concanavalin-a and Carcinoma--Ehrlich-Tumor

Conditional growth inhibition of murine Ehrlich carcinoma (EC) and B16 melanoma (B16Mel) was observed, following treatment of mice (Swiss and C57BL/6) with aqueous extract of neem (Azadirachta indica) (1 unit/mice/week for 4 weeks) either before or after inoculation of 1 x 10(6) tumor cells. Tumor inoculation after weekly injections for 4 weeks with neem leaf preparation (NLP) induced significant reduction of tumor growth (both EC and B16Mel) and increased survivability of mice. On the other hand, NLP treatment after tumor inoculation demonstrated no tumor growth inhibition in the NLP treated group in comparison to the PBS treated control. No direct cytotoxic effect of NLP towards EC and B16Mel tumor cells was observed in vitro. The spleen cells of NLP treated mice when mixed with inoculum of B16Mel tumor cells and injected into a group of mice, tumor growth was found to be significantly reduced and survivability of the tumor hosts increased remarkably in comparison to mice inoculated with tumor along with normal spleen cells. Concanavalin A (ConA) induced proliferation of lymphocytes from NLP treated mice was significantly higher than the lymphocytes of untreated mice. In in vitro, NLP by itself had no proliferative effects on lymphocytes but it co-stimulated ConA induced mitogenesis. NLP induced lymphocytosis as evidenced by increased lymphocyte count in blood as well as spleen. Flow cytometric evidence suggested that increase in CD4+ and CD8+ T cells accounted for lymphocytosis. The conditional tumor growth retardation, observed in mice treated with NLP before tumor inoculation, may be regulated by NLP mediated immune activation, having prominent role in the cellular immune function of the tumor host.

Topics: Animals; Anticarcinogenic Agents; Azadirachta; Carcinoma, Ehrlich Tumor; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Cell Proliferation; Concanavalin A; Female; Lymphocytes; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Phytotherapy; Plant Extracts; Plant Leaves; Spleen

The present study examined the effects of MAPA, an antitumor aggregated polymer of protein magnesium ammonium phospholinoleate-palmitoleate anhydride, isolated from Aspergillus oryzae, on concanavalin A (Con A)-induced spleen cell proliferation, cytokine production and on natural killer (NK) cell activity in Ehrlich ascites tumor-bearing mice. The Ehrlich ascites tumor (EAT) growth led to diminished mitogen-induced expansion of spleen cell populations and total NK activity. This was accompanied by striking spleen enlargement, with a marked increase in total cell counts. Moreover, a substantial enhancement in IL-10 levels, paralleled by a significant decrease in IL-2 was observed, while production of IL-4 and interferon-gamma (IFN-gamma) was not altered. Treatment of mice with 5 mg/kg MAPA for 7 days promoted spleen cell proliferation, IL-2 production and NK cell activity regardless of tumor outgrowth. In addition, MAPA treatment markedly enhanced IFN-gamma levels and reduced IL-10 production relative to EAT mice. A 35% reduction in splenomegaly with normal number of nucleated cells was also found. Altogether, our results suggest that MAPA directly and/or indirectly modulates immune cell activity, and probably disengages tumor-induced suppression of these responses. Clearly, MAPA has an impact and may delay tumor outgrowth through immunotherapeutic mechanisms.

Topics: Animals; Antineoplastic Agents; Aspergillus oryzae; Carcinoma, Ehrlich Tumor; Cell Proliferation; Cells, Cultured; Concanavalin A; Cytokines; Killer Cells, Natural; Linoleic Acids; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Mitogens; Organophosphorus Compounds; Spleen; Splenomegaly; Time Factors; Tumor Escape

Cytogenin recovered the reduced mitogenic response to Con A of spleen cells of tumor bearing mice in vitro. The suppressive factor(s) was detected in adherent cell population in spleen cells. The reduced antitumor effector activity of spleen cells taken from tumor bearing mice was also augmented by the treatment with cytogenin in vitro. The effect of cytogenin was neutralized by the treatment with anti-Mac 1 serum. Administration of cytogenin inhibited the production of nitric oxide by macrophages which is known as one of suppressor factors. Results indicate that one possible action of cytogenin exhibiting antitumor activity in tumor bearing mice may be due to modulation of Mac 1 positive cells.

