[In vitro migration activity of human peripheral blood neutrophils in the normal state and in immunopathology]. / Migratsionnaia aktivnost' in vitro neitrofilov perifericheskoi krovi cheloveka v norme i pri immunopatologii.

Methodological approaches to evaluation of the migration activity of human peripheral blood neutrophils into a collagen matrix were worked out. The migration of neutrophils in healthy donors and in patients with severe bronchial asthma was studied. In the normal state there was practically no migration of intact neutrophils into the collagen matrix (1.1 +/- 0.4%). Following their stimulation by formyi peptide about a quarter of their population was drawn into the matrix in avalanche (22.0 +/- 5.9%). In the acute phase of severe bronchial asthma an increase in both spontaneous (3.3 +/- 1.5%, P < 0.01) and stimulated (35.6 +/- 4.6%, P < 0.001) cell migration occurred. Changes in the migration characteristics of the neutrophils of patients and those of the cells of healthy donors, treated with the polycytokine preparation at concentrations exceeding 100 g/ml, followed similar trends. In case of the standard asthma treatment along with positive disease dynamics further increase in spontaneous neutrophil migration (5.8 +/- 2.9%, P < 0.001) in combination with deficiency in cells reaction to formyi peptide (11.8 +/- 3.8%, P < 0.01) was registered. At the same time dexamethasone did not change the character of the in vitro migration of neutrophils into the collagen matrix. Thus the dynamics of the peripheral blood neutrophil migration during treatment of severe bronchial asthma was demonstrated; this dynamics could be indicative of the pathogenetic role of neutrophils in the development of this pathology.

[Level of serum antibodies to opportunistic microflora as a marker of the secondary immunodeficiency processes in children]. / Uroven' syvorotochnykh antitel k uslovnopatogennoi mikroflore kak marker protsessa formirovaniia vtorichnogo immunodefitsita u detei.

Examination of children with different noninfectious diseases resulted in obtaining the data base on the state of health of 201 children belonging to the potential risk group of the development of secondary immunodeficiency. The children were subdivided into several groups which differed by the type of immune disturbances and accompanying metabolic shifts. The level of antibodies to one of the fragments of peptidoglycan-N-acetylmuramyldipeptide was compared with the character of changes in the immune system. Different titers of serum antibodies to peptidoglycan were found to correspond to different forms of immune disturbances. The study showed that from the group with the absence of definite signs of immunodeficiency to the group with the pronounced deficiency of T lymphocytes and monocytes expressing CD14 antigen changes in the immune system increased in parallel to a rise in the concentration of alpha 2-macroglobulin, the development of hypercholesterolemia and a decreased level of antibodies to peptidoglycan. Opportunistic microflora was seemingly an important factor in the formation of definite forms of disturbances of the immune system and accompanying metabolic shifts.

[Pathogenetic role of intestinal microbiocenosis in the pathogenesis of hereditary myodystrophy]. / Patogeneticheskaia rol' kishechnogo mikrobiotsenoza v klinike nasledstvennykh miodistrofii.

The effect of the inclusion of probiotic preparations for the correction of disturbances in normal intestinal microflora into the complex therapy of patients wish Duchenne's childhood muscular dystrophy and Becker's myopathy was analyzed. Probiotic therapy made it possible to improve the clinical state of patients, manifested by an increase in muscular strength and accompanied by positive shifts in electromyographic, immunological, biochemical, hormonal characteristics. Intestinal microbiocenosis plays seemingly a certain role in the formation of hereditary pathology.

[Role of dysbacteriosis in the formation of chronic noninfectious pathology in children]. / Rol' disbakterioza v formirovanii khronicheskoi neinfektsionnoi patologii u detei.

The results of clinical observations and laboratory data make it possible to regard dysbacteriosis as an important factor in the pathogenesis of chronic noninfectious pathology in children. The adequate complex correction of intestinal dysbacteriosis on the basis of probiotic therapy facilitates the prolonged remission of the disease in children with diabetes mellitus of type 1 (DM1) and myopathy, decreases severity of late complications of DM1. A suggestion is made on the role of dysbiotic microflora in the development of chronic non-infectious pathology in children.

[Immunological prognosis of the effectiveness of soybean diet]. / Immunologicheskii prognoz éffektivnosti soevogo pitaniia.

During 50 days a group of 23 volunteers received lunches prepared on the basis of textured proteins of soy beans. Each lunch included 50 g of proteins. Laboratory investigation was carried on dynamic values of lipid metabolism, cell-mediated, humoral and local immunity. A total of more than 30 tests were made. The analysis of immunological reactivity in volunteers has revealed the immunomodulatory activity of soy bean protein similar to the reference modulator with known quantitative estimation of effects on cell mediated, humoral and local immunity. Statistic analysis of preliminary individual laboratory results and plotting of prognostic curves allowed to develop an original method of volunteer selection for whom soy bean diet may bring positive or negative results.

