concanavalin-a and Cell-Transformation--Neoplastic

Heterogeneous distribution of surface domains is a characteristic feature of the tumor cell surface and the distribution differs from that of normal cells. During the malignant transformation the heterogeneity may change or disappear. Cell lines with various metastasizing capacities show different distributions of membrane domains or other differences in membrane or surface organization. We have demonstrated that the amount and distribution of negatively charged sites of B 16 melanoma membranes changed upon ionizing radiation (X-ray, 60Co-gamma). In the case of the P 388 lymphoma, however, only the amount of negatively charged sites change after irradiation, the distribution remains unaltered. Both features proved to be radioresistant in human lymphoid leukemic cells.

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Ferritins; Histocytochemistry; Humans; Leukemia, Lymphoid; Lymphoma; Male; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Microscopy, Electron; Protein Binding; Tumor Cells, Cultured

Topics: Animals; Cell Adhesion; Cell Communication; Cell Differentiation; Cell Division; Cell Membrane; Cell Movement; Cell Transformation, Neoplastic; Concanavalin A; Cytoplasm; Cytoskeleton; Humans; In Vitro Techniques; Lymphocyte Activation; Microtubules; Models, Biological; Receptors, Concanavalin A; Receptors, Drug

Topics: Agglutination; Blood Group Antigens; Cell Membrane; Cell Transformation, Neoplastic; Cell Transformation, Viral; Chemical Phenomena; Chemistry; Concanavalin A; Female; Lectins; Lymphocyte Activation; Male; Microvilli; Mitogens; Neoplasms, Experimental; Ovum; Receptors, Drug; Spermatozoa; Zygote

Topics: Agammaglobulinemia; Animals; Antigens; Autoimmune Diseases; Binding, Competitive; Cell Transformation, Neoplastic; Chickens; Concanavalin A; Dysgammaglobulinemia; Epitopes; Genes; Humans; Immune Tolerance; Immunity, Cellular; Immunoglobulin A; Immunoglobulin Allotypes; Immunoglobulin E; Immunologic Deficiency Syndromes; Immunosuppression Therapy; Lymphokines; Mice; Mycoses; Rabbits; T-Lymphocytes

Topics: Adenine; Animals; Binding Sites; Biological Transport; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Fibroblasts; Leukocytes; Lysine; Macrophages; Microscopy, Electron; Microtubules; Models, Biological; Phagocytosis; Rabbits; Simian virus 40

Infection of untransformed cells with a wide-range of non-oncogenic enveloped viruses causes a significant increase in their susceptibility to agglutination by concanavalin A (Con A). The increased Con A agglutinability of these cells is not caused by an increase in the number of Con A sites on the cell surface but involves alteration in the surface properties of infected cells to allow redistribution of Con A receptors to form "patches" following binding of Con A to the cell surface. Similarities between Con A-mediated agglutination of normal cells infected with non-oncogenic viruses and the agglutination response to cells transformed by oncogenic viruses will be reviewed. Finally, the use of Con A as an experimental tool to modify the replication and cytopathogenicity of non-oncogenic viruses grown in mammalian cells will be presented.

Topics: Agglutination; Animals; Binding Sites; Cattle; Cell Aggregation; Cell Fusion; Cell Transformation, Neoplastic; Cells; Chick Embryo; Concanavalin A; Cricetinae; Cytopathogenic Effect, Viral; Enzymes; Newcastle disease virus; Virus Replication; Viruses

Topics: Abrin; Agglutination; Animals; Antineoplastic Agents; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Glycoproteins; Hodgkin Disease; Humans; Immunity, Cellular; In Vitro Techniques; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Neoplasms; Receptors, Drug; Ricin

Topics: Agglutination Tests; Animals; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fibroblasts; Glucose; Glycosides; Haptens; Lectins; Leukemia L1210; Lipase; Mannose; Mice; Mice, Inbred Strains; Mitosis; Models, Biological; Peptide Hydrolases; Polyomavirus; Time Factors

Topics: Adrenocorticotropic Hormone; Animals; B-Lymphocytes; Bucladesine; Cell Transformation, Neoplastic; Concanavalin A; Cyclic AMP; Cyclic GMP; Humans; Immunity; Kinetics; Lectins; Lymphocyte Activation; Lymphocytes; Models, Biological; Phagocytosis; Subcellular Fractions; T-Lymphocytes; Theophylline

Topics: Animals; Binding Sites; Carcinogens; Cell Adhesion; Cell Differentiation; Cell Membrane; Cell Transformation, Neoplastic; Chromosomes; Concanavalin A; Fibroblasts; Granulocytes; Hematopoietic Stem Cells; Lectins; Leukemia; Leukemia, Experimental; Leukemia, Myeloid; Macrophages; Models, Biological; Neoplasms; Neoplasms, Experimental; Phenotype; Polyomavirus; Simian virus 40

Topics: Agglutination; Animals; Binding Sites; Cell Division; Cell Membrane; Cell Transformation, Neoplastic; Chemical Phenomena; Chemistry; Chromatography, Affinity; Concanavalin A; Erythrocytes; Glycine max; Glycoproteins; Lectins; Lymphocytes; Membranes; Mitogens; Molecular Conformation; Plant Lectins; Plants, Toxic; Polysaccharides; Receptors, Drug; Ricinus communis; Surface Properties; Triticum; Viruses

Topics: Agglutination; Animals; Antigens; Bacteria; Binding Sites; Biological Evolution; Cattle; Cell Membrane; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Female; Lectins; Maternal-Fetal Exchange; Oncogenic Viruses; Parasites; Pregnancy; Vinblastine

Topics: Agglutination; Agglutination Tests; Animals; Avian Sarcoma Viruses; Binding Sites; Binding, Competitive; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Concanavalin A; Cricetinae; Cyclic AMP; Fibroblasts; Glycolipids; Kidney; Lectins; Mice; Mitosis; Models, Biological; Peptide Hydrolases; Pinocytosis; Polyomavirus; Receptors, Drug

Topics: Adenylyl Cyclases; Adipose Tissue; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Humans; Insulin; Insulin Resistance; Iodine Isotopes; Liver; Mitogens; Neuraminidase; Receptors, Cell Surface

Topics: Adenylyl Cyclases; Animals; Bucladesine; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Contact Inhibition; Culture Media; Cyclic AMP; DNA; Fibroblasts; Growth Substances; HeLa Cells; Humans; Isoproterenol; L Cells; Lectins; Lymphocytes; Mammals; Mitogens; Mitosis; Neoplasms; Prostaglandins; Protein Precursors; RNA; Theophylline