Topics: Animals; Antibiotics, Antineoplastic; Carcinoma, Ehrlich Tumor; Cell Division; Concanavalin A; Coumarins; Female; Isocoumarins; Macrophages; Mice; Mice, Inbred ICR; Neoplasm Transplantation; Neoplasms, Experimental; Nitric Oxide; Spleen

The activity of gamma-glutamyl hydrolase, the enzyme which deglutamylates folyl and antifolyl polyglutamates, changed significantly in mouse cells during different phases of growth, being about two times lower in actively proliferating mice splenocytes and fibroblasts than in nondividing cells. In EAC cells growing in vivo the lowest activity was observed in cells in the logarythmic phase. Methotrexate treatment of mice in a dose of 500 mg/kg body weight increased the activity of the enzyme in EAC cells about 1.5 times. We suggest that gamma-glutamyl hydrolase is a proliferating dependent enzyme which together with folypolyglutamate synthetase ensures in cells an appropriate amount of folates in the form of polyglutamates necessary for optimizing folate-dependent biosynthetic activities.

Topics: 3T3 Cells; Animals; Carcinoma, Ehrlich Tumor; Cell Division; Cells, Cultured; Concanavalin A; Female; Fibroblasts; gamma-Glutamyl Hydrolase; Lymphocytes; Methotrexate; Mice; Spleen

Spleen cells from C57BL/6J mice bearing Ehrlich carcinoma growing as a solid tumor show progressive unresponsiveness to concanavalin A (Con A) and lipopolysaccharide (LPS) mitogens. This is accompanied by striking spleen enlargement with marked hematopoietic activity. Lymphoproliferative assays of normal spleen cells in co-culture with tumor-bearing spleen cells (TBSC) show that: (a) TBSC contain non-specific suppressor cells able to abrogate both Con A and LPS responses, or mixed lymphocyte reaction, of normal spleen cells and (b) suppression by TBSC is MHC-unrestricted, non-prostaglandin-mediated and greatly enhanced by Con A supernatants. Suppressor cells associated with TBSC are large, low-density cells without markers of mature B or T lymphocytes or of the mononuclear phagocyte system. Most appear to be asialo-GM1-negative, as suppression was only partially inhibited by treatment with anti-asialo-GM1 and complement. Since NK activity is lacking in TBSC, our data strongly suggest that these "null" suppressor cells are related to the natural suppressor (NS) cells found described in normal bone-marrow and neonatal spleens, or induced in adult spleens by total lymphoid irradiation, graft-vs.-host disease, or cyclophosphamide treatment.

Topics: Animals; Carcinoma, Ehrlich Tumor; Concanavalin A; G(M1) Ganglioside; Glycosphingolipids; Lymphocyte Activation; Male; Mice; Mice, Inbred Strains; Prostaglandins; Spleen; T-Lymphocytes, Regulatory

We investigated the in vitro binding of 125I-lectins to Ehrlich ascites tumor (EAT) cells and in vivo uptake of 125I-lectins in Ehrlich solid tumor (EST) bearing mice. In in vitro binding assays, phaseolus vulgaris agglutinin (PHA), pisum sativum agglutinin(PSA), and concanavalia agglutinin(Con A) showed a high affinity for EAT cells. The in vivo biodistribution of 125I-lectins showed 125I-I-PSA to be significantly taken up into EST tissues 24 h postinjection. After IV injection of 125I-PSA, uptake of the radioactivity into the tumor tissues reached a maximum at 6 h, and thereafter decreased. Rapid clearance of the radioactivity from blood and its excretion into kidney soon after injection of 125I-PSA were observed. When compared with the biodistribution of 67Ga-citrate in EST bearing mice 24 h postinjection, tumor to liver (T/B), tumor to muscle (T/M), and tumor to blood (T/B) ratios were superior for 125I-PSA. At 6 h postinjection, the T/B-ratio of 125I-PSA was 2.5, and this value may be sufficient to enable discernible diagnostic images. Our results suggest that PSA might be a useful tumor imaging radiopharmaceutical.

