[The immunity during the pre-clinical period of type I diabetes mellitus].

The purpose of the study was to investigate the condition of immunity (blood lymphocyte immune phenotype and ultrastructure) in healthy children with a family background of type 1 diabetes mellitus (DM 1) having or not having diabetes-associated autoantibodies (DAAB). The subjects of the study were divided into three groups. Group 1 consisted of 90 children with a family background of DM 1 (first line relatives had DM 1), DAAB- (GADA, IA-2A, and IAA) positive or negative; group 2 consisted of 51 children with newly revealed DM 1; group 3 included 45 healthy controls, normoglycemic DAAB-negative children with no family background of DM 1. GADA, IA-2A, and IAA titers were measured using radioimmunoassay. The immune phenotype of lymphocytes (CD3+, CD4+, CDr8, CD20+, and CD56+ cells) were studied using flow cytometry (FACS-analysis); their ultrastructure was studied by means of electron microscopy. The study found a significantly lower total number of T-lymphocytes (CD3+ cells), T-helpers/inductors (CD4+ cells), and natural killer cells (CD56+ cells and large granule-containing lymphocytes) in the DAAB-positive children vs. the DAAB-negative ones and especially the controls. In the DAAB-positive children, electron microscopy found distinct changes in the ultrastructure of CD4+ lymphocytes and large granule-containing lymphocytes (CD56+ cells), which evidences changes in the secretory and cytostatic function. Such changes in the number and ultrastructure of these lymphocyte subpopulations are found in patients with newly revealed DM 1. Thus, immune changes happen in the organism of a healthy person a long time before clinical manifestations of DM 1 develop; these changes reflect a concealed autoimmune process in Langerhans islets. Detection of DAAB plays a significant role not only in studying poorly understood pre-diabetes nature, but also in the development of new, scientifically based methods of its prevention and treatment.

[The ultrastructure and function of lymphocytes in children with first detected untreated type 1 diabetes mellitus].

Flow cytometry (FACS-analysis), light and electron microscopy, and cytotoxic test were used to study the leukocytic composition, content, ultrastructure, and function of different lymphocytic populations (CD3+, CD4+, CD8+, CD20+, and CD56+ cells) in the blood of 90 children aged 8 to 15 years who had untreated new-onset type 1 diabetes mellitus. A control group consisted of 45 apparently healthy normoglycemic children. The sick children were observed to have a slight, but statistically significant reduction in the relative and absolute blood content of CD3+, CD4+, and CD56+ cells, CD4/CD8 index and particularly large granule-containing lymphocytes (the morpho-logical homologue of natural Idller (NK) cells). Ultrastructurally, CD20+4 cells (lymphocytes containing Gall's bodies) showed the signs of a high functional activity while NK cells (large granule-containing lymphocytes) exhibited a low one, which was confirmed by the decreased cytotoxic activity of these cells determined in vitro. Three-month insulin therapy leading to the restoration of the blood content of glucose and glycosylated hemoglobin fails to normalize the detected changes in the parameters of T- and NK-cell immunity.

[Effect erythropoietin-beta on parameters of erythropoiesis and immunity in patients with diabetes mellitus complicated with diabetic neuropathy and anemia]. / Vliianie éritropoétina-beta na pokazateli éritropoéza i immuniteta u bol'nykh sakharnym diabetom, oslozhnennym diabeticheskoi nefropatiei i anemiei.

Erythropoiesis indices, number of leucocytes in 1 ml of blood, leukocytic formula, lymphocytes immunophenotype in the patients' blood with diabetes mellitus complicated with diabetic neuropathy and marked normochromal anemia after the treatment with Erythropoietin b have been analyzed in the study. Significant increase in haemoglobin, erythrocytes content, hematocrit indices of peripheral blood has been observed during the treatment. The patients felt better, their quality of life improved, CD3+, CD4+, CD5+ cells and granule-contained lymphocytes content normalized. Serious adverse affects of the medication haven't been observed in the study.

[Clinical and immunological studies of the therapeutical effect of cytokines combined with 5-fluorouracil in metastatic kidney cancer]. / Kliniko-immunologicheskoe izuchenie lechebnogo vozdeistviia tsitokinov v sochetanii s 5-ftoruratsilom pri metastaziruiushchem rake pochki.

The authors present the results of a six-year clinicoimmunological study of a therapeutic effect of cytkins combined with 5-fluoruracyl in metastatic renal-cell carcinoma. Two therapeutic regiments have been used: rhIFN-a + 5-FU and rhIL-2 + RHifn-A + 5-FU. In cytokin-sensitive patients, both therapy protocols vrs conventional therapeutic alternatives (cytostatics, hormones, irradiation) have been shown to increase the frequency of achievement of remission by objective scoring and life span of the patients. There was an improvement in patients on having received the complex with IL-2 but a higher therapeutic effect appeared to be accompanied by substantial side effects. Recommendatory measures well-targeted to those patients with metastatic renal-cell carcinoma who are to be placed on cytokinotherapy are presented together with immunological indices to monitor the treatments administered and prognosis.