Topics: Cell Transformation, Neoplastic; Chemical Phenomena; Chemistry, Physical; Concanavalin A; Glycine max; Glycoproteins; Lectins; Lymphocyte Activation; Lymphocytes; Methods; Plant Lectins; Polysaccharides; Receptors, Drug

Chronic inflammation is strongly associated with an increased risk for hepatocellular carcinoma (HCC) development. The multidrug resistance 2 (Mdr2)-knockout (KO) mouse (adenosine triphosphate-binding cassette b4(-/-) ), a model of inflammation-mediated HCC, develops chronic cholestatic hepatitis at an early age and HCC at an adult age. To delineate factors contributing to hepatocarcinogenesis, we compared the severity of early chronic hepatitis and late HCC development in two Mdr2-KO strains: Friend virus B-type/N (FVB) and C57 black 6 (B6). We demonstrated that hepatocarcinogenesis was significantly less efficient in the Mdr2-KO/B6 mice versus the Mdr2-KO/FVB mice; this difference was more prominent in males. Chronic hepatitis in the Mdr2-KO/B6 males was more severe at 1 month of age but was less severe at 3 months of age in comparison with age-matched Mdr2-KO/FVB males. A comparative genome-scale gene expression analysis of male livers of both strains at 3 months of age revealed both common and strain-specific aberrantly expressed genes, including genes associated with the regulation of inflammation, the response to oxidative stress, and lipid metabolism. One of these regulators, galectin-1 (Gal-1), possesses both anti-inflammatory and protumorigenic activities. To study its regulatory role in the liver, we transferred the Gal-1-KO mutation (lectin galactoside-binding soluble 1(-/-) ) from the B6 strain to the FVB strain, and we demonstrated that endogenous Gal-1 protected the liver against concanavalin A-induced hepatitis with the B6 genetic background but not the FVB genetic background.. Decreased chronic hepatitis in Mdr2-KO/B6 mice at the age of 3 months correlated with a significant retardation of liver tumor development in this strain versus the Mdr2-KO/FVB strain. We found candidate factors that may determine strain-specific differences in the course of chronic hepatitis and HCC development in the Mdr2-KO model, including inefficient anti-inflammatory activity of the endogenous lectin Gal-1 in the FVB strain.

Topics: Animals; ATP Binding Cassette Transporter, Subfamily B; ATP-Binding Cassette Sub-Family B Member 4; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; Chemical and Drug Induced Liver Injury; Concanavalin A; Galectin 1; Hepatitis, Chronic; Liver; Liver Neoplasms; Male; Methionine Adenosyltransferase; Mice; Mice, Inbred Strains; Mice, Knockout

Beta-endorphin and the synthetic beta-endorphin-like decapeptide Ser-Leu-Thr-Cys-Leu-Val-Lys-Gly-Phe-Tyr (referred to as immunorphin), corresponding to the sequence 364-373 of the CH3 domain of human immunoglobulin G heavy chain, were shown to stimulate concanavalin A-induced proliferation of T lymphocytes from the blood of healthy donors. [Met(5)]Enkephalin and the antagonist of opioid receptors naloxone examined in parallel were inactive. The stimulating effect of beta-endorphin and immunorphin on T lymphocyte proliferation is not inhibited by naloxone. Studies on receptor binding of (125)I-labeled immunorphin to T lymphocytes revealed that it binds with high affinity to naloxone-insensitive receptors (K(d) = 7.0 +/- 0.3 nM). Unlabeled immunorphin completely inhibits (125)I-labeled beta-endorphin specific binding to naloxone-insensitive receptors on T lymphocytes (K(i) = 0.6 +/- 0.1 nM). Thus, beta-endorphin and immunorphin interact with common naloxone-insensitive receptors on T lymphocytes.

Topics: beta-Endorphin; Binding, Competitive; Cell Division; Cell Transformation, Neoplastic; Concanavalin A; Enkephalin, Methionine; Humans; Immunoglobulin Constant Regions; Immunoglobulin G; Immunoglobulin gamma-Chains; Immunoglobulin Heavy Chains; Iodine Radioisotopes; Lymphocyte Activation; Naloxone; Narcotic Antagonists; Oligopeptides; Peptide Fragments; Receptors, Opioid; T-Lymphocytes

Mucinous tumors of the ovary have been thought to originate in two ways: by müllerian-type metaplasia of surface epithelium, and as monodermal teratomas. To gain a better understanding of their pathogenesis, the authors analyzed these tumors for their expression of estrogen receptors (ER) and progesterone receptors (PR) as markers of müllerian-type differentiation and for their content of gastric-type mucin as a marker of gastric differentiation.. The histochemical expression of ER, PR, and gastric-type mucin was studied in 10 specimens of the cervix with normal endocervical glands (as a representative of müllerian-derived mucin-containing cells), 3 ovary specimens with surface epithelial inclusion cysts that contained endocervical-like mucin-containing cells (representing müllerian-type metaplasia), and 47 mucinous tumors of the ovary (29 benign, 8 with low malignant potential, and 10 malignant).. Normal endocervical glands expressed ER and PR and rarely expressed gastric-type mucin. Ovarian inclusion cysts showed strong expression of ER and PR in the cuboidal cells and drastically reduced expression in the endocervical-like mucin-containing cells. The cuboidal cells were negative for gastric-type mucin, but the endocervical-like mucin-containing cells expressed gastric-type mucin. Endocervical-like mucinous cells in benign and borderline mucinous tumors showed expression of PR and/or gastric-type mucin in all cases.. The staining results for the inclusion cysts support the thesis that the endocervical-like mucinous cells encountered in the ones that express ER and PR weakly or not at all and have histochemical properties of normal gastric epithelium have their origin in metaplasia of müllerian-type epithelium. Application of the same staining methods to benign ovarian tumors and those with low malignant potential suggests strongly that similar müllerian-type metaplasia is a major pathway in their pathogenesis.

