[Adaptation of different modifications of enzyme immunoassay and immune blot assay to assess the candidate vaccine HIVREPOL against HIV/AIDS].

A preclinical trial of the vaccine HIVREPOL provided a complex of methods for assessing the identity and specific activity of vaccines against HIVIAIDS. The identity of "HIVREPOL" has been assessed by indirect enzyme immunoassay (EIA): the vaccine specifically binds the antibodies of the sera from HIV-infected individuals. Immune blot assay was the most informative method for assessing the identity of the candidate vaccine. The sera from HIVREPOL-vaccinated mice recognized the proteins gp41, p24, p55 of cultured HIV1 on "New-Lay-Blot1" strips. The bands corresponding to p24 were revealed in the line blots "Blot-HIV-1/2+O" and "INNO-LIA-HIV-Confirmation". The specific activity of the HIVREPOL vaccine was confirmed from the reactivity of sera of the mice vaccinated with recombinant proteins of the immunosorbents available in EIA test systems for the detection of HIV antibodies. Competitive EIA established the antigen-binding activity of sera from HIVREPOL-vaccinated mice against the native reference HIV-1 antigen.

[Dynamics of disorders of cellular and humoral immunity parameters in patients with insulin-dependent diabetes mellitus]. / Dinamika narushenii pokazatelei kletochnogo i gumoral'nogo immuniteta u bol'nykh insulinzavisimym sakharnym diabetom.

Analysis of immunocompetent cell subsets in peripheral blood of patients with insulin dependent diabetes mellitus (IDDM) and the determination of sICA-autoantibodies in their sera were performed by flow rate cytometry and compared to healthy donors and patients with noninsulin-dependent diabetes mellitus (NIDDM). It was shown that newly diagnosed IDDM was characterized by predominant disturbances of humoral immunity, and disease progression was mainly accompanied by cellular immunity disturbances. Exogenous insulin was one of the causes of such disturbances. A tendency to normalization of cellular rather than humoral immunity was observed after the onset of human monocompetent therapy of IDDM patients. It is likely that the appearance in the peripheral blood of activated T-lymphocytes accompanied by sICA-autoantibodies and increased mature B-lymphocytes and NK-cells counts corresponds to increased ADCC against pancreatic beta cells in IDDM development.

Pancreatic antigenic complex p64 69: involvement in regulation of insulin secretion and relation to glutamic acid decarboxylase.

The role of pancreatic beta-cell antigenic structures in modulation of insulin secretion in vitro was recently demonstrated by others. Here we report generation of a monoclonal antibody (mAB) ICA-1 to non-species specific beta-cell antigen(s) 64, 67 and 69 kDa. The mAB inhibits glucose stimulated insulin secretion in islet cell cultures. The ability of mAB ICA-1 to immunoprecipitate active glutamic acid decarboxylase from high speed supernatants of pancreatic and brain crude extracts was demonstrated. The 64, 67 and 69 kDa antigenic material was affinity purified from pancreatic islet cell high speed supernatants, active glutamic acid decarboxylase was found in the material. Immunoaffinity purification with mAB ICA-1 of GAD-like pancreatic beta-cell antigenic material has provided evidence of possible involvement of glutamic acid decarboxylase in modulation of insulin secretion.

Centriolar location during blood cell spreading and motion in vitro: an ultrastructural analysis.

Different motile blood cells behave differently on a glass surface. Macrophages go through all the stages of spreading described previously for fibroblasts; granulocytes become polarized after a short stage of radial spreading, while the polarization of lymphocytes takes place immediately upon attachment. The active centrioles of the cells we have studied orient their distal ends towards the upper cell surface. The centrioles in blood cells have a different location with respect to the nucleus and the leading edge of the cell: in macrophages the centrioles are situated mostly anterior to or at the side of the nucleus; in granulocytes they lie between the nuclear segments; and in lymphocytes they are positioned strictly posterior to the nucleus, in the uropod. In each case, however, the centrioles are localized in the central region of the cytoplasm. Their alignment does not appear to be related to the blood cells' random motion in vitro.

[Lines of transformed mouse thymus cells. II. Cell morphology, karyology, ultrastructure and growth in vitro and in vivo]. / Linii transformirovannykh kletok timusa myshi. II. Morfologiia, kariologiia, ul'trastruktura, rost kletok in vitro i in vivo.