[The effect of insulin therapy on the level of natural killer cells of different immunological phenotypes (CD16+, CD56+ and CD57+) in the blood of patients with diabetes mellitus type I]. / Vliianie insulinoterapii na soderzhanie estestvennykh kletok-killerov razlichnogo immunologicheskogo fenotipa (CD16+, CD56+ i CD57+) v krovi bol'nykh sakharnym diabetom I tipa.

Flow cytometry with monoclonal antibody Leu-7 (CD57), Leu-11 (CD56) and Leu-19 (CD56) were used to study the content of different subpopulations of natural killer cells (NK-cells) in the blood of type I diabetes mellitus patients before and after insulin treatment and in healthy people. After a course of insulin therapy most patients showed restoration of the total cell number with antigens on their surface characteristic of NK-cells, especially CD56, that indicates the essential role of hypoinsulin in the depression of the NK-system. At the same time a small group of patients was distinguished who did not show such normalization. This may indicate participation of other mechanisms in the formation of natural immunity failure, in particular, the genetic.

[The number and ultrastructure of CD4+ lymphocytes in the blood of healthy persons and in patients with type 1 diabetes mellitus]. / Soderzhanie u u'ltrastruktura CD4+-limfotsitov krovi zdorovykh liudei i bo'lnykh sakharnym diabetom I tipa.

The flowing cytofluorometric and electron microscopic methods have been used to study the content and ultrastructure of blood CD4 lymphocytes in healthy and type I diabetic patients. Analysis of submicroscopic structure of the lymphocytes expressing CD4 antigen has revealed its morphological heterogeneity. It has permitted distinguishing four groups of cells. A decrease of the content of CD4+ lymphocytes at the expense of the first group cells and relative increase in the number of the fourth-group cells are observed in the majority of diabetic patients. The availability of parallel-tubular structures in granules forming clusters and first-group lymphocytes, an increase in the number and size of granules of the fourth-group are the most typical changes of ultrastructural characteristics.

[Cytofluorimetric analysis of blood lymphocyte subpopulations detected using monoclonal antibodies in patients with type I diabetes mellitus]. / Tsitofliuorimetricheskii analiz subpopuliatsii limfotsitov krovi, vyiavliaemykh monoklonal'nymi antitelami, u bol'nykh sakharnym diabetom I tipa.

The content of lymphocytes of different immunological phenotypes: T11+ (all T-lymphocytes), T8+ (suppressors/killers), T4+ (inducers/helpers) and NKH1+ (natural killer cells) was investigated in the blood of 11 patients with type I diabetes mellitus by the method of flow-rate cytofluorimetry using a laser cell sorting device EPICS-C (Coulter), and a kit of monoclonal antibodies; 16 primary donors entered into the control group. As compared to the controls the patients demonstrated a statistically significant decrease in the absolute content of T11+, T4+, T8+ and especially NKH1+-cells. The results obtained confirmed previous data reported by us and other authors concerning disorder in the system of NK cells in insulin dependent diabetes mellitus. These results accounted for frequent development of infectious-inflammatory complications in this disease.

[Ultrastructure of large granule-containing lymphocytes possessing natural killer activity]. / Ul'trastruktura bol'shikh granulosoderzhashchikh limfotsitov, obladaiushchikh estestvennoi killernoi aktivnost'iu.

By means of light and electron microscopy, ultrastructural cytochemistry and immune cytochemistry methods, contents and ultrastructure of large granule-containing lymphocytes (LGL) have been studied in human blood--this is cell population possessing natural killer and, partly, antibody-depending cytotoxicity. The LGL concentrates are isolated from blood applying successive physical-chemical methods, differential centrifugation in the density gradient of pack-phycoll and percoll included. Separate LGL populations are marked by means of rosette-forming reaction with sheep erythrocytes and monoclonal antibodies OKT4 and OKT8. Relative and absolute amount of the LGL in 1 1 of blood is 5.4 +/- 0.5% and 0.319 +/- 0.28 X 10(9), respectively. The LGL ultrastructure is characterized with a low nuclear-cytoplasmic ratio, with presence of osmiophilic (azurophilic) granules in cytoplasm and specific parallel-tubular structures, with a well developed Golgi complex, an essential number of mitochondria, vesicles with smooth wall and vacuoles, as well as multivesicular bodies and Gallo bodies. The LGL subpopulations, expressing various membrane antigens (E+, E-, OKT8+, OKT8-) differ in their ultrastructure, that is evidently stipulated by the degree of their differentiation and their function.

[Ultrastructure of human blood T-lymphocytes marked with monoclonal antibodies]. / Ul'struktura T-limfotsitov krovi cheloveka, markirovannykh monoklonal'nymi antitelami.