Topics: Adenocarcinoma, Mucinous; Adult; Aged; Cell Differentiation; Cell Transformation, Neoplastic; Cervix Uteri; Concanavalin A; Cystadenoma, Mucinous; Female; Galactose Oxidase; Gastric Mucins; Humans; Immunohistochemistry; Metaplasia; Middle Aged; Mullerian Ducts; Ovarian Cysts; Ovarian Neoplasms; Receptors, Estrogen; Receptors, Progesterone; Staining and Labeling

We examined the functional consequences of cellular transformation of rat IAR-2 epithelial cells, by a mutant N-ras oncogene, on the dynamics of active lamellae, structures that play an important role in cell motility, adhesion, and surface-receptor capping. Lamellar activity was assessed by measuring the rate of outer-edge pseudopodial activity and by analyzing the motility of Con A-coated beads placed on lamellar surfaces with optical tweezers. Although transformation dramatically affected the shape and size of active cellular lamellae, there was little detectable effect on either pseudopodial activity or bead movement. To investigate the potential relationship between functional lamellar activity and the microtubule cytoskeleton, lamellar activity was examined in nontransformed and transformed cells treated with the microtubule-disrupting drug nocodazole. In the absence of microtubules, transformed cells were less polarized and possessed decreased rates of pseudopodial and bead motility. On the basis of these observations, it is suggested that ras-induced transformation of epithelial cells consists of two cytoskeletal modifications: overall diminished actin cytoskeletal dynamics in lamellae and reorganization of the microtubule cytoskeleton that directs pseudopodial activity to smaller polarized lamellae.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cytoskeleton; Epithelium; Gene Expression Regulation; Genes, ras; Liver; Microtubules; Nocodazole; Pseudopodia; Rats

Authors examined 18 cases of osteoclastoma chosen at random with different lectins. The fibroblast-like type I stromal cells taking part in the formation of the ground substance were Con A positive. The strength of the Con A bond and the number of positive cells increased with the progress of the stage. The PNA lectin bond following the digestion with neuraminidase was in the single stages of different character and intensity. Whereas in the Gr I osteeoclastomas 30 per cent of the reacting osteoclast was localized to a well defined segment of the membrane in the Gr III tumors the ratio of the negative cells moved around 1 per cent, and in 60 per cent of the positive cells strong diffuse intracytoplasmatic reaction could be observed. Authors think that the lectin histochemical examinations give help to the definition of the activity of the tumor forming cells.

Topics: Adult; Cell Transformation, Neoplastic; Concanavalin A; Female; Giant Cell Tumor of Bone; Histocytochemistry; Humans; Lectins; Male; Neoplasm Staging; Osteoclasts; Random Allocation

The binding pattern of two lectins, concanavalin A (ConA) and peanut agglutin (PNA), in various phases of tumour progression in the oral epithelium was studied. These included non-dysplastic, dysplastic and neoplastic lesions as well as normal tissue. ConA and PNA showed intense staining in the basement membrane of all types of lesions. Little difference was observed in the staining patterns between different stages of oral carcinogenesis, either with ConA or PNA. ConA showed mild cytoplasmic and membrane staining in all types of lesions while PNA showed moderate to intense staining in both the cytoplasm and membrane of lower-layer cells in all histological groups. The present study therefore shows that these lectins have limited value in the elucidation of oral carcinogenesis and are of insignificant diagnostic value.

Topics: Carcinoma; Carcinoma, Papillary; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Concanavalin A; Glycoconjugates; Humans; Lectins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Neoplasm Proteins; Peanut Agglutinin; Protein Binding

Blood monocytes (BMs) from 139 subjects (70 malignant melanoma patients, 31 breast cancer patients, 38 healthy controls) were cultured for at least 7 days. The formation of multinucleated giant cells (MGCs), which was checked during the whole time of culture, was observed in all cases. By the seventh day MGCs represented 25-50% and during the second and third month more than 90% of all cells. Lymphokines and/or concanavalin A stimulation (16-34 cases respectively) of BMs was performed as well. This stimulation greatly accelerated MGC formation. There were no differences either in spontaneous or in stimulated fusion between the different groups compared.

Topics: Breast Neoplasms; Cell Fusion; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Giant Cells; Humans; Lymphokines; Monocytes; Multiple Myeloma

Lectin binding to tumor cells in tissue sections of nonmetastatic and metastatic murine Lewis lung carcinoma (LLC) was assessed by light and electron microscopy using a lectin-gold technique. Ulex europaeus agglutinin-I (UEA-I) and peanut agglutinin (PNA) showed no binding, whereas concanavalin A (Con A), soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Maclura pomifera agglutinin (MPA), and Ricinus communis agglutinin-I (RCA-I) bound equally to the transplanted sites and metastases. However, wheat germ agglutinin (WGA) bound to metastases more highly than to the transplanted sites and there was a statistically significant difference (P less than 0.01) between the transplanted sites and metastases with regard to pre-embedding method. The tumor cells binding to WGA clearly decreased in number after sialic acid pretreatment and were rich in more well-differentiated organelle. In the bromodeoxyuridine (BrdUrd) labeling in vivo, cell proliferation was greater in the metastatic sites than in the transplanted sites. The above findings suggest that glycoconjugates on the tumor cell surface are altered in the process of metastasis and correlate with metastatic potential and cell proliferation.

Topics: Animals; Bromodeoxyuridine; Cell Count; Cell Division; Cell Transformation, Neoplastic; Concanavalin A; Histocytochemistry; Immunohistochemistry; Lectins; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Microscopy, Electron; Neoplasm Metastasis; Neoplasms, Experimental; Peanut Agglutinin; Plant Lectins; Tumor Cells, Cultured

Treatment of splenic leukocytes from Cornell K strain male chickens (homozygous at the B15 locus of the major histocompatibility complex) with ovine corticotropin-releasing factor (oCRF), before their co-incubation with naive chicken adrenal cells, resulted in an increase in corticosterone production. Supernatants from the oCRF-treated splenic leukocytes caused a time-dependent increase in corticosterone production when incubated with chicken adrenal cells. Adding oCRF directly to chicken adrenal cells did not increase corticosterone production. Pretreatment of peripheral leukocytes with oCRF increased their activity in a concanavalin A mitogen assay. Thus, chicken leukocytes stimulated with corticotropin releasing factor appear to increase the production of an "adrenocorticotropin-like" substance (adrenocorticotropin-like because it increases corticosterone production by adrenal cells), and increased their cell-mediated immune activity.

Topics: Adrenal Glands; Adrenocorticotropic Hormone; Animals; Biological Assay; Cell Transformation, Neoplastic; Chickens; Concanavalin A; Corticosterone; Corticotropin-Releasing Hormone; Dose-Response Relationship, Drug; In Vitro Techniques; Leukocytes; Male; Regression Analysis; Spleen; Time Factors

We examined the effects of pure first-trimester human trophoblast cells grown in long-term cultures or their secreted products on the proliferation of human peripheral blood lymphocytes cultured alone, under allogeneic stimulation, or in the presence of concanavalin A. Both trophoblasts and their culture supernatants stimulated lymphocyte proliferation. Culture supernatant had a moderate enhancing effect on lymphocyte mitogenesis in mixed lymphocyte cultures and in the presence of concanavalin A. Anti-human chorionic gonadotropin antibody suppressed the proliferative effect of trophoblast cells and their supernatants in the above experiments in a dose-dependent manner. At physiologic concentrations, both pure and impure forms of human chorionic gonadotropin enhanced (in a dose-dependent manner) proliferation of lymphocytes cultured alone (peak stimulation index, 6 to 7.8 at approximately 7 IU/ml). At higher concentrations (20 to 400 IU/ml) the proliferative effect was abolished. Trophoblast culture supernatant induced the expression of interleukin-2 receptors on lymphocytes after 48 hours of incubation. The supernatant also stimulated proliferation of human colon carcinoma cells. Thus trophoblast or trophoblast-derived human chorionic gonadotropin has a lymphocytotrophic function that may have implications for fetal survival.