Topics: Animals; Binding Sites; Carcinoma, Ehrlich Tumor; Concanavalin A; Gallium Radioisotopes; Iodine Radioisotopes; Lectins; Male; Mice; Phytohemagglutinins; Plant Lectins; Radionuclide Imaging; Tissue Distribution

ConA-induced agglutination of Ehrlich ascites carcinoma (EAC) cells during development of the tumor in mice has been studied in vitro. EAC cells were incubated with different concentrations of ConA at 37 degrees C for various intervals of time and the cell aggregates of different sizes were scored under microscope. The older EAC cells (20-day-old) showed a decrease in cytoagglutination with respect to large aggregates formation compared to the younger (10-day-old) ones. The small aggregates formed by the 5-day-old EAC cells are dissociated by competition with alpha-methyl-D-mannoside but the large aggregates are not affected by the sugar treatment. The observed differences in the agglutination patterns are discussed in view of the influence of the lipid content of EAC cells and the ascites fluid on ConA-induced cytoagglutination.

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Aggregation; Concanavalin A; Lipid Metabolism; Methylmannosides; Mice

To investigate the involvement of actin filaments in concanavalin A (Con A)-induced cap formation and cytochalasin B (CB)-induced zeiotic knob migration, the distribution of F-actin was studied in Con A-treated and CB-treated Ehrlich ascites tumor cells (EATC) by fluorescence microscopy using heavy meromyosin conjugated with a fluorescent dye, N-(7-dimethylamino-4-methylcoumarinyl) maleimide, (DACM-HMM). In non-treated cells, the diffuse fluorescence of DACM-HMM was observed in the cytoplasm, particularly intensely under the plasma membrane and around the nucleus. In Con A- and CB-treated cells, the fluorescence was seen at Con A-induced-capped and CB-induced-knob-accumulated regions. This fluorescence was more intense in CB-treated cells. To study the actin filaments in these fluorescent regions more clearly, the soluble components of the cells were eliminated by treatment with Triton X-100 or saponin solution containing a low concentration of glutaraldehyde, and the detergent-treated and saponin-treated cells were observed under a transmission electron microscope. Concentrated actin filaments were observed directly beneath the Con A-induced capping area and CB-induced zeiotic knob-accumulation area. The area of concentrated actin filaments appeared to correspond to the electron dense area observed in the identical region in the cells fixed without detergent treatment. More actin filaments were observed in CB-treated cells than in Con A-treated ones.

Topics: Actins; Animals; Carcinoma, Ehrlich Tumor; Concanavalin A; Cytochalasin B; Mice; Microscopy, Electron; Microscopy, Electron, Scanning; Microscopy, Fluorescence

The glycogen-containing ascites cell line was found to have a 3-5 times higher 5'-nucleotidase specific activity than the glycogen-free variant, resulting in different substrate affinity constants of Km = 0.14 mM and Km = 0.69 mM respectively. These activity differences were due to true 5'-nucleotidase as shown by its inactivation through specific inhibitors such as concanavalin A and alpha, beta-methylene adenosine diphosphate. Substrate specificity of the enzyme was similar in both cell lines, but differences were observed with respect to the pH optimum and stability.

Topics: 5'-Nucleotidase; Alkaline Phosphatase; Animals; Carcinoma, Ehrlich Tumor; Cell Fractionation; Cell Membrane; Concanavalin A; Glycogen; Isoenzymes; Kinetics; Male; Mice; Mice, Inbred Strains; Nucleotidases; Substrate Specificity; Temperature

Alteration of surface receptor of Ehrlich tumour cells by concanavalin A (Con A), shortwave UV and UV plus Con A have been studied. DNA of nuclear chromatins is highly sensitive to UV but receptor glycoprotein sites are UV resistant as evidenced by their retention of agglutination properties after radiation. Con A and UV plus Con A treatments of tumour cells delay the in vivo growth of solid tumour which may be due to some immunological mechanism.

Topics: Agglutination Tests; Animals; Carcinoma, Ehrlich Tumor; Cell Survival; Cells, Cultured; Combined Modality Therapy; Concanavalin A; Dose-Response Relationship, Radiation; Male; Mice; Neoplasm Transplantation; Platelet Glycoprotein GPIb-IX Complex; Platelet Membrane Glycoproteins; Receptors, Cell Surface; Ultraviolet Therapy

Phytohemagglutinin (PHA) and concanavalin A (Con A) were used as probes to detect changes in the cell surface of Dalton's lymphoma, sarcoma-180 and Ehrlich's carcinoma after short in vitro exposure to acriflavine. Dye-treated cells showed enhancement of agglutination both by PHA and Con A, and such enhancement was found to be dependent on the time of exposure and concentration of acriflavine. However, PHA-induced percent agglutination seemed to be much higher than that of Con A among the 3 cell types. There were also marked differences among the 3 cell types in order of their sensitivity to lectin-mediated agglutination. The strength of the response was greater in lymphoma to both PHA and Con A than that of sarcoma-180 and carcinoma cells, which appeared to be most resistant. Acriflavine, which is known as an intercalative agent with DNA, induces cell surface changes by promoting lectin-mediated cellular agglutination.

