concanavalin-a and Leukemia--Erythroblastic--Acute

Ganglioside expression on cytotoxic T lymphocytes induced against the mouse erythroleukaemia line FBL-3N, was analysed, compared with that of naive T lymphocytes in the thymus, lymph nodes and spleen. Although normal uncultured and cultured T lymphocytes expressed no GD2, GD3 or GM2 gangliosides, cytotoxic T cells with CD4+CD8-, CD4-CD8+, and CD4-CD8- phenotypes reacted with anti-GD2 monoclonal antibody with various intensities. The reactivity with anti-GD2 was associated with the intensity of cytotoxic activity as analysed using cytotoxic T lymphocyte (CTL) clones established from the bulk CTLs of each phenotype. An increase of the mRNA expression of GM2/GD2 synthase gene was demonstrated by Northern blot hybridization using RNA extracted from thymocytes, spleen cells, Con A-treated spleen cells and various types of CTL cells. These results indicated that GD2 ganglioside expression might be associated with the functional differentiation of murine T lymphocytes.

Topics: Animals; Antibodies, Monoclonal; Blotting, Northern; Concanavalin A; Cytotoxicity Tests, Immunologic; Female; Flow Cytometry; G(M2) Ganglioside; Gangliosides; Leukemia, Erythroblastic, Acute; Male; Mice; Mice, Inbred C57BL; N-Acetylgalactosaminyltransferases; Polypeptide N-acetylgalactosaminyltransferase; T-Lymphocytes, Cytotoxic; Tumor Cells, Cultured; Up-Regulation

Certain tumor cells generate factors that inhibit neutrophil chemotaxis. Our study was designed to explore whether such factors are produced by K 562 malignant cells and whether these have a broader effect in altering neutrophil functions. After 48 h of in vitro culture of K 562 cells, the culture medium and the cells were separated, lyophilized, and extracted with ethanol. These K 562 products, i.e., either the cell or supernatant extract, inhibited both nonstimulated locomotion and locomotion induced either by FMLP or activated serum. Furthermore, K 562 products inhibited neutrophil adherence and oxidative burst induced by opsonized zymosan, whereas oxidative burst induced by PMA or FMLP was not altered. K 562 products had an inhibitory effect on the PMN binding to iC3b-coated particles. They did not modify Mo1 expression of resting cells, did not alter the up-regulation of the receptor induced by FMLP but inhibited the FMLP-induced capping of Mo1 Ag. Con A capping was also inhibited. Actin polymerization in FMLP-stimulated PMN, as measured by flow cytometry and phalloidin binding to F-actin, was inhibited by K 562 products. The inhibitory factor present in K 562 products (cell and culture supernatant) was purified in three steps including gel filtration, ion-exchange chromatography, and IEF. The eluted active fraction corresponded to single band of about 8 kDa on SDS-PAGE. From these experiments, it is concluded that K 562 malignant cells in culture contain and release a low molecular mass factor (congruent to 8 kDa) that inhibits all adherence-related functions of neutrophils, whereas it does not alter FMLP- or PMA-induced oxidative burst. Further studies are needed to assess whether products of other tumor cells also act on the neutrophil by inhibiting adherence-related functions, Mo1 function and capping, and actin polymerization.

Topics: Actins; Cell Adhesion; Cell Movement; Chemotaxis, Leukocyte; Concanavalin A; Humans; Leukemia, Erythroblastic, Acute; Molecular Weight; Neutrophils; Oxygen Consumption; Peptide Hydrolases; Receptors, Complement; Receptors, Concanavalin A; Superoxides; Tumor Cells, Cultured

The effect of TBG on the functional activity of different cell lines, spontaneous and Con A induced proliferation of PBL was studied. If concentration of TBG is higher than 50 mu kg/ml it suppresses the proliferation in many used cell lines, except choriocarcinoma and cancer of uterus. The reliable increasing of spontaneous proliferation of PBL, Jurkat and K-562 cells may be observed if concentration is more lower (0.5-15 mu kg/ml). However proliferation of other cell lines corresponds to control level, and Con A induced proliferation of PBL is inhibited. The effect was more marked at 48, as compared to 24 hours of cell incubation with TBG.

Topics: Biomarkers, Tumor; Cell Division; Cell Line; Choriocarcinoma; Concanavalin A; Female; HeLa Cells; Humans; Leukemia, Erythroblastic, Acute; Leukemia, Myeloid; Leukemia, T-Cell; Lymphocytes; Lymphoma, B-Cell; Lymphoma, T-Cell; Pregnancy-Specific beta 1-Glycoproteins; Tumor Cells, Cultured; Uterine Neoplasms

In this paper we present first the results of our most recent investigations on gravitational effects on the activation of human lymphocytes: by immunoenzymatic staining and by using concanavalin A (Con A) coated to red blood cells (RBC) we demonstrate that the increase of activation measured at 10xg is due to a simultaneous activation of T- and B-lymphocytes whereas at 1xg only T-cells are stimulated. Conversely, activation of T-cells by chemical modification of the membrane with sodium periodate is depressed at 10xg. Secondly, experiments performed in the centrifuge as well as in the clinostat with Friend, K-562, and hybridoma cells show that each cell line develops its own adaptation reaction to gravitational stress.