Topics: Carcinoma; Cell Communication; Cell Division; Cell Line; Cell Transformation, Neoplastic; Chorionic Gonadotropin; Colonic Neoplasms; Concanavalin A; Culture Media; Female; Humans; Isoantigens; Lymphocyte Activation; Lymphocytes; Pregnancy; Pregnancy Trimester, First; Receptors, Interleukin-2; Trophoblasts

Transglutaminase (TGase) activity was reduced in intact mitogen-stimulated human peripheral blood lymphocytes (PBL) when compared to intact resting PBL. Moreover, a treatment of the same quiescent immunocompetent cells with purified liver TGase and Ca2+ completely suppressed the mitogen-induced blast transformation. A decrease in TGase activity in neoplastically transformed seminal vesicle epithelial cells with respect to their normal parent counterpart was also observed. Our data support the notion of a possible implication of TGase in cell proliferation and transformation.

Topics: Animals; Binding, Competitive; Calcium; Catalysis; Cell Division; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; Harvey murine sarcoma virus; Kirsten murine sarcoma virus; Liver; Lymphocytes; Mitogens; Proteins; Rats; Rats, Inbred Strains; Transglutaminases; Tumor Cells, Cultured

Protein phosphorylation mediated by murine IL3 and other factors has been studied in two different IL3-dependent lines, AC2 and 123. In both lines, responses to rat recombinant IL3 are enhanced or induced by growth in rat spleen lymphocyte conditioned medium. Growth stimulation by murine and rat IL3, by rat lymphokine(s), and by ATP in ATP-responsive cells is closely associated with the rapid (2-4 min) phosphorylation of a 33-kDa protein (p33) in all the cells examined. p33 phosphorylation is not stimulated by another lymphokine, IL4, nor by TPA or calcium ionophore alone, which are unable to stimulate growth by themselves, and is independent of serum. p33 phosphorylation is inhibited by trifluoperazine, an inhibitor of calcium-calmodulin, but is less sensitive to inhibition by H7, an inhibitor of protein kinase c, in AC2 cells. A spontaneous IL3-independent clone of AC2 (AC-) has been isolated. AC- cells are aggressively leukemic, do not produce detectable IL3, but phosphorylate p33 constitutively where it is associated with a particulate cell fraction. It is suggested that p33 is a common intermediate molecule involved in signal transduction by the various ligands which result in growth stimulation and that its constitutive phosphorylation may play a key role in the maintenance of the leukemic state.

Topics: Adenosine Triphosphate; Animals; Calcimycin; Cell Division; Cell Line; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Culture Media; Interleukin-3; Interleukin-4; Interleukins; Kinetics; Mice; Molecular Weight; Phosphoproteins; Phosphorylation; Protein Kinase C; Rats; Recombinant Proteins; Spleen; Tetradecanoylphorbol Acetate

Numatrin is a tightly bound nuclear matrix protein (40 kD/pI-5) whose synthesis is markedly and promptly increased in association with cellular commitment for mitogenesis in B lymphocytes. (Feuerstein, N., and J.J. Mond. 1987. J. Biol. Chem. 262:11389-11397). To study whether this event is exclusively associated with proliferation of B lymphocytes, we examined the synthesis of numatrin in T lymphocytes (murine and human) activated by lectins or by anti-T cell antigen receptor monoclonal antibody and in Swiss 3T3 fibroblasts stimulated by growth factors. We showed a close correlation between induction of DNA synthesis and induction of numatrin synthesis in T lymphocytes stimulated by concanavalin A, anti-T cell antigen receptor monoclonal antibody, and IL-2 in murine T cells. Similar results were observed in Swiss 3T3 fibroblasts, thus only combinations of growth factors (insulin/EGF or insulin/B subunit of cholera toxin) or serum, which induced a significant increase in DNA synthesis, were also associated with a significant increase in synthesis of numatrin. Similar to B cells, the increase in numatrin synthesis in fibroblasts was found to occur at early G1 phase. The calcium ionophores, A23187 and ionomycin, previously shown to induce an increase in c-myc and c-fos mRNA levels in fibroblasts, induced a marked increase in the synthesis of a nuclear protein at 80 kD/pI-5 but failed to induce an increase in the synthesis of numatrin indicating that an increase in intracellular Ca++ level is not sufficient for induction of the synthesis of numatrin. This further indicates that the increase in synthesis of numatrin may be more closely correlated with cellular commitment for mitogenesis as compared with other biochemical parameters. Using a polyclonal numatrin antibody we demonstrated that mitogen stimulation is also associated with a marked increase in numatrin abundance, which reached a peak at the onset of S phase and declined at the end of S phase. Evidence is presented to show that numatrin synthesis and abundance is elevated in various lymphoma cell lines. Using indirect immunofluorescence assays we showed that numatrin is abundant in other malignant cells: KB, epidermoid carcinoma, and Hep2 human hepatoma. Immunofluorescence studies further showed that mitogen stimulation of B lymphocytes induced a marked accumulation of numatrin in the nucleoli. This observation is in accord with the recent finding of identity of numatrin with the nucleolar protein B23

Topics: Antibodies, Monoclonal; B-Lymphocytes; Blotting, Western; Calcimycin; Cell Cycle; Cell Division; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; DNA; Electrophoresis, Gel, Two-Dimensional; Fluorescent Antibody Technique; Growth Substances; Humans; Interleukin-2; Kinetics; Nuclear Proteins; Nucleophosmin; Receptors, Antigen, T-Cell; T-Lymphocytes

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Clone Cells; Concanavalin A; DNA; Humans; Interleukin-2; Lymphocyte Activation; Mice; Proto-Oncogene Mas; Proto-Oncogenes; T-Lymphocytes; Transcription, Genetic