Topics: Acriflavine; Agglutination; Aminoacridines; Animals; Carcinoma, Ehrlich Tumor; Cell Membrane; Cells, Cultured; Concanavalin A; Lymphoma; Male; Mice; Phytohemagglutinins; Sarcoma 180

Topics: Animals; Carcinoma, Ehrlich Tumor; Cations; Cell Membrane; Concanavalin A; Cytochalasin B; Cytoplasm; Cytoskeleton; Ferritins; Mice; Microscopy, Electron; Microvilli

Cytochalasin B (CB) treatment induces or accelerates the capping phenomenon in some cells. In Ehrlich ascites tumor cells (EATC) CB treatment apparently induced the capping of Con A binding sites as observed under a fluorescent microscope. However, electron microscopic examinations revealed that the CB treatment did not induce a rearrangement of Con A binding sites, but rather it only induced a change in cell shape. On the contrary, CB treatment inhibited the capping phenomenon induced by treatment with Con A. Electron microscopic observations may give exact information on the distribution of lectin binding sites.

Topics: Animals; Binding Sites; Carcinoma, Ehrlich Tumor; Cell Transformation, Neoplastic; Concanavalin A; Cytochalasin B; Immunologic Capping; Mice

In order to get precise information about the movement of plasma membrane proteins in cap formation, cyto- and bio-chemical analyses were made of the plasma membranes from non-capped areas of Ehrlich ascites tumor cells (EATCs) exposed to concanavalin A (Con A). Blebs formed by treatment with cytochalasin B (CB) of the non-capped areas of cells having a cap were isolated and used as the plasma membranes from non-capped areas (ConA-CB-bleb fraction). This bleb fraction was compared with a bleb fraction prepared from cells without ConA-treatment (CB-bleb fraction). Cytochemical analysis of ConA-CB-bleb fraction revealed a decreased in conA binding sites (ConA-BS) compared to the CB-bleb fraction. SDS polyacrylamide slab gel electrophoresis also revealed a decrease in the major components of ConA-BS of the ConA-CB-bleb fraction. The minor components of ConA-BS showed no distinct quantitative difference between the ConA-CB-bleb and CB-bleb fractions. NA+ K+-adenosine triphosphatase (ATPase), 5' nucleotidase (5'ND) and gamma-glutamyl transpeptidase (gamma-GTP) did not show any decrease in activity in the ConA-CB-bleb fraction, but the activity of D+-stimulated phosphatase (K-Pase) was decreased. The findings indicate that there are two types of plasma membrane glycoproteins in EATCs; one includes those participating in cap formation due to ConA, e.g. the major components of ConA-BS and K-Pase, and the other, those not participating in such cap formation, e.g. some minor components of ConA-BS, ATPase, 5'ND and gamma-GTP, which keep their places without moving.

Topics: 5'-Nucleotidase; Animals; Carcinoma, Ehrlich Tumor; Cell Membrane; Concanavalin A; Cytochalasin B; gamma-Glutamyltransferase; Histocytochemistry; Membrane Proteins; Mice; Microscopy, Electron; Nucleotidases; Phosphoric Monoester Hydrolases; Potassium; Sodium-Potassium-Exchanging ATPase

Trypsinized human erythrocytes were incubated with concanavalin A at 4 degrees C. After removal of free concanavalin A, the erythrocytes were incubated with Ehrlich ascites tumor cells at 37 degrees C. The erythrocytes formed rosettes with the tumor cells.

Topics: Animals; Carcinoma, Ehrlich Tumor; Concanavalin A; Erythrocytes; Humans; Rosette Formation; Trypsin

Cholinesterase activity was detected in the ascitic fluid of Ehrlich tumour and studied in a comparative manner in relation to that found in mice plasma. Enzymes from both sources were characterized with respect to optimum pH, substrate concentration and quinidine inhibition. After gel filtration by Sephadex G-200 and Sepharose 6B, two enzyme forms were observed in ascitic fluid as well as in mice plasma: a large form (L) and a small form (S) presenting molecular weights of 191 000, and 224 000 daltons for L forms and 71 000 and 69 000 daltons for S forms respectively. Concanavalin A interacts with both molecular forms, suggesting a glycoprotein nature for these enzymes.

