs. The asthmatics were receiving 200 micrograms of Se per day for a period of 6 months, in addition to regular treatment with inhaled corticosteroids and beta-agonists. The expression of adhesion molecules (CD11a, CD11b, CD18, CD49d, CD54, CD62L) on peripheral blood mononuclear cells (PBMCs) of asthmatics and the expression of E- and P-selectins, ICAM-1, VCAM-1 on cultured human umbilical vein endothelial cells (HUVEC) after stimulation with PBMCs from CDAs before and after 3 and 6 months of SeS were assessed by standard monoclonal antibodies and analyzed by flow cytometry. The concentrations of soluble adhesion molecules P-selectin, E-selectin, ICAM-1 and VCAM-1 were determined by ELISA method. The expression of adhesion molecules on PBMCs: After 3- and 6-months of SeS, a decreased expression of molecules CD11a, CD11b and CD62L was observed (p < 0.02, p < 0.005, p < 0.003). No changes were seen in the expression of CD18, CD49d except for the increased expression of CD54 (p < 0.005). Modulation of adhesion molecules expression on HUVEC: We observed a significant increase in VCAM-1, P- and E-selectins expressions in the group of asthmatics without SeS in comparison with the control group (p < 0.05, p < 0.01, p < 0.05). During SeS a significant decrease in molecules VCAM-1, E-selectin (after 3 months) (p < 0.05, p < 0.05) and P-selectins and ICAM-1 (after 6 months) (p < 0.05, p < 0.01) were observed. Soluble adhesion molecules: After 3 months of SeS we noticed a significant decrease in VCAM-1 and P-selectin expressions (p < 0.05, p < 0.05) and after 6 months the level of VCAM-1 decreased (p < 0.01). The effect of Se on the adhesion molecules expression in endothelial cells in vitro experiments: Se blocks the expression of adhesion molecules stimulated by IFN-gamma in a dose-dependent way after addition of Se into a culture of endothelial cells. Concentration of 10 micrograms/ml inhibits the increase in expression of ICAM-1 (p < 0.05) but not that of VCAM-1, E- or P-selectins. The inhibition of expression in Se concentration of 10 micrograms/ml is over 80% (p < 0.01). Our data demonstrate that Se is able to affect the adhesion molecules expressions that are crucial in the inflammatory process. (Fig. 5, Ref. 22.)

The influence of selenium supplementation on the immunity of corticoid-dependent asthmatics.

UNLABELLED: Selenium (Se) is a trace element that is essential for immune functions, and protects the immune system from oxidative damage. AIM: The aim of the pilot clinical study was to assess the influence of selenium supplementation (SeS) on the selected immune parameters analyzed from peripheral blood of corticoid-dependent asthmatics (CDAs). MATERIAL AND METHODS: Seventeen CDAs aged from 30 to 74 years (7 females, 10 males) with suboptimal levels of Se in plasma were enrolled into the study. The follow up of SeS lasted 96 weeks. It is daily dose was 200 micrograms (2 x 2 tbl daily, 1 tbl contained 50 micrograms of Se). Before (-4 weeks) and after the 12th, 48th, 72nd and 96th weeks of SeS, the following parameters were observed: Epitopes EG1, EG2 expressed on intracellular eosinophil (Eo) cationic protein and eosinophil peroxidase, the numbers of CD3, CD4, CD8, CD19 and CD3 HLADR positive T lymphocytes (Ly), lymphocyte blastogenesis test (LTT) with mitogens concanavalin A, (Conc A) phytohemaglutin (PHA), the levels of C3, C4 complement components, activation of complement by classic and alternative pathways (CP50, AP50), the levels of immunoglobulins (Ig) G, A, M and total IgE, circulating immune complexes (CIC). RESULTS: Epitopes EG1 and EG2 in cytoplasma of Eo decreased significantly after 12 weeks of SeS, (p < 0.01) and 96 weeks of follow up. In parameters of T cell mediated immunity the relative number of CD3 HLADR+ T Ly increased after 24, 48 and 96 weeks of SeS (p < 0.0008, p < 0.009, p < 0.07). Proliferative activity of T Ly to mitogenes PHA and ConcA in LTT decreased significantly after 12, 48, 72 and 96 weeks of SeS (p < 0.0005, p < 0.009, p < 0.04, p < 0.02, respectively). In humoral parameters activation of CP50 decreased after 24, 72 and 96 weeks of SeS to the reference range (p < 0.001, p < 0.03, p < 0.02) and AP50 after 96 weeks, respectively (p < 0.02). The levels of IgG elevated after 24 weeks (p < 0.02), IgA after 24, 48 weeks (p < 0.0007, p < 0.02, respectively). The level of total IgE significantly decreased after 96 weeks of SeS (p < 0.003). CONCLUSION: Our pilot clinical study with the CDAs demonstrates the significant changes particularly in functional parameters of both cellular and humoral types of immunity. These results support the immunomodulating effects of SeS. (Tab. 5, Ref. 15.)

Detection of antiendothelial cell antibodies in patients with connective tissue diseases by flow cytometry and their relation to endothelial cell activation.

Antiendothelial cell antibodies (AECA) have been detected by flow cytometry analysis in 23 out of 80 patients with connective tissue diseases. Ten out of 19 serum samples from patients with systemic lupus erythematosus (SLE) were positive. These antibodies were not detectable in healthy donors. We examined the capacity of serum samples to induce endothelial cell activation by modulating cell adhesion molecule expression on human umbilical vein endothelial cells. We found that sera from both AECA-positive and AECA-negative patient groups induced a significantly higher expression of E-selectin compared to healthy controls (P<0.05). There were no differences in the ICAM-1 on VCAM-1 expression. Our data suggest that increased E-selectin expression in activated endothelium in patients with various connective tissue disorders is not related to the production of AECA.