The relationship between c-myc expression and thymocyte activation was studied in freshly isolated human thymocytes and in thymocytes activated with various inducing agents. In freshly isolated thymocytes c-myc mRNA is expressed at low levels, while thymocytes activated with Concanavalin A (Con A), the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), Con A in combination with TPA or interleukin 2 (IL-2) are induced to express higher levels of c-myc mRNA. The expression of c-myc is increased within 3 h of stimulation with these inducing agents; the amount of c-myc mRNA which is accumulated is not correlated with the rate of thymocyte proliferation. Dexamethasone and Cyclosporin A (CsA) which inhibit early events of T cell activation and the expression of the interleukin-2 (IL-2) and gamma-interferon (gamma-IFN) genes also markedly suppress the expression of c-myc mRNA in Con A, and Con A + TPA-activated thymocytes. We conclude that activation of c-myc gene expression is an early event observed in activated human thymocytes. The level of c-myc expression is dependent on the mode of thymocyte activation rather than on the rate of thymocyte proliferation. Since freshly isolated thymocytes express low levels of c-myc mRNA it is possible that IL-2 induces c-myc expression at least in a responsive subpopulation of thymocytes during ontogeny.

Topics: Cell Transformation, Neoplastic; Cells, Cultured; Child; Concanavalin A; Cyclosporins; Dexamethasone; Humans; Infant; Lymphocyte Activation; Proto-Oncogenes; RNA, Messenger; T-Lymphocytes; Thymus Gland; Transcription, Genetic

The early passage diploid Syrian hamster embryo (SHE) cells were treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). The treated cells proliferated rapidly; the doubling times shortened; colonies appeared in solid agar medium and transformed foci formed in tissue culture. All of these phenomena suggest that malignant transformation of SHE cell has occurred. Faster cell division rate and multipolar mitosis were demonstrated by time-lapse cinemicrography and scanning electron microscopy. Multipolar mitosis can occur in two forms: direct division and indirect division. The transformed cells were more abundant in microvilli, the number of which increased in accordance with the degree of malignancy. In comparison with the controls, the transformed cells expressed a greater tritiated thymidine incorporation, greater DNA contents and more chromosomes, but no difference in nuclear area. The determination of amino acid changes in media due to the growth of transformed cells showed that the decrease in arginine and increase in ornithine are significant. The results of allogenic animal inoculation suggest that the transformed cells can be characterized into several different stages in the process of transformation.

Topics: Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Chromosome Aberrations; Concanavalin A; Cricetinae; Cyclic AMP; DNA, Neoplasm; Embryo, Mammalian; Mesocricetus; Methylnitronitrosoguanidine; Microscopy, Electron, Scanning; Neoplasm Transplantation

In order to investigate the possibility that the theory of two-stage carcinogenesis can be applied to neurogenic carcinogenesis, we analyzed the promoting effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) on the in vitro malignant transformation of fetal rat brain cells exposed in utero to ethylnitrosourea (ENU). Rat brain cells were transferred to a cultured system at 72 h after a single pulse of ENU (50 mg/kg body weight) to pregnant SD-JCL rats on the 18th day of gestation. The positive findings of glial fibrillary acidic protein and S-100 protein in primary cultured cells by the analysis of immunohistochemistry revealed the neuroglial origin of transformed cells. These cells were divided into 12 groups and were treated twice per week with or without TPA at concentrations from 0.1 to 50.0 ng/ml. From the results of cellular morphology, Concanavalin A agglutinability, colony forming capacity in semisolid soft agar, and tumorigenicity in vivo, malignant transformation of fetal rat brain cells appeared earlier in the ENU group treated with TPA than in the untreated ENU group. On the basis of these observations, it is suggested that TPA might be effective as a tumor promoter on ENU-induced neurogenic carcinogenesis.

Topics: Animals; Brain Neoplasms; Cell Aggregation; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Cocarcinogenesis; Concanavalin A; Ethylnitrosourea; Female; Glial Fibrillary Acidic Protein; Maternal-Fetal Exchange; Neoplasm Transplantation; Neoplastic Stem Cells; Phorbols; Pregnancy; Rats; Tetradecanoylphorbol Acetate

Some major transformation related biological and growth properties have been analysed in an in vitro, system to characterize the PMM-14 cell line, a 20-methylcholanthrene induced transformed mouse embryonic fibroblast cell line. The population doubling time of this cell line was 19 h with moderately high saturation density and plating efficiency. Attachment detachment properties showed reduced adhesion to substratum. Cytogenetic study revealed the existence of a large number of Robertsonian biarmed chromosomes with hypertriploid modal range (60-69) represented by about 21% cells. These cells showed enhanced concanavalin A agglutinability which was reduced by trypsin treatment. Its neoplastic nature was finally established by its ability to grow in agar with a high plating efficiency.

Topics: Agar; Animals; Cell Adhesion; Cell Aggregation; Cell Cycle; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; Fibroblasts; In Vitro Techniques; Karyotyping; Methylcholanthrene; Mice; Trypsin

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Liver Neoplasms, Experimental; Methyldimethylaminoazobenzene; Rabbits; Rats; Surface Properties

Topics: Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Hot Temperature; Humans; Nasopharyngeal Neoplasms; Phenotype

Primary Chinese hamster embryo cultures exposed chronically to 1-methylguanine or 7-methylguanine, modified purines derived from nucleic acid turnover, exhibit a number of properties characteristic of transformed cell lines. One of the earliest effects observed following exposure of cells to the methylated purines is an alteration in cell surface properties as measured by the interaction of the cells with the lectin concanavalin A. Within sixteen hours following inclusion of the compounds in the culture medium, the cells exhibit an increase in concanavalin A mediated hemadsorption. The increase in hemadsorption is accompanied by an alteration in distribution of receptors within the cell population as measured by flow microfluorometry using fluorescin conjugated concanavalin A, and by a decrease in the total number of receptors as measured by binding of radiolabelled concanavalin A. Possible mechanisms for these alterations and their significance for growth control are discussed.

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Cricetulus; Cytoskeleton; Flow Cytometry; Guanine; Hemadsorption; Receptors, Mitogen; Surface Properties; Tritium

Topics: Animals; B-Lymphocytes; Cell Cycle; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cycloheximide; Fibroblasts; Lipopolysaccharides; Lymphocyte Activation; Mice; Oncogenes; Platelet-Derived Growth Factor; RNA, Messenger; Transcription, Genetic

An active supernatant (Ly Con A) was prepared by stimulating the normal human lymph node lymphocytes with Concanavalin A (Con A). Peripheral blood lymphocytes (PBL) from 3 healthy subjects and from 8 patients with chronic lymphocytic leukaemia (CLL) were cultured in the presence of Con A and Ly Con A. The latter induced a significant DNA synthesis both in normal and CLL lymphocytes. A proliferative response was still present in CLL after T lymphocytes depletion. The Con A and Ly Con A treatment also induced morphological changes in CLL lymphocytes consistent with plasma cell differentiation. In 3 cases the appearance of cytoplasmic light chains was detected by immunofluorescence. These findings suggest that peripheral blood B lymphocytes from CLL patients can be stimulated to maturation when helper factor(s) released by mitogen activated lymph node lymphocytes are provided.