Topics: Animals; Carcinoma, Ehrlich Tumor; Cholinesterases; Concanavalin A; Glycoproteins; Isoenzymes; Kinetics; Mice; Molecular Weight; Quinidine

Treatment of transformed Py3T3, SV101-3T3, and L1210 cells, as well as mitotic and Pronase-treated untransformed 3T3 cells, with the polyene antibiotics filipin, nystatin, and amphotericin B inhibited agglutination by wheat germ agglutinin. The effect of polyene antibiotic treatment was lectin and cell specific. Concanavalin A induced agglutination was not inhibited, wheat germ agglutination induced agglutination of untransformed 3T3 interphase cells was not influenced, and other aggregation phenomena, including those of erythrocytes with blood group specific antibodies or divalent cations, were unaffected by polyene treatments. This suggests that the formation of polyene-cholesterol complexes in transformed and erythrocyte cell membranes may specifically affect wheat germ agglutinin receptors and/or secondary events necessary for wheat germ agglutinin induced agglutination. Fluorescence studies of membrane filipin-cholesterol complexes showed that pretreating the cells with wheat germ agglutinin, but not concanavalin A, perturbed the fluorescence properties of filipin. Electron spin resonance studies with spin-labeled fatty acids revealed at best only a slight decrease in fatty acyl chain flexibility following filipin treatment. These studies indicate that there are not only quantitative differences between the agglutinability of transformed and untransformed cells with wheat germ agglutinin but that qualitative differences exist as well.

Topics: Agglutination; Amphotericin B; Animals; Carcinoma, Ehrlich Tumor; Cell Transformation, Viral; Cells, Cultured; Concanavalin A; Fibroblasts; Filipin; Lectins; Leukemia L1210; Mice; Mice, Inbred Strains; Mitosis; Nystatin; Polyenes; Structure-Activity Relationship

Topics: Agglutination Tests; Animals; Antibodies, Bacterial; Carcinoma, Ehrlich Tumor; Concanavalin A; Coombs Test; Cytotoxicity, Immunologic; Erythrocytes; Humans; Leukocytes; Micrococcus; Phytohemagglutinins

More than half of the DNA polymerase beta in mouse ascites cell chromatin was found to be associated with monomeric nucleosomal particles (produced by micrococcal nuclease treatment of chromatin). Almost all nuclear DNA polymerase activity in lymphocytes was found to be associated with nucleosomes. The nucleosome-associated enzyme was mainly DNA polymerase beta in chromatin from resting and mainly DNA polymerase alpha in chromatin from concanavalin-A-stimulated lymphocytes.

Topics: Animals; Carcinoma, Ehrlich Tumor; Chromatin; Concanavalin A; Deoxyribonucleases; DNA-Directed DNA Polymerase; Ethylmaleimide; Histones; Lymphocytes; Sodium Chloride

Topics: Animals; Carcinoma, Ehrlich Tumor; Concanavalin A; Female; Hemagglutination; Male; Mice; Mice, Inbred Strains; Neoplasm Transplantation; Protease Inhibitors; Transplantation, Homologous; Trypsin; Trypsin Inhibitors; Urokinase-Type Plasminogen Activator

Topics: Animals; Antineoplastic Agents; Anura; Capillary Permeability; Carcinoma, Ehrlich Tumor; Concanavalin A; Female; Lectins; Male; Mice; Ovum; Phagocytosis; Rana catesbeiana; Ranidae

The mRNA for the mouse MOPC-46B K chain, a glycorportein, was prepared translated in Ehrlich ascites cell-free extract. A polypeptide product similar to the MOPC-46B kappa chain was identified by its antigenic properties and by analysis of the products of tryptic digestion. In addition, the cell-free product was glycosylated after translation. It appeared that glycosylation of the cell-free product occurred on the same tryptic peptide glycosylated by intact cells. The characteristics and requirements of glycosylation are described. The presence of Mn2+ was a major requirement.