The ultrastructure of different human blood T-lymphocyte subpopulations was studied by immunoelectron microscopy using "Orthoclone" monoclonal antibodies (OKT-3, OKT-4 and OKT-8) and peroxidase-antiperoxidase (PAP) complex. It was shown that T-lymphocyte subpopulations, distinct in their surface markers and function, possessed some specific features of submicroscopic structure. OKT-4+ cells (helper/inductor) were characterized by small size, high nucleus/cytoplasm ratio and few cytoplasmic organelles. OKT-8+ cells (suppressor/killer) were larger, had low nucleus/cytoplasm ratio and bean-like nucleus, with their cytoplasm containing numerous mitochondria, well-developed Golgi complex and osmiophilic granules.

[Comparative immunological and electron microscopic study of the structure of E-RFC and OKT-3+ lymphocytes from human blood]. / Sravnitel'noe immunoélektronno-mikroskopicheskoe issledovanie stroeniia E-ROK i OKT-3+ limfotsitov krovi cheloveka.

Methods of immune electrone microscopy have revealed that OKT-3+ cells and E-RFC of human blood have a similar ultrastructure. Definite morphologic spectrum is observed within each of these fractions, that testifies to the heterogeneity of surface markers of circulating T-lymphocytes and their ultrastructure.

[Electron microscopy and cytochemistry studies of lymphocyte subpopulations in corticosteroid deficiency]. / Elektronno-mikroskopicheskoe i tsitokhimicheskoe issledovanie subpopuliastsii limfotsitov krovi pri nedostatke kortikosteroidov.

The authors studied the number, structure and function of different blood T lymphocyte subpopulations in experimental hypocorticoidism of male CBA mice with the use of modern hematological, cytochemical, immunological and electron microscopy methods. One week after bilateral adrenalectomy the blood showed a considerable absolute and relative lymphocytosis determined by an increase in the content of Thy I-positive, ANAE-positive (giving positive reaction during identification of acid non-specific alpha-naphthylacetate esterase) cells and large granule-containing lymphocytes, a group, which includes natural killers and cells possessing antibody-dependent cell-mediated cytotoxicity. Transmission and scanning electron microscopy demonstrated changes in submicroscopic structure of different blood lymphocytes in adrenalectomized mice. It may be inferred that during hypocorticoidism, there takes place a complicated immunological transformation resulting in the changes in the function of different T lymphocyte subpopulations and natural killers responsible for immune defence of the body.

Immunoelectron microscopy of the bone marrow mononuclears labelling with rabbit anti-mouse brain serum using peroxidase-anti-peroxidase method.

Ultrastructural features of mouse bone marrow mononuclears (BMM) labelled with peroxidase-anti-peroxidase (PAP) complex using rabbit anti-mouse brain serum (RAMBS) and rabbit anti-mouse brain serum, absorbed with thymocytes (RAMBST) were investigated. After incubation of cells with RAMBST which have high activity against mouse pluripotent hemopoietic stem cells (CFUs) about 0.6% of BMM were labelled. According to their fine structure these cells were similar to those described earlier as presumptive pluripotent hematopoietic stem cells (PHSC). After incubation with RAMBS 1.9% of BMM were labelled, including cells marked by RAMBST and typical small lymphocytes with ultrastructure of T-lymphocytes.

[Ultrastructure, metabolism and function of the white blood cells in the process of the formation of an immune response and in exposure to glucocorticoids. II. The effect of immunization on the number and sub-microscopic structure of granulocytes in rabbits]. / Ul'trastruktura, metabolizm i funktsiia kletok beloi krovi v protsesse formirovaniia immunnogo otveta i pri vozdeistvii na nego gliukokortikoidov. Soobshchenie II. Vliianie immunizatsii na chislo i submikroskopicheskoe stroenie granulotsitov krovi krolikov.

The absolute count of different types of granulocytes was determined and their ultrastructure was studied in the blood of rabbits immunized with bovine serum albumin. Both the primary and repeated antigen administration were followed by regular quantitative and qualitative changes in neutrophils, basophils, and eosinophils whose degree and character were determined by the stage of the immune response formation.

[Ultrastructure, metabolism and performance of white blood cells during development of the immune response and the effect of glucocorticoids on it. I. Effect of immunization on the number and submicroscopic structure of lymphocytes and monocytes in the blood of rabbits]. / Ul'trastruktura, metabolizm i funktsiia kletok beloi krovi v protsesse formirovaniia immunnogo otveta i pri vozdeistvii na nego gliukokortikoidov. Soobshchenie I. Vliianie immunizatsii na chislo i submikroskopicheskoe stroenie limfotsitov i monotsitov krovi krolikov.

As a result of immunization and reimmunization with bovine serum albumin there was a change of the number and correlation of various morphologically identifyable species of lymphocytes in the blood of rabbits. Regular submicroscopic changes in the nucleus and organoids of the cytoplasm pointing to increased functional activity of these cells were revealed at the ultrastructural level in many circulating lymphocytes belonging to various populations.