Topics: Antibody-Producing Cells; B-Lymphocytes; Cell Transformation, Neoplastic; Concanavalin A; Humans; Immunoglobulin Light Chains; Leukemia, Lymphoid; Lymph Nodes; Lymphocyte Activation; Lymphokines; Thymidine

Purified human peripheral blood T cells that have been depleted of Ia-bearing cells and adherent cells do not proliferate in response to concanavalin A. The addition of as few as 1% radiated L428 tumor cells restores the proliferative capacity of the T cells. The L428 cell line is a long-term tissue culture line of Reed-Sternberg cells obtained from a patient with Hodgkin's disease. The proliferation of the T cells plus the L428 cells follows the same kinetics and has the same response to varying doses of mitogen as either unfractionated mononuclear leukocytes or purified T cells plus allogeneic adherent cells. The L428 cells are 30 times more potent as accessory cells than allogeneic adherent cells. The accessory cell function of the L428 cells is not blocked in cultures containing anti-Ia antibody. Neither supernatant from the L428 cell cultures nor human IL 1 replaces the accessory cells. The ability of the L428 cells to restore the proliferative capacity of purified T cells isolated from patients with active Hodgkin's disease was also studied. Patients with early stages of the disease had normal proliferative responses in the presence of the L428 accessory cells. However, the proliferative response of the poor prognosis, advanced-stage patients was reduced as compared to age- and sex-matched controls, supporting a deficit in their T cell function. The L428 tumor cells share many properties such as accessory cell function, morphology, and cell surface markers with the dendritic cells described in animal and human systems.

Topics: Adult; Antibodies, Monoclonal; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Dose-Response Relationship, Immunologic; Histocompatibility Antigens Class II; Hodgkin Disease; Humans; Interleukin-1; Kinetics; Lymphocyte Activation; Lymphocyte Cooperation; Male; Middle Aged; T-Lymphocytes

Gamma glutamyl transpeptidase (GGT) activity and fluorescein-labeled lectins Concanavalin A (Con A) and Ricinus communis-120 binding sites were studied in lesions produced by 7,12-dimethylbenz[a]anthracene during chemical carcinogenesis using syngeneic heterotopic rat tracheal transplants. Although all types of lesions showed positive GGT activity and lectin binding, prominent differences were observed between the different types of lesions. Compared with regular squamous metaplasia, dysplastic-neoplastic lesions showed much stronger reaction for GGT and lectins; normal and hyperplastic mucociliary epithelium showed intermediate reactions. Although the relationship between keratinization, cell surface changes, GGT activity and lectin binding are not completely clear, lectins seem to be good markers of dysplastic and neoplastic lesions of the respiratory tract epithelium.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Binding Sites; Cell Transformation, Neoplastic; Concanavalin A; Epithelium; Female; gamma-Glutamyltransferase; Lectins; Rats; Rats, Inbred F344; Respiratory Tract Neoplasms

Several Ia+ (BC3A, TA3, D1B) or Ia-inducible (WEHI-3, P388D1) tumor lines were tested for accessory cell function for the activation of antigen-specific T cell proliferation and for the induction of T helper cells that help B cells in antibody production. All lines were able to induce antigen-specific T cell proliferation in an MHC-restricted way, but none activated T helper cells to soluble antigens under all conditions tested. In comparison, starch-induced peritoneal exudate macrophages induced T cell proliferation as well as T cell help. Some of the lines tested induced nonspecific suppressor cells that were Ly-2-positive and partially or completely inhibited antibody responses. The induction of suppressor cells, however, is not the reason for the failure of the tumor lines to activate T helper cells. These data indicate that antigen-specific T cell proliferation and helper activity do not necessarily correlate.

Topics: Animals; Antilymphocyte Serum; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Histocompatibility Antigens Class II; Immune Tolerance; Lymphocyte Activation; Lymphocyte Cooperation; Macrophages; Mice; Mice, Inbred A; Mice, Inbred BALB C; T-Lymphocytes, Helper-Inducer

The DNase I inhibition assay was used for the determination of the relative amounts of monomeric and total actin in normal human peripheral blood lymphocytes, normal Sprague-Dawley rat peritoneal leukocytes, and 6 transformed human cell lines of lymphoid and myeloid origins. In the normal lymphocytes and leukocytes, greater than 50% of the total actin was monomeric. In contrast, only 1 of the 6 transformed lines had greater than 50% of its actin in the monomeric form. In the other 5 lines, the percentage of actin in the monomeric form ranged from 23 to 39%. The normal lymphocytes, peritoneal leukocytes, and 2 of the transformed cell lines (CEM and K562) were examined in greater detail. The total actin (as a percentage of the total protein) was found to be much lower in the transformed cell lines than it was in the normal lymphocytes. The total amount of actin in the normal rat leukocytes was very similar to the amount in the normal human lymphocytes. In addition to these differences between the normal and transformed human cells, treatment of the normal lymphocytes with mitogenic doses of concanavalin A was found to significantly reduce the amount of monomeric actin in the cells. Similar treatment of the transformed cells produced no significant reduction in the monomeric actin.

Topics: Actins; Animals; Cell Transformation, Neoplastic; Concanavalin A; Deoxyribonucleases; Humans; Leukocytes; Lymphocytes; Neutrophils; Rats

The susceptibility of T and B lymphocytes to productive infection and transformation by murine leukemia virus Moloney was determined by enumeration of cells producing infectious virus after in vitro infection of mitogen-stimulated, isolated cell populations and by in vivo infection of euthymic BALB/c and thymus-deficient (nude) mice. Our in vitro results demonstrated that the majority of splenic T cells and thymocytes are resistant to productive infection in vitro; a specific subpopulation of susceptible nylon-adherent splenic T cells was identified, however. Similarly, surface immunoglobulin-positive B cells also represent susceptible targets in vitro; mature B cells, however, did not represent the principal target for transformation in the in vivo experiments. Infected euthymic mice expressed increasing titers of murine leukemia virus and uniformly developed fatal T-cell lymphomas at 10 to 12 weeks postinfection; nude mice, in contrast, maintained high, stable levels of viremia throughout the 28 weeks of observation. Infected nude mice remained free of malignancy or developed either granulocytic leukemias or, in one case, reticulum cell sarcoma. Collectively, the results indicate that while the majority of T cells are resistant to productive infection, they represent the principle targets for transformation; B cells, however, represent permissive targets for virus replication, but are resistant to transformation.