Topics: Animals; Antibody Formation; Carcinoma, Ehrlich Tumor; Cations, Divalent; Cell-Free System; Concanavalin A; Edetic Acid; Electrophoresis, Polyacrylamide Gel; Glycosides; Immune Sera; Immunoglobulin kappa-Chains; Immunoglobulin Light Chains; Lipids; Mannose; Mice; Neoplasm Proteins; Peptides; Plasmacytoma; Polyethylene Glycols; Protein Biosynthesis; Ribonucleases; RNA, Messenger; RNA, Neoplasm; Time Factors

Topics: Animals; Carcinoma, Ehrlich Tumor; Concanavalin A; Isotope Labeling; Mice; Technetium; Time Factors; Tissue Distribution

Topics: Animals; Carcinoma, Ehrlich Tumor; Cell Aggregation; Concanavalin A; Daunorubicin; Drug Combinations; Hydrocortisone; Leukemia L1210; Mice

Topics: Animals; Antibodies, Bacterial; Binding Sites; Blood Cells; Carcinoma, Ehrlich Tumor; Cell Aggregation; Concanavalin A; Erythrocytes; In Vitro Techniques; Lectins; Leukocytes; Micrococcus

Various reagents used in the chemical modification of amino- and carboxy-groups of proteins, and of carbohydrates of glycoproteins and glycolipids, inhibit respiration in ascites tumor cells concomitant with release of potassium ion from those cells. The respiratory activity of washed ascites tumor cells is increased by exogenous addition of potassium ion. The lowered respiratory control index as well as oxidative phosphorylation of aged mitochondria are restored upon increasing the potassium concentration of the incubation mixture in the presence of respiratory substrates. The data suggest that the potassium ion level of cells is changed by modifying physicochemical properties of membrane components and that cellular energy metabolism is regulated by intracellular potassium ion concentration.

Topics: 4-Aminobenzoic Acid; Acrolein; Animals; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Cell Membrane; Cells, Cultured; Cobalt; Concanavalin A; Dansyl Compounds; Dicyclohexylcarbodiimide; In Vitro Techniques; Lanthanum; Liver Neoplasms; Mitochondria, Liver; Ouabain; Oxidative Phosphorylation; Oxygen Consumption; Potassium; Rats; Ruthenium Red; Trinitrobenzenesulfonic Acid

Concanavalin A (Con A)-binding glycoproteins have been purified from Ehrlich ascites tumor cells by precipitation of cell extracts with Con A in agarose gels and by lithium diiodosalicylate (LIS) extraction. Antisera prepared against these two glycoprotein fractions indicate the presence of a minimum of five different antigens in the Con A-cell extract precipitate and a minimum of three different antigens in the LIS-extracted preparation. That both kinds of antisera are directed towards surface membrane antigens is strongly suggested by the fact that they give precipitin bands with purified plasma membrane from Ehrlich cells, they agglutinate whole cells, and they no longer give precipitin bands after absorption by whole cells. Cell extract material which was specifically eluted from a Con A-Sepharose 4B column with alpha-methylmannoside was recognized by antisera to both glycoprotein preparation, demonstrating that the antigens were Con A-binding components; material which did not bind the column failed to react with either antisera. Immunochemical analysis indicated that one of the antigens present in the LIS-extracted material was distinct from the five antigens prepared by lectin precipitation in agarose gels suggesting that a minimum of six individual Con A receptors are present on the surface of Ehrlich tumor cells. Immunochromatography was found to be a convenient method to purify the antigens obtained by the LIS extraction of Ehrlich cells.

Topics: Animals; Binding Sites; Binding Sites, Antibody; Carcinoma, Ehrlich Tumor; Cell Membrane; Concanavalin A; Glycoproteins; Immunodiffusion; Immunoelectrophoresis; Male; Mice; Molecular Weight; Protein Binding; Rabbits; Receptors, Drug

Investigations on methods for the utilization of fluorescein-isothiocyanate-labelled Concanavalin A, Lens culinaris lectin and Ricinus communis lectin for immunohistological demonstration as simple sugar moieties are reported. The stainings were carried out on rabbit erythrocytes. Ehrlich ascitee tumor cells and various mammalian tissues. Optimum results were obtained in living cells and native cryostate tissue sections. The influence of different fixing agents and conditions of fixation on the degree of tissue flurorescence were studied. Generally the fluorescense decreased by aldehyde fixation. Similar effect was observed following fixation by heat. Furthermore, changes in the pattern of fluorescence depending on the type of tissue and utilized lectin were observed after aldehyde fixation. The degree of tissue fluorescence was fairly independent of the pH value. The specificity of the particular reactions for saccharide demonstration was shown; glycerol is capable of a differently strong "hapten-like" action.