Topics: B-Lymphocytes; Cells, Cultured; Centrifugation; Concanavalin A; Dimethyl Sulfoxide; Erythrocytes; Friend murine leukemia virus; Hemin; Humans; Hybridomas; Hypergravity; Leukemia, Erythroblastic, Acute; Lymphocyte Activation; Mitogens; Periodic Acid; Rotation; T-Lymphocytes; Tumor Cells, Cultured

This paper reports on the characteristics of killing by a human and a murine tuberculin (PPD)-specific T helper clone of targets to which PPD was attached via the lectin concanavalin A (Con A). The killing was specific for PPD from M. tuberculosis; and targets coupled to Con A alone or to PPD from M. paratuberculosis were not killed. Target cells carrying Con A-PPD were more effectively lysed than PPD-pulsed cells. This form of lymphocyte killing, though highly significant, was inefficient. Maximum killing of PPD carrying targets was 30-40% at effector to target ratios of 20:1 and at 16 h. Cells carrying 2 x 10(6) molecules of PPD and less than 1.5 x 10(6) molecules Con A per cell were killed most efficiently. A major distinction between this helper T cell killing and that mediated by cytotoxic T cells was that both TH clones displayed bystander lysis and killed PPD uncoupled targets when these were cultured with syngeneic PPD-bound targets. This suggests that the mechanism of cytotoxicity may involve soluble mediators.

Topics: Animals; Cell Line, Transformed; Clone Cells; Concanavalin A; Cytotoxicity, Immunologic; Dose-Response Relationship, Immunologic; Epitopes; Fibrosarcoma; Herpesvirus 4, Human; Humans; Leukemia, Erythroblastic, Acute; Mice; T-Lymphocytes, Helper-Inducer; Time Factors; Tuberculin; Tumor Cells, Cultured

Topics: Abrin; Animals; Cell Line; Concanavalin A; Drug Interactions; Friend murine leukemia virus; HeLa Cells; Humans; Leukemia, Erythroblastic, Acute; Leukemia, Experimental; Mice; Plant Proteins; Ricin

Friend erythroleukaemic cells can be induced to mature along the erythroid differentiation pathway when an inducing agent such as dimethyl sulphoxide is included in the medium. In the absence of the inducing agent, the 707B line of Friend erythroleukaemic cells is highly agglutinable by the lectins concanavalin A or wheat germ agglutinin. However, 48 h after the induction of differentiation, there is a marked decrease in the agglutination of the cells in the presence of either lectin. This suggests that early in differentiation a change occurs in the cell membrane preceding the onset of globin synthesis which starts approximately 72 h after induction. The change in agglutination by concanavalin A also occurs in the presence of reagents which do not induce haemoglobin synthesis in the 707B line of Friend erythroleukaemic cells but which are able to stimulate the synthesis of this protein in other erythroleukaemic cell lines. The reduction in the agglutinability of the differentiating cells does not seem to result from a reduction in the number of concanavalin A receptors on the cells, nor does it reflect a change in the clustered distribution of concanavalin A receptors in the differentiating cells. Both the control and dimethyl sulphoxide-induced cells show a similar patchy distribution of ferritin-labelled concanavalin A when examined by electron microscopy. Polyacrylamide gel electrophoresis shows little change in the total pattern of protein synthesis by control and differentiating cells when pulse-labelled with [35S] methionine. However, use of 125I-labelled concanavalin A to stain polyacrylamide gels, on which the total proteins of control and differentiating cells had been separated, revealed a profound change in the composition of the concanavalin A-binding proteins. The control, undifferentiated cells contained eleven or more classes of concanavalin A-binding glycoproteins, many of which stained to a lesser degree as the cell density increased. After the onset of differentiation, 2 new concanavalin A-binding glycoproteins appeared within 48 h. One of these proteins has a molecular weight in excess of 180 000 while the other migrated with an apparent molecular weight of approximately 100 000. After erythroid differentiation had progressed for 120 h, these newly synthesized glycoproteins became the major concanavalin A-binding proteins of the erythroleukaemic cells.

Topics: Cell Aggregation; Cell Differentiation; Cell Line; Cell Membrane; Concanavalin A; Dimethyl Sulfoxide; Electrophoresis, Polyacrylamide Gel; Erythrocytes; Lectins; Leukemia, Erythroblastic, Acute; Microscopy, Electron; Receptors, Concanavalin A

The sensitivity to agglutination by several plant lectins has been studied during the induced erythroid differentiation of Friend erythroleukemic cells in culture. In addition, the number of lectin receptors on the cell has been measured. It is shown that early during the differentiation, there is an increase in agglutinability while the receptor density remains constant. In the later phase of the differentiation process, the cells lose their sensitivity to agglutination while the receptor number and density increases. These changes were not observed on nonerythroid mastocytoma culture cells. Two nondifferentiating variants of the FL cells were shown to have altered sensitivities to agglutination by ConA.

Topics: Agglutination; Binding Sites, Antibody; Cell Differentiation; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Dimethyl Sulfoxide; Friend murine leukemia virus; Lectins; Leukemia, Erythroblastic, Acute; Receptors, Concanavalin A