Topics: Animals; B-Lymphocytes; Cell Separation; Cell Transformation, Neoplastic; Cell Transformation, Viral; Cells, Cultured; Concanavalin A; Leukemia, Experimental; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Nude; Moloney murine leukemia virus; T-Lymphocytes; Virus Replication

Human peripheral blood lymphocytes (PBL), obtained from patients with a variety of malignancies, when incubated in vitro with phytohemagglutinin (PHA), lysed fresh autologous tumors during a short-term 51Cr-release assay. These PHA-activated killer (PAK) cells produced maximum lysis of tumor cells by 4 to 8 hr of co-cultivation. PHA incubation induced the generation of cells lytic for autologous and allogeneic tumors but not autologous or allogeneic PBL. Cold target inhibition studies demonstrated that autologous and allogeneic tumors of various histologic types all shared the determinants recognized by PAK cells. Some adherent cells were necessary for generation of these PAK, but higher numbers were suppressive. Augmentation of tumor cell lysis was seen when adherent cells were partially removed before PHA activation. The PAK effector cell was OKT3+, OKT8+. The precursor cell of the PAK was separated from natural killer (NK) cells on Percoll gradients and was generated from thoracic duct lymphocytes, a population devoid of NK cells. Furthermore, the phenotype of the majority of the precursor cells was OKT3+, OKM1-. Activation by PHA for 2 days was found to be an efficient and convenient method for generating lymphoid cells lytic for fresh autologous human tumor. The biologic and possible therapeutic role of these cells is currently being investigated.

Topics: Cell Adhesion; Cell Transformation, Neoplastic; Concanavalin A; Cytotoxicity, Immunologic; Hematopoietic Stem Cells; Humans; Killer Cells, Natural; Kinetics; Lymphocyte Activation; Lymphocytes; Neoplasms; Phenotype; Phytohemagglutinins; Thoracic Duct

A concanavalin A (ConA) derivative of a monovalent monomeric nature, including a monomeric molecular weight at pH 7.4, significantly induces lymphocyte blastoid transformation. The derivative was recently obtained by Tanaka, I., Abe, Y., Hamada, T., Yonemitsu, O., and Ishii, S. ((1981) J. Biochem. (Tokyo) 89, 1643-1646) by a novel procedure of photochemically induced alkylation of tryptophan residues of native tetravalent ConA using a high pressure mercury lamp in the presence of chloroacetamide followed by two steps of column chromatography. This monovalent monomeric ConA (Mm-ConA) was demonstrated to be almost equally potent in producing the maximal response of lymphocytes when compared with native tetrameric ConA, although Mm-ConA was required at about 70 times as high as the concentration of native ConA on a weight basis to attain the maximal response of lymphocyte activation. The binding potency of the former to lymphocytes was about two-thirds as potent as that of the latter. Mm-ConA failed to agglutinate sheep erythrocytes at concentrations 1800-fold higher than native tetravalent ConA, but showed a weak but definite agglutinating activity against guinea pig erythrocytes at a relatively high concentration (approximately 80 micrograms/ml). Cell cluster formation was observed in lymphocyte cultures for 24 to 48 h with Mm-ConA where DNA replication in stimulated lymphocytes was observable. No significant difference was observed between sizes of cell clusters formed in the presence of Mm-ConA and of native tetravalent ConA at this phase of lymphocyte activation. The present results suggest that the multivalency of ConA with respect to sugar-binding sites may not be a stringent requirement for lymphocyte activation, and that another binding site for cell membrane, which has been suggested to exist in the ConA protomer and to be hydrophobic (membranophilic), may play a subsidiary but important role in triggering lymphocyte blastoid transformation as well as hemagglutination with Mm-ConA. The biological significance in lymphocyte activation of lectin valency with respect to sugar-binding sites has been discussed, comparing the effects of various ConA derivatives of different valencies, including a monovalent monomer (Mm-ConA), divalent dimers (beta-ConA, sulfomethylamino-ConA, and succinylated ConA), and tetravalent tetramers (alpha-ConA), on the cell surface.

Topics: Animals; Cell Transformation, Neoplastic; Concanavalin A; Kinetics; Lymphocyte Activation; Lymphocytes; Mice; Mice, Inbred Strains; Spleen

Hybrids between non-malignant mouse 3T3 cells and highly malignant rat JF1 cells were selected in semisolid agar medium without asparagine: 3T3 cells do not grow in agar; JF1 cells required asparagine for their growth. Some surface properties: agglutination by Con A, saturation density, electrophoretic mobility and behaviour in the two-phase aqueous polymer system dextran-polyethylenglycol, -have been studied in the parental cells and in two their hybrids (H9-1 and H9-2). JF1 cells differ from 3T3 cells in their behaviour in two phase polymer system by lower electrophoretic mobility, by higher agglutinability with Con A and higher saturation density. H9-1 and H9-2 hybrids behave in two phase polymer system similar to 3T3 cells, they have low saturation density, intermediate electrophoretic mobility and intermediate agglutinability by Con A. These results indicate that in hybrids between non-malignant 3T3 cells and malignant JF1 cells selected under conditions (semisolid agar) favorite for the growth of oncogenic cells, surface properties of malignant parents are not dominated. Both hybrids are phenotypically similar with drastic genetical difference. By means of C-banding it has been shown that H9-2 hybrid contain approximately one 3T3 and one JF1 genome; H9-1 hybrid contains two JF1 genomes and about 15% of 3T3 genome.