Topics: Animals; Carbohydrates; Carcinoma, Ehrlich Tumor; Concanavalin A; Erythrocytes; Histocytochemistry; Lectins; Methods; Mice; Plant Lectins; Plants; Plants, Toxic; Rabbits; Rats; Ricinus; Staining and Labeling; Thiocyanates

Topics: Animals; Antigen-Antibody Reactions; Carcinoma, Ehrlich Tumor; Cell Membrane; Concanavalin A; Ferritins; Freeze Etching; Receptors, Concanavalin A; Receptors, Drug

Topics: Agglutination; Animals; Carcinoma, Ehrlich Tumor; Cell Line; Concanavalin A; Potassium; Temperature

Topics: Agglutination; Animals; Azides; Binding Sites; Carcinoma, Ehrlich Tumor; Cell Membrane; Colchicine; Concanavalin A; Depression, Chemical; Dinitrophenols; Erythrocytes; Hydroxides; Iodine Radioisotopes; Iodoacetates; Papain; Periodic Acid; Phenylhydrazines; Receptors, Drug; Rotenone

Topics: Animals; Carbon Radioisotopes; Carcinoma, Ehrlich Tumor; Cell Adhesion; Cell Aggregation; Cells, Cultured; Chick Embryo; Concanavalin A; Haptens; Macromolecular Substances; Methylglucosides; Muscles; Receptors, Drug; Retina; Trypsin

A report is given about a manyside applicable method to examine the agglutination of tumour cells and normal cells. It enables a presentation of results without subjective influences and is based on the electronic determination of the diminution of impulse numbers with a particle counter. This method is useful with little alterations for example to determine sugar specificity of lectins, effectivity of cytostatics and antisera against tumour cells and appropriate healthy cells.

Topics: Animals; Carbohydrates; Carcinoma, Ehrlich Tumor; Cell Aggregation; Concanavalin A; Female; Galactose; Glucosamine; Glucose; Lectins; Methods; Mice; Rats; Thymus Gland

A correlation has been observed between concanavalin A agglutination of various cell types and the presence of surface membrane galactosyltransferase (1-O-alpha-D-Galactosyl-myo-inositol:raffinose galactosyltransferase, EC 2.4.1.67) activity. Moreover, a reduction to less than 50% of cell surface galactosyltransferase activity occurred after treatment with concanavalin A; other cell surface glycosyltransferase enzyme activities examined were unaffected by concanavalin A treatment. To confirm the participation of cell surface galactosyltransferase in concanavalin A-induced cell agglutination, the enzyme from rabbit erythrocytes was solubilized by sonication and purified by preparative polyacrylamide gel electrophoresis. It was possible to achieve a purified preparation of rabbit erythrocyte galactosyltransferase by separation on concanavalin A-Sepharose. The purified enzyme showed visible immunoprecipitation (Ouchterlony) with concanavalin A. Furthermore, human erythrocytes, which are not normally agglutinated by concanavalin A, became agglutinable by the lectin when the erythrocytes were preincubated with purified galactosyltransferase. These experiments suggest a direct and possible specific role of cell surface galactosyltransferase enzyme in the mechanism of concanavalin A agglutination of cells.

Topics: Agglutination; Animals; Antigen-Antibody Reactions; Carcinoma, Ehrlich Tumor; Cell Fractionation; Cell Line; Cell Membrane; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Erythrocytes; Galactose; Hemagglutination; Hexosyltransferases; Humans; Immunoglobulin M; Intestines; Precipitin Tests; Rabbits; Rats; Species Specificity

Topics: Animals; Binding Sites, Antibody; Carcinoma, Ehrlich Tumor; Cell Membrane; Concanavalin A; Ferritins; Fluoresceins; Fluorescent Antibody Technique; Freeze Etching; Mice; Microscopy, Electron; Peroxidases; Thiocyanates

Topics: Agglutination; Animals; Biological Transport, Active; Carcinoma, Ehrlich Tumor; Cells, Cultured; Chlorides; Concanavalin A; Glycine max; Lectins; Mice; Ouabain; Plant Lectins; Potassium; Sodium; Time Factors

