[Heterogeneity of cell population of lymphoma NK/Ly and leukemia L-1210 according to carbohydrate structure of cell surface: immunocytochemical analysis of lectin binding].

The heterogeneity of tumor cell populations according to binding of lectins from lentil (LcL), wheat germs (WGA), peanut (PNA) and concanavalin A was investigated on a model of murine Nemeth-Kellner lymphoma (NK/Ly) and leukemia L-1210. Bound lectins were detected by indirect immunochemical method using home obtained polyclonal antilectin antibodies and immunogold silver staining (IGSS) technique. Significant differences in binding of Con A were revealed between NK/Ly (67% Con A+) and L-1210 (7.2% Con A+) cells, while the differences in binding of other lectins with both types of tumor cells were not significant. A relatively high percentage of PNA+ cells was registered that can indicate a high degree of desialization of membrane glycoproteins.

In vitro studies of activation of phagocytic cells by bioactive peptides.

The effect of formyl chemotactic peptide (fCTP, fMet-Leu-Phe), beta-amyloid peptides (beta-AP, 1-42, 1-16 and 25-35), and bradykinin (BK) on functional activity of phagocytic cells has been investigated. Wheat germ agglutinin (WGA) was also used as a model membrane binding agent of polypeptide nature. Murine monocyte-macrophage cell line J774.2 and normal human blood polymorphonuclear (PMN) cells were used as target phagocytic cells. Their activity was quantitatively estimated by measuring phagocytosis of killed yeast cells. Beta-AP (1-41) maximally stimulated phagocytosis at 0.1 microg/ml, BK--at 1.0 microg/ml, and fCTP--at 2.0 microg/ml. Beta-AP (1-16) and beta-AP (25-35) were inactive in used test-systems. Phagocytosis-inducing activity of beta-AP (1-42) and BK reached maximal levels in 2 h and decreased after 4-6 h of incubation. Phagocytosis numbers were compared with the indicators of phagocytic cell activation, such as absorption of neutral red dye, glucose utilization, production of super-oxide anion (NBT-test) and nitrite accumulation (indicator of NO production). NBT-test, which may be related to the killing ability of phagocytic cells towards the ingested objects, was positive only in stimulated PMN leukocytes, while the nitrite accumulation was detected only in stimulated macrophages. Nitrite accumulation in macrophages was markedly induced by lipopolysaccharide and to a lower extent by 0.5 microg/ml beta-AP (1-42). In high dose (5.0 microg/ml) beta-AP suppressed nitrite accumulation in macrophages stimulated by lipopolysaccharide. Other studied peptides were inactive in inducing nitrite accumulation. Transforming growth factor type beta suppressed phagocytic activity of PMN cells activated by beta-AP or WGA. The anti-inflammatory drugs (indomethacin and L-lysine aescinate) inhibited beta-AP (1-42)-induced phagocytosis. The interrelations between the regulatory pathways of BK, beta-AP and fCTP are discussed.

[The preparation of an anti-A reagent from the lectin of hairy vetch (Vicia villosa) for detecting antigen A in human erythrocytes and saliva]. / Izgotovlenie reagenta anti-A iz lektina goroshka mokhnatogo dlia vyiavleniia antigena A v éritrotsitakh i sliune cheloveka.

Anti-A reagent for the detection of the relevant antigen in human red cells and saliva has been developed. The reagent is prepared by dissolution of purified Vicia villosa lectin in the serum of group AB or B diluted 8 times with PBS. This solvent appreciably improves the anti-A selectivity of lectin and is used for titration. Purification of lectin by ammonium sulfate precipitation is described. The reagent agglutinates group A red cells in the minimal lectin concentration of 2.5 to 10 micrograms/ml and does not agglutinate group 0 and B red cells in concentration 10 mg/ml. Moreover, the reagent is fit for the detection of antigen A in traces of blood and saliva by the absorption-elution method.

Electron cytochemical study of carbohydrate components in different types of cultured glial cells of snail Helix pomatia.

Using a variety of colloidal gold-labelled lectins with different sugar specificities, the structure and topography of carbohydrate determinants of the surface membrane of in vitro cultured glial and nerve cells of the snail Helix pomatia have been electron cytochemically studied. Heterogeneity of carbohydrate pools among different types of glial cells and between glial and nerve cells was established. It was found that satellite glial cells having the ultrastructural signs of cells with high metabolic level (type II cells) selectively bind GNA which is specific to terminal alpha-D-mannose residues and do not bind other mannose-specific lectins, Con A and LCA. GNA determinants are absent in satellite type I glial cells, fibrous glial cells, microglia and neurons. It has been found that glial cells (satellite type I and II glial cells, filamentous glial cells and microglial cells) do not bind PVA and LABA. LTA did not bind to any glial cells and binds weakly to neurons. Con A and WGA determinants which are abundant on the neurons are completely absent on satellite type II glial cells but present on satellite type I glial cells and filamentous glial cells. Microglial cells contain Con A and LCA determinants and the density of PNA determinants on these cells is the highest compared to other types of glial cells or neurons. It is concluded that some lectin determinants (for RCA-1, PNA, LPA) are present on all types of glial cells, while another determinant (GNA) is specific for a definite type of glial cells and can serve as a marker of these cells. The role of specific carbohydrate determinants in the functioning of a neuron-glial complex is discussed.