Inflammatory mechanisms involving neutrophils in chronic venous insufficiency of lower limbs.

BACKGROUND: It is supposed that an inflammatory reaction is one of the major factors responsible for the chronic venous insufficiency (CVI) of lower limbs which cause leg ulcers. OBJECTIVES: The main objective of the present study was to determine the differences in the levels of typical inflammatory mediators and markers produced by neutrophils of patients with CVI and normal control subjects. SUBJECTS AND METHODS: 26 patients with CVI and 39 clinically healthy subjects were included in the study. In peripheral neutrophils of both groups the production of superoxide, total reactive oxygen intermediates and activities of lysosomal enzymes were measured together with the expression of 8 adhesion molecules. RESULTS: Increased formation of superoxide by patient neutrophils and activities of elastase in both neutrophils and serum of patients were demonstrated. On the contrary, activities of myeloperoxidase and beta-D-glucuronidase were decreased in patient neutrophils. Comparing to control group adhesion molecules CD11b, CD18, CD31, CD49d, CD54 and CD62L were increased on the surface of patient neutrophils whereas no differences were observed in the expression of CD11a abd CD15. CONCLUSION: The neutrophils of patients with CVI are primed and/or activated because they are able to release higher amount of superoxide, lysosomal enzymes and express elevated number of adhesion molecules. It may serve as one of the important evidences of an inflammatory mechanism involved in the pathogenesis of chronic venous insufficiency. (Tab. 3, Ref. 27.)

Detection of intracellular cytokines during antioxidant supplementation in corticoid-dependent asthmatics and modulation of adhesion molecule expression on cultured endothelial cells.

In this study, we report on the interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) cytokine responses to phorbol myristate acetate (PMA) + ionomycin-stimulated CD3+ lymphocytes in asthmatic subjects when compared with normal donors. There was a significantly lower production of intracellular IFN-gamma in asthmatic patients. No difference was found for IL-4 production between these two groups. After administration of a multivitamin-mineral supplement containing selenium, zinc, vitamin A, vitamin B6, vitamin C, and vitamin E for 6 mo, a significant increase in the percentage of CD3+/IL-4 positive cells (p < 0.05) was found. The induction of endothelial cell adhesion molecule (CAM) expression in cultured human umbilical vein endothelial cells (HUVEC) and whole-blood mixture was studied using flow cytometry. The ICAM-1 and VCAM-1 expressions were higher in the patients than in control donors (p < 0.05). There is a correlation between the increased percentage of CD3+/IFN-gamma positive cells and reduced endothelial ICAM-1 and VCAM-1 expression after 6 mo of intervention period. No apparent effect of supplementation on CAM expression was found, suggesting that these changes do not arise from an antioxidant mechanism. This newly developed whole-blood technique for the assessment of CAM expression can be of use for monitoring therapy in inflammatory diseases.

Significance of anti-interferon-alpha2 and sICAM-1 activities in the sera of viral hepatitis B and C patients treated with human recombinant interferon-alpha2.

In this study the presence of an IFN-binding activity in the sera of patients with chronic viral hepatitis B or C treated with rIFN-alpha2 was screened by a radioimmune assay (RIA) using radiolabeled rIFN-alpha2. Incidence of an anti-IFN activitywas compared with hepatitis B virus (HBV) or hepatitis C virus (HCV) serum markers as hepatitis B s antigen (HBsAg), hepatitis B e antigen (HBeAg), antibodies to HBsAg (anti-HBsAg), antibodies to HBeAg (anti-HBeAg), seroconversion, HBV DNA, HCV RNA, and serum soluble intracellular adhesion molecule I (sICAM). Injections (intramuscular) of rIFN-alpha2 caused an anti-rIFN activity formation in 8 (27.6%) of 29 patients with chronic active hepatitis B (CAH-B) and in 8 (30.8%) of 26 patients with chronic active hepatitis C (CAH-C). The presence of the anti-rIFN activity in CAH-B patients correlated frequently with the persistence of HBsAg, HBeAg and HBV-DNA, while its absence was often accompanied by the anti-HBeAg and anti-HBsAg seroconversion, respectively, and HBV-DNA negativity. In two CAH-C patients who became HCV RNA-negative no anti-IFN activity was found. Levels of serum sICAM-1 in CAH-B patients responding to the IFN treatment were higher than those in non-responders or in which the anti-IFN activity was present. The anti-IFN activity may negatively influence the effect of the IFN therapy of CAH-B or CAH-C patients at early stages of the therapy. The appearance of the anti-IFN activity at the end of a long-term IFN therapy does not seem to influence the outcome of the therapy. sICAM-1 may be involved in the process of CAH-B reactivation and IFN-triggered cytotoxicity during the IFN therapy.

[The vascular endothelium in inflammatory processes in vivo and in vitro]. / Cievny endotel v zápalovom procese in vivo a in vitro.

Endothelial injury occurs in atherosclerosis, infectious, rheumatic and vasculitic processes, leading to activation of transcription factors and endothelial expression of various cytokines and adhesion molecules. Endothelial cell cultures represent a valuable tool in research activities, with emphasis on the principal characteristics of angiogenesis, inflammatory response, transduction signals and endothelial functionality. In the laboratory of tissue cultures we prepared primary endothelial cultures by their isolation from the umbilical vein. This model system has been used to investigate the endothelial activation in vitro, adhesion alterations of immunocompetent cells to endothelium, adhesion molecule expression in the disease course monitoring and anti-inflammatory treatment. (Tab. 1, Fig. 5, Ref. 45.)