[The apoptogenic mechanisms for the occurrence of immunodeficient diseases]. / Apoptogennye mekhanizmy vozniknoveniia immunodefitsitnykh zabolevanii.

Suggestion has been made that the development of immunodeficient states should be considered as the result of the acceleration of the apoptosis of immune cells. The pathogenetic basis of the apoptogenic mechanism of the development of immunodeficient morbidity is considered.

Dynamics and Mechanisms of Interaction of Mouse Spleen Lymphocytes with 3D Collagen Matrix in vitro.

Interaction of immunocompetent cells with extracellular matrix is one of the main stages in their homing and circulation. In this connection we investigated quantitative and dynamic parameters of interaction between splenocytes and 3D collagen matrix in vitro. It was found out that, about 20% of mouse spleen lymphocytes exhibited ability to bind to type I collagen that reflected as their adhesion to and/or migration in collagen matrix. The number of lymphocytes capable of the interaction with collagen gained successively as far as the time of their incubation on collagen matrix was increased and reached maximum by 24 h. The lymphocyte-collagen interaction was energy-dependent and engaged collagen receptors, which probably have been already expressed on cells before spleen lymphocytes were isolated. The series of intracellular interchanges as activation of protein kinase C, assembly of actin filaments and depolymerization of tubulin microtubules were critical for lymphocytes to adhere to and further to migrate in collagen matrix. Long lasting incubation (24 h and more) of lymphocytes in adhesion excluding conditions did not reduce the number of cells able to interact with collagen, but to a great extent changed mechanisms providing their adhesion and/or migration.

Contact Effects of Mononuclear Cells on L929 Fibroblast Morphological Phenotype.

We studied the contact interactions of human peripheral blood mononuclear cells (MNC) and transformed mouse fibroblast cell line L929 (L-cells), namely their effects on morphological phenotype of L-cells. The morphological characteristics of the fibroblast, (cell area, nucleus-cytoplasmic ratio, cell spreading, cell shape) were estimated with the aid of fight scanning microscopy, followed by computer image analysis. Contact interaction between fibroblasts and MNC caused normalization of morphological phenotype of the fibroblasts (increase of cell area, shape factor, spreading and decrease of nucleus-cytoplasmic ratio). This phenomenon was revealed by analysis of both average morphological characteristics and population contents. Only a particular subpopulation of L-cells, but not the whole population, was shown to be normalized by the effects of MNC. For elucidation of responsible MNC population, which was capable of influencing on fibroblast morphological phenotype, we separated MNC in several cell types: adherent cells, non-adherent cells, which were separated in E-rosetting cells, and non-E-rosetting cells. Only non-adherent and E-rosetting cells could normalize the morphological phenotype. E-rosetting population consisted of 85% of CD3(+) (T lymphocytes) and 15percnt; of CD56(+)/CD16(+) (natural killers). Supernatants of MNC, and MNC cocultured L-cells, obtained after 24 h incubation in the cell culture medium, were not able to normalize the morphological phenotype of the fibroblasts. They had small denormalizing effect on the fibroblasts (decrease of cell area, shape factor, spreading and increase of nucleus-cytoplasmic ratio). The results of this study indicate that the contact interaction between MNC and fibroblasts may normalize transformed fibroblast morphological phenotype. The dependence of fibroblast functional state on their morphological phenotype imply the presence of regulatory mechanism of contact interaction between fibroblasts and MNC, which determines many live processes in fibroblast.

[A model for the assessment of fibroblast morphology]. / Model' dlia otsenki morfologii fibroblastov.

A model for comprehensive assessment of morphologic parameters of a cell (area, nucleus and cytoplasm area, degree of spreading), based on the use of a scanning device, has been developed. The model comprises an original method for staining the preparations, which permits assessment of the degree of cell spreading by microdensitometry, and updating the equipment and software. The programs help assess the integral and derivative parameters, such as the area of the interface zone of a cell, the shape factor, spreading, and nucleus to cytoplasm ratio. Using this model, the effect of peripheral blood mononuclears on the morphology of transformed murine fibroblasts L929 upon a contact exposure was assessed. Addition of mononuclears to fibroblast culture led to alteration of their morphology: increase of the mean cellular area, degree of spreading, and of shape factor and reduction of the nucleus/cytoplasm ratio. This was true for both the mean values and the population composition.

[The interpretation of laboratory parameters in evaluating the immune status of man]. / Interpretatsiia laboratornykh pokazatelei pri otsenke immunnogo statusa cheloveka.

At least 3 physiologic parameters appear to be useful for the assessment of the immunity status of a patient. These parameters are as follows: immunocyte ability to activation, proliferation, and differentiation; these characteristics may be directly examined with the use of novel products of immunobiotechnology. The suggested approach essentially simplifies interpretation of the findings obtained in studies of human immunity system. Besides lymphocytes, special attention is paid to immunoregulatory monocytes, macrophages, and granulocytes. Since these cells are known to secrete quite a number of mediators, an imbalance of their production and reception may contribute to the pathogenesis of many a disease.