[Electron-cytochemical study of carbohydrate components in the surface membrane of cultured Helix pomatia neurons]. / Elektronno-tsitokhimicheskoe issledovanie karbogidratnykh komponentov poverkhnostnoi membrany kul'tiviruemykh neironov vinogradnoi ulitki.

The structure and topography of carbohydrates on the surface of nerve cells of snail Helix pomatia cultured in vitro have been characterized with a series of colloidal gold-labelled lectins of different sugar specificity. The analysis of the lectin binding has shown substantial differences in the carbohydrate pattern between the soma of monoaminergic and peptidergic neurons. It has been found that the surface of monoaminergic and peptidergic neurons contains N-acetylglucosamine (WGA+) and N-acetyllactosamine (RCA-1-) determinants and does not contain neuraminic acid (LPA-) and complex branched N-glycosyl chains (PVA-). At the same time N-acetylgalactosamine (HPA+) was detected on the peptidergic neuron membrane only. It has been concluded that terminal residues of sialic acid are absent on the most of snail nerve cells. Differences in lectin binding between monoaminergic and peptidergic neurons can serve as a basis for formation of specific connections of cells by different types in the developing brain.

[The characteristics of lectin binding to the surface of differentiated neuroblastoma C 1300 cells]. / Osobennosti sviazyvaniia nekotorykh lektinov s poverkhnost'iu differentsirovannykh kletok neiroblastomy C 1300.

Peculiarities of the binding of lectins of castor bean, cochlea, soya bean, lime bean and wheat germs to the surface of differential neuroblastoma C 1300 N 18 cells have been studied using the method electron cytochemistry. It is found that the quantity of the bound lectin conjugates with colloidal gold on the surface of differentiated cells varied considerably from that on the surface of nondifferentiated cells.

[Determination of lectin receptors on the surface membranes of lymphoid cells]. / Opredelenie retseptorov lektinov na poverkhnostnykh membranakh limfoidnykh kletok.

The authors describe a method for the detection of carbohydrate determinants contained by the cellular surface membrane glycosylated biopolymers, based on the use of horse radish peroxidase-conjugated Arachis, Helix pomatia, Ricinus communis, Lens, Glycine max, Sophora japonica lectins. The pattern of lectin receptor distribution on normal human blood lymphocytes, tonsillar lymphoid cells, and in some forms of leukemia and lymphoma is shown. The authors consider it essential that a lectin kit be used along with enzymochemical and immunocytochemical markers as an additional test for the diagnosis of various forms and types of malignant lymphoproliferative diseases.

[The relation of changes in the surface markers and the induction of thymocyte functional activity under the action of humoral thymic factors]. / Vzaimosviaz' izmenenii poverkhnostnykh markerov i induktsii funktsional'noi aktivnosti timotsitov pri deistvii gumoral'nykh faktorov timusa.

Mouse thymocytes were fractionated on PNA+- and PNA-cells. The PNA+-fraction was treated by tactivin--a preparation of thymic hormones. The treatment resulted in the loss of PNA-receptors from the surface of 10-12% of PNA+-cells. The treated suspension was again fractionated by the same method. PNA-thymocytes derived from both the first and the second procedure of fractionation expressed less Thy-1.2 antigen and SBA-receptors than PNA+-cells. Primary and induced PNA-thymocytes were able to produce IL-2 after mitogen stimulation. The data can be interpreted as an evidence of the ability of thymic hormones to induce not only a phenotypical but also a functional maturation of PNA+-thymocytes and of linkage of all the observed signs of maturation.

[Study of the carbohydrate components of the surface of S 1300 neuroblastoma cells with the use of cytochemical and lectin agglutination methods]. / Issledovanie uglevodnykh komponentov poverkhnosti kletok neiroblastomS 300 s pomoshch'iu metodov tsitokhimii i agliutinatsii lektinami.

The carbohydrate determinants of the cell surface of neuroblastoma C 1300 clone N 18 have been investigated by light microscopy with a panel of 11 lectins as cytochemical reagents. The expression of N- and O-glycanes with a predominance of 3-4 antigenic N-glycosyl chains was revealed. The characteristic feature of neuroblastoma cells is an abundance of an antigen A surface determinant. A strong correlation is shown between the intensity of lectin binding and index of cell agglutination. After the binding of lectins with living cells the lectin-receptor complexes are clustered, endocytosed and for 4 hours are concentrated as microvesicles in the cytoplasm compact area. Heterogeneity of the neuroblastoma cell population on the binding of peanut and Lens culinaris lectins is revealed.

[Identification and properties of insulin receptors in neuroblastoma C 1300 N 18 cells in various functional states]. / Identifikatsiia i svoistva insulinovykh retseptorov v kletkakh neiroblastomy C 1300 N 18 pri razlichnykh funktsional'nykh sostoianiiakh.

