Characteristic of circulating immune complexes in the blood of children with asthma

Circulating immune complexes (CIC), which are heterogeneous in size and composition population of antibodies and antigens, are critical mediators of the immune system. Precipitation of the CIC on the vascular wall and basal cell membranes causes a variety of biological effects: activation of the complement system in accordance with the classical pathway, activation of macrophages initiating the development of a local inflammatory process involving mast cells and neutrophils. In the present study, we have examined the content and physical characteristics of the CIC in the serum of children with atopic asthma and healthy individuals. We have found that the concentration of CIC in patients is 2-fold higher than in control group. The presence of the most pathogenic medium-sized complexes was determined in 9 % of healthy children and 85.6 % of children with asthma

Electron immunohistochemical analysis of localization of neutral Mn2+-dependent DNAase. II. Analysis of ultrastructural localization in epon sections of different organs of the rat | Elektronnoe immunogistokhimicheskoe izuchenie lokalizatsii neitral'noi Mn2+-zavisimoi DNKazy. II. Ul'trastrukturnaia lokalizatsiia DNKazy na éponovykh srezakh razlichnykh organov krysy.

The use of a conjugate of colloidal gold with monotypic antibodies against a neutral Mn(2+)-dependent DNAse has revealed the enzyme localization in ultrathin Epon sections of glutaraldehyde fixed tissues. Technical procedures involved in fixation, embedding, and immune reactions are described. DNAse has been established in the nuclei in both normal and regenerating liver. A protein, immunologically close to DNAse, has been identified in the nuclei of a lymph node, thymus, spleen and cerebellar cortex

Electron immunohistochemical analysis of localization of neutral Mn2+-dependent DNAase. III. Visualization of DNAse binding to isolated chromatin | Elektronnoe immunogistokhimicheskoe izuchenie lokalizatsii neitral'noi Mn2+-zavisimoi DNKazy. III. Vizualizatsiia sviazyvaniia DNKazy s izolirovannym khromatinom.

Neutral Mn(2+)-dependent DNAse is localized on isolated chromatin structures in both normal and regenerating rat liver. The enzyme was revealed located along the whole length of nucleosomal chain and in hypernucleosomal structures. However, as concerns the quantity of the enzyme, it was distributed unevently along the chromatin, thus reflecting the pattern of different functional states of native chromatin. According to biochemical and immunohistochemical data, DNAse can hydrolyse in vitro only one-stranded DNA. One of possible explanations of the observed differences in DNAse binding with native DNA chromatin and its inability to adsorb on native DNA in vitro may be the presence of hypothetical DNA-binding proteins in native chromatin making complexes with DNAse and thereby responsible for immobilization of the enzyme on chromatin structures in vivo

Electron immunohistochemical analysis of localization of neutral Mn2+-dependent DNAase. I. Synthesis of ferritin and colloidal gold conjugates with monospecific antibodies against neutral Mn2+-dependent DNAase | Elektronnoe immunogistokhimicheskoe izuchenie lokalizatsii neitral'noi Mn2+-zavisimoi DNKazy. I. Sintez kon''iugatov monospetsificheskikh antitel k neitral'noi Mn2+-zavisimoi DNKaze s ferritinom i kolloidnym zolotom.

Rabbit antibodies against a neutral Mn(2+)-dependent rat liver DNAse were obtained, whose specificity towards DNAse was ascertained by suppression of the enzyme activity both in vitro system and immunoblotting assays. Procedures of synthesis of ferritin and colloidal gold conjugates with antibodies are described. The biological activity of the conjugates proved to be similar to that of the original antibodies

The origin of elevated levels of circulating DNA in blood plasma of premature neonates

The relationship between the concentration of cell-free DNA (cfDNA) and the number of proliferating/apoptotic lymphocytes in peripheral blood of premature newborns of different gestation age and full-term newborns was determined. The experiments were performed using fluorescent spectrophotometry (with Hoechst 33342), flow cytometry, and microscopy (Feulgen staining of lymphocytes). It was determined that the lymphocyte population of premature newborns may consist of 4.6% of proliferating and 22.1% apoptotic cells. For full-term newborns, the percentage was 2.5% and 2.9%, respectively. A direct correlation between the concentration of extracellular DNA and the number of proliferating lymphocytes of full-term newborns was ascertained (r = 0.400; P < 0.05). For premature newborns, the concentration of extracellular DNA correlated both with proliferating lymphocytes and apoptotic cells. The results show that premature birth causes the induction in lymphocytes of both apoptosis and proliferation that are accompanied by an increased extracellular DNA concentration in the blood of newborn babies. © 2008 New York Academy of Sciences

Dexamethasone affect on the expression of bcl-2 and mTOR genes in T-lymphocytes from healthy donors

Synthetic glucocorticoids are able to activate apoptosis in the cells by regulating the transcription of the respective genes. Effect of dexamethasone on apoptosis is an established fact. However, its influence on another program of cell death autophagy, is currently unproven. Therefore, in this paper we have analyzed the influence of dexamethasone on the expression of bcl-2 and mTOR genes in T-lymphocytes from healthy donors. The results showed that dexamethasone reduced the expression of bcl-2 and mTOR genes. However, the nature of the effect of dexamethasone on mTOR and bcl-2 expression was different: the expression of bcl-2 gene in the long-term cultivation was maintained at the same reduced level, while the expression of mTOR was first reduced and then increased

Autophagy induction in peripheral blood T-lymphocytes of atopic asthma patients

Autophagy is a fundamental process that ensures the regulation of T-cell homeostasis. In case of apoptosis induction disruption in the cell it could be single mechanism of the cell death. Previously was shown inhibition of lymphocyte apoptosis in patients with bronchial asthma, so the main study of this work has focused on the study development process of autophagy in T-lymphocytes of patients with bronchial asthma. The article presents the main morphological changes in cells associated with activation of autophagy (formation autophagosome). In addition to morphological changes in lymphocytes, we have shown the expression of autophagy marker protein (LC3B). We found that in T-lymphocytes of patients with severe asthma are simultaneous activation of both autophagy and apoptosis, and autophagy is a stimulus to cell death