Topics: Agglutination; Animals; Binding Sites, Antibody; Carcinoma, Ehrlich Tumor; Concanavalin A; Culture Techniques; Formaldehyde; Hemadsorption; Hemagglutination Tests; Iodine Radioisotopes; Orthomyxoviridae; Virus Replication

Topics: Animals; Carcinoma, Basal Cell; Carcinoma, Ehrlich Tumor; Cell Adhesion; Cell Fusion; Cells, Cultured; Concanavalin A; Humans; Macrophages; Mice; Microscopy, Electron; Phagocytosis; Radiation Effects; Skin; Skin Neoplasms; Staining and Labeling; Time Factors

Topics: Agglutination; Animals; Azides; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Cell Membrane; Cells; Concanavalin A; Cycloheximide; Cytochalasin B; Dimethyl Sulfoxide; Dinitrophenols; Glycosides; Lectins; Liver Neoplasms; Rats; Sarcoma, Yoshida; Temperature; Tritium

Topics: Agglutination; Animals; Carcinoma, Ehrlich Tumor; Cell Aggregation; Cell Line; Cells; Concanavalin A; Cricetinae; In Vitro Techniques; Lectins; Trypsin Inhibitors

Topics: Agglutination; Animals; Carcinoma, Ehrlich Tumor; Cells, Cultured; Concanavalin A; Galactose; Hydrogen-Ion Concentration; In Vitro Techniques; Iodine Radioisotopes; Lactose; Neoplasm Proteins; Plant Lectins; Plants, Toxic; Protein Binding; Ricin; Ricinus; Serum Albumin, Bovine; Toxins, Biological

Topics: Adenosine Triphosphatases; Animals; Biphenyl Compounds; Carcinoma, Ehrlich Tumor; Cell Membrane; Concanavalin A; Histocytochemistry; Macrophages; Male; Mice; Microscopy, Electron; Peritoneum; Peroxidases; Phagocytosis; Radiation Effects; Staining and Labeling

Concanavalin A (Con-A) was linked to ferritin with glutaraldehyde and chromatographed on Sepharose 6B to separate unconjugated Con-A and ferritin from covalently cross-linked molecules. Ehrlich ascites tumor cells were infected with WSA influenza virus, stained at intervals with the ferritin-labeled Con-A and examined by electron microscopy. The surfaces of most mature viruses were specifically stained, providing direct evidence that influenza viruses maturing in this cell type have exposed Con-A receptor sites. The ferritin cores of the staining reagent were found at an average distance of 21.3 nm from the virus membrane and 10.8 nm from the uninfected cell membrane. This finding was interpreted to mean that the population of Con-A receptor sites on influenza virus particles is located at an average distance from the virus membrane twice that of the population of Con-A receptor sites found on uninfected cells. The structural elements of viral membranes can provide a reliable means for evaluating electron microscopy staining reagents, thereby enhancing their usefulness as probes for the study of membrane relationships.

Topics: Animals; Binding Sites; Carcinoma, Ehrlich Tumor; Cell Membrane; Chromatography; Concanavalin A; Erythrocytes; Ferritins; Fluorescent Antibody Technique; Hemagglutination Tests; Hemagglutination, Viral; Lectins; Mice; Microscopy, Electron; Microscopy, Fluorescence; Orthomyxoviridae; Protein Binding; Temperature

Topics: Agglutination; Animals; Carcinoma, Ehrlich Tumor; Cell Adhesion; Concanavalin A; Glucose; Glycosides; In Vitro Techniques; Lectins; Macrophages; Mice; Neoplasm Transplantation; Peritoneal Cavity; Phagocytosis

Topics: Animals; Carbohydrate Metabolism; Carcinoma, Ehrlich Tumor; Cell Membrane; Concanavalin A; Histocytochemistry; In Vitro Techniques; Lectins; Leukemia, Experimental; Methods; Mice; Microscopy, Electron

Topics: Animals; Ascitic Fluid; Carcinoma, Ehrlich Tumor; Cattle; Chickens; Circular Dichroism; Concanavalin A; Erythrocytes; Glucose; Guinea Pigs; Lectins; Macrophages; Mannose; Mice; Microscopy, Electron; Phagocytosis; Rabbits; Rats