Topics: Animals; Cell Aggregation; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Electrophoresis; Fibroblasts; Gene Expression Regulation; Hybrid Cells; Kinetics; Mice; Rats; Sarcoma, Experimental; Surface Properties

Topics: Animals; B-Lymphocytes; Cell Transformation, Neoplastic; Concanavalin A; Epitopes; Female; Histocompatibility Antigens Class II; Immunoglobulin M; Interleukin-1; Lipopolysaccharides; Lymphokines; Lymphoma; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Proteins; Receptors, Antigen, B-Cell; Time Factors

Topics: Animals; Cell Aggregation; Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Erythroblasts; Erythrocyte Membrane; Erythrocytes; Friend murine leukemia virus; Gelatin; Mice; Microscopy, Electron, Scanning; Microvilli; Muramidase; Ovalbumin; Peptides; Polylysine; Proteins; Sarcosine; Surface Properties

Phytohemagglutinin (PHA) or either of its isolectins, erythroagglutinin or leukoagglutinin, causes a dose-dependent decrease in 12-0-tetradecanoylphorbol 13-acetate (TPA)-promoted transformation of Syrian hamster embryo cells, but has no effect on transformation induced by ultraviolet irradiation. The ineffectiveness of concanavalin A indicates that not every lectin inhibits TPA. Galactose, a dominant sugar in receptors for PHA binding, reverses the inhibition of TPA promotion caused by PHA but galactose does not inhibit TPA promotion itself. Therefore, the TPA and PHA binding sites are functionally discrete. The PHA inhibition of TPA-promoted transformation is reversible because PHA is only effective if present with TPA, whereas lymphotoxin, an immunologic hormone, has a persistent anti-carcinogenic effect, regardless of whether it is added before or after TPA. PHA in conjunction with lymphotoxin causes additional inhibition of TPA-promoted transformation. PHA and lymphotoxin affect the biological activity of TPA by diverse mechanisms. Lymphotoxin alters the physiological state of the cell, causing a change in the cellular response to TPA. PHA may affect either the binding of TPA to a critical cellular receptor for promotion or a later step in promotion.

Topics: Agglutinins; Animals; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Galactose; Leukocytes; Lymphotoxin-alpha; Neoplasms, Radiation-Induced; Phorbols; Phytohemagglutinins; Tetradecanoylphorbol Acetate

From C57BL mouse melanoma B-16 cells, variant clones were selected in vitro which were resistant to the lectins wheat-germ agglutinin and ricin. Cells were also selected which survived toxic concentrations of concanavalin A. Four different in vivo assays using intradermal, intravenous, intraperitoneal and intramuscular injections were used to assess the tumorigenicity and metastasizing capacity of these lectin-resistant variants. It was concluded that to obtain a complete picture of the malignant properties of a given cell line or clone, all four assays have to be carried out. In comparison with the parental cells, the WGA-resistant cells showed a most dramatic decrease in metastasizing capacity through both lymphatic and vascular channels. Tumorigenicity was also reduced. The ricin-resistant cells showed a defective development into lung tumors and thus displayed a reduction in metastasis through the hematogenous route. Since this line did not change its capacity to metastasize via the lymphatic route, and the tumorigenicity was not significantly altered, it will be a good model for studies seeking to dissociate these two properties. The Con-A-selected cells, when injected intravenously, developed tumor nodules in the liver in addition to those in the lungs, while no striking alterations in tumorigenicity or metastasizing capacity could be detected in this line.

Topics: Animals; Cell Line; Cell Survival; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Drug Resistance; Lectins; Melanoma; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental

Topics: Animals; Binding Sites; Binding, Competitive; Cell Transformation, Neoplastic; Concanavalin A; Lymphocyte Activation; Lymphocytes; Macrophage Activation; Macrophages; Mice; Mice, Inbred C57BL; Rats; Rats, Inbred WF; Receptors, Fc

In the previous paper, we reported that ricin and a concanavalin A-ricin A chain conjugate were more toxic to normal cell lines than the transformed variants. In this work, we confirmed this conclusion using other cell lines (KB 3 flat, ST-balb and R4-IudR) which were derived from the Balb/3T3 clone. KN7-8 had a lower number of Con A binding sites than the sensitive parent line (A-31), while the apparent association constant of Con A and surface receptors KN7-8 was slightly higher than that of A-31. On the other hand, both cell lines had almost the same number of ricin binding sites, while the apparent association constant of KN7-8 was lower than that of A-31. These differences in the number of binding sites and the apparent association constant might not be due to the different sensitivity of these cell lines to the toxin, because the amounts of bound 125I-ricin internalized into the two cell lines during incubation at 37 degrees C were essentially the same. Furthermore, no evidence was obtained showing that there was a difference between A-31 and KN7-8 cells in the ability of reduction of the disulfide bond of ricin, degradation of internalized toxin and in the sensitivity of the protein synthesizing machinery.

Topics: Animals; Cell Line; Cell Survival; Cell Transformation, Neoplastic; Concanavalin A; Cytoplasm; Mice; Protein Biosynthesis; Ricin

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

The PFT cell line originated from diploid endometrial cells and was subcultured more than 500 times. Nontumoral, benign and malignant transformation appeared at different stages. Four populations (F1, F2, F3 and F4) of cells were demonstrated sequentially by chromosomes analysis. These consisted of eight phenotypes corresponding to the sequential appearance of markers which included three chromosomal aberrations. The PFT malignant transformation in vitro is a progressive process through qualitatively different stages and may reflect neoplastic development in vivo. It is suggested that the benign and malignant transformations were under separate genetic control since they occurred after two chromosomal translocations on two different chromosomes. The spontaneous expression of a pig endogenous type-C virus which occurred concomitantly with the first translocation appeared to be related to oncogenesis.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; Diploidy; Endometrium; Female; Hydrogen-Ion Concentration; Karyotyping; Mice; Mice, Nude; Microscopy, Electron; Neoplasms, Experimental; Phenotype; Retroviridae; Swine; Translocation, Genetic; Virus Replication

T lymphocytic colony formation by peripheral lymphocytes separated by discontinuous albumin gradient centrifugation was evaluated in 8 patients with Philadelphia (Ph1)-positive chronic myeloid leukemia (CML). Colonies were obtained using a liquid-on-agar culture system recently introduced (PHA overlayer-leukocyte feeder layer assay) which has been shown to be simple and reliable. The pattern of colony growth in CML and in normal controls was similar, the peak ranging from the 4th to the 6th day. Also the morphological aspects of colonies did not differ in the two groups. Cells recovered from CML lymphocytic colonies were shown to belong to T cell lineage, as they are able to form spontaneous E-rosettes and to respond to mitogenic stimulation in vitro. In contrast, cells recovered from all other cultured fractions failed to display these properties. Cytogenetic analysis showed that T colony cells were Ph1-negative whereas the chromosome anomaly was found in nonlymphoid colonies of the same patients, thus suggesting a nonclonal origin of T lymphocytes in CML.

Topics: Agar; Cell Transformation, Neoplastic; Cells, Cultured; Centrifugation, Density Gradient; Chromosomes, Human, 21-22 and Y; Concanavalin A; Humans; Leukemia, Myeloid; Phytohemagglutinins; Pokeweed Mitogens; Rosette Formation; T-Lymphocytes

Monoclonal antibodies recognizing human T cell differentiation antigens were used to study lymphocyte populations in three cutaneous diseases. Neoplastic lymphocytes from patients