The presence of insulin receptors in neuroblastoma C 1300 N18 cells was shown by the method of the [125I]insulin binding with cells as well as by the electron-cytochemical methods based on the analysis of the binding of colloid gold-labelled insulin and of agglutinin of wheat embryos with the surface of plasma membrane. It is established that the number and distribution of insulin receptors depend on the functional state of cells. Expression of receptors on the membrane increases after 5'-deoxyuridine-induced differentiation of cells. Due to changes in the lipid composition of cells caused by the incubation with lecithin-cholesterol liposomes (an increase in the content of cholesterol and its esters, as well as of unsaturated fatty acids and appearance of lysolecithin) the quantity of insulin receptors decreases and cell membranes are damaged. The lowering insulin regulation is not observed for insulin receptors of the neuroblastoma C 1300 N18 cells.

[Lectin receptors--histochemical markers in the study of leukemias and lymphomas]. / Retseptory lektinov--gistokhimicheskie markery v izuchenii leikozov i limfom.

Data are presented on histochemical determination of lectin receptors on surface membranes of different haematopoietic and lymphoid cells of healthy persons and of patients with leukemias and lymphomas.

[Study of membrane glycoproteins of human erythrocytes using lectins]. / Issledovanie membrannykh glikoproteinov éritrotsitov cheloveka s primeneniem lektinov.

A method to study the glycoprotein composition of cell membranes, in particular of human red blood cells, has been developed. It includes the separation of membrane components by the SDS-polyacrylamide gradient slab gel electrophoresis, electroblotting of the phoretograms onto the nitrocellulose sheets and detection of glycoprotein fractions with FITC and peroxidase labeled lectins. PNA detected asialoglycoproteins with O-linked oligosaccharide chains, corresponding to all the PAS-positive bands of the phoretogram. SBA interacted more selectively and revealed only certain PAS-positive bands. Glycoproteins with N-linked carbohydrate chains were PAS-negative and can be identified only by the interaction with WGA, LCL, RCA. Group-specific agglutinins have shown that the ABO antigenic determinants are located in N-linked carbohydrate chains of membrane glycoproteins.

[Evaluation of the distribution of lectin receptors on the surface of tumor and transformed cells using methods of variation statistics]. / Otsenka raspredeleniia lektinovykh retseptorov na poverkhnosti opukholevykh i transformirovannykh kletok s pomoshch'iu metodov variatsionnoi statistiki.

Statistical procedures were used to estimate lectin receptor distribution on the surface of ascite lymphoma cells, neuroblastoma C-1300 cells and of transformed human T- and B-derived lymphoid cell lines. Relationships between the arithmetic means and mean square variances for sample populations from each cell and ferritin- or colloidal gold-lectin combination were used to define four types of topographical distributions: uniform-ordered, uniform-random, random and clustered.

[Lectin receptors in the salivary glands of the rat during postnatal ontogenesis]. / Restseptory lektinov v sliunnykh zhelezakh krys v protsesse postnatal'nogo razvitiia.

Distribution of lectin-binding sites in rat submandibular and sublingual salivary glands during postnatal development has been investigated. Lectin preparations include con A, lentin lectin, castor beans agglutinin, peanut, soybean and Sophora japonica agglutinins, wheat germ agglutinin and lectin from the bark of Laburnum anagyroides. The direct and indirect peroxidase techniques are used. According to the similarities of histochemical patterns, all lectins are divided into four groups. Besides the general patterns of lectin binding sites, some details are noted. Lectins of peanut and Sophora japonica possess an extremely high affinity to mast cells, con A, lens lectin, castor beans and wheat germ agglutinins--to serous demilunes cells. Laburnum lectin--to salivary ducts epithelia in adult rat salivary glands. Lentin lectin, con A and Laburnum lectin preferentially stain cells with specific granularity in granular ducts at early stages of postnatal development. Considering the character of staining, we propose for further histochemical investigations of the salivary glands lentin lectin, peanut agglutinin, wheat germ agglutinin and Laburum anagyroides lectin.

[Localization of lectin receptors on the surface of C1300 neuroblastoma cells]. / O lokalizatsii retseptorov lektinov na poverkhnosti kletok neiroblastomy C 1300.

D-mannose, D-galactose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine and L-fucose which are sugar determinants of receptors were found on the surface of neuroblastoma cells by means of four carbohydrate-specific lectin groups. Labeling of lectins was performed by horseradish peroxidase, ferritin and colloidal gold. Peculiarities of the lectin receptors distribution on the surface of immature neuroblastoma cells were detected.

[Cell surface receptors in lymphoid lines studied using lectins]. / Issledovanie retseptorov na poverkhnosti kletok limfoidnykh linii s pomoshch'iu lektinov.

The nature and distribution of some carbohydrate receptors on the cell membrane of transformed human B- and T-derived lymphocyte cell lines (RPMI-1788, Daudi, Jurkat, Molt-4) are studied by Ricinus communis agglutinin, soybean agglutinin, Helix pomatia lectin, peanut agglutinin and wheat germ agglutinin conjugated with ferritin and colloidal gold. The quality of receptors and the manner of their distribution detected by different lectins are considerably different.