Morphology, fine structure, karyology and growth of intrathymus pre-T-cell cultures (TC.SC-1/1.1 and TC.SC-1/2.0) were studied both in vitro and in vivo. The cultures were induced by injecting to mice a supernatant enriched with interleukin 2. The results obtained confirm the malignant transformation of cells of the lines obtained and the involvement of endogenic lymphotropic viruses in this process. The lines obtained are defective in hypoxanthine phosphoribosyltransferase. This property may serve as a basis for their use in hybridoma technology.

[Behavior of the cell center during epithelial cell spreading]. / Povedenie kletochnogo tsentra pri rasplastyvanii épitelial'nykh kletok.

In the epithelial cells of mouse embryo renal channels, centrioles are located near the plasma membrane of the apical part of the cell. In most of the cells an active centriole carries a cilium, which comes out into the channel lumen. In the epithelial cells, suspended after trypsinisation and in single cells adhering to the substrate, the centrioles are located near the nucleus, and the outcoming cilia are not observed. In the spread cells of epithelial islets, the centrioles are also found near the nucleus, and in most cases an active centriole carries a cilium, which comes out of the cytoplasm at the upper side of the cell. In the peripheral cells of the islet, centrioles are positioned between the nucleus and the active edge of the cell. In the epithelial cells in situ, a relatively small number of microtubules radiate from the active centrioles. In the suspended cells, the activation of microtubule formation is observed in the cell center. In the spread cells of the epithelial islets there occurs a further increase in the number of microtubules radiating from the active centrioles. In the peripheral cells which cause translocation of the epithelial islet in the culture, the number of microtubules, radiating from the centrioles does not differ significantly from that of the inner cells of the islet. The cell center of the epithelial cells does not seem to be actively involved in the locomotion of the epithelial cells in the culture.

[Cell center of macrophages, granulocytes and lymphocytes during in vitro cell spreading, polarization and movement]. / Kletochnyi tsentr makrofagov, granulotsitov i limfotsitov pri rasplastyvanii, poliarizatsii i dvizhenii keltok in vitro.

Different motile blood cells behave in a different way upon spreading on the glass surface. Macrophages pass through all the stages of spreading described for fibroblasts (Vasiliev, Gelfand, 1976); granulocytes are polarized after a short staying in badly spread conditions, lymphocytes are polarized immediately after setting of the glass surface. In relation to the leading edge and the cell nucleus, centrioles in the described cell types are located differently. In macrophages they are mainly in the front or on one side of the nucleus, in granulocytes they lie within the ring-like nucleus, in lymphocytes they are strictly located behind the nucleus in the uropode. In all the cases, however, centrioles are localized in the central region of the cytoplasm. Their location does not appear to be connected with the movement direction of blood cells. The distal ends of the active centrioles are faced to the upper cell surface in the examined cells. It is suggested that the centriole can distinguish the free cell surface and the surface associated with the substrate.

[Structural change in the cell center during fibroblast polarization and movement]. / Izmenenie struktury kletochnogo tsentra pri poliarizatsii i dvizhenii fibroblastov.

In polarizing and migrating 3T3/Balb mouse fibroblasts, the centrioles are located between the nucleus and the leading edge of the cell. In cells within the monolayer and in migrating cells, the centrioles have a random orientation towards the substrate. In polarized cells, that still remain in the monolayer, one centriole may be perpendicular to the substrate plane in 70% of cases. Upon polarization and migration of fibroblasts, the number of microtubules, which radiate from the centriolar region, increases. These data support a hypothesis that the number of microtubules in the cell centre characterizes the rate of their renovation in the cytoplasm. It is concluded that the cell centre is strongly involved in polarization and migration of fibroblasts.

[Behavior of the cell center in the distribution of fibroblasts]. / Povedenie kletochnogo tsentra pri rasplastyvanii fibroblastov.

In suspension state of murine embryo fibroblasts the long microtubules radiating from the cell centre disappear, but many short microtubules are found around the centrioles. The growth of long microtubules begins after the cell attachment to the substrate. The number of long microtubules increases and reaches the maximum in polarized fibroblasts. At the radial spreading stage the number of microtubules increases as moving off the centriole. This fact disagrees with the statement that all microtubules in the cell are connected with the cell centre, but agrees with the hypothesis of the conveyor microtubule assembly. After the cell attachment to the substrate the centrioles move to the part of the cell facing the substrate. At the radial spreading stage the active centrioles are settled down mainly perpendicular to the plane of the substrate. Such orientation is kept after the treatment of the radially spread cells with cytochalasin B and disappears after disruption of microtubules by colcemid. It is supposed that the orientation of the active centrioles perpendicular to the substrate is connected with the formation of long microtubules during the process of spreading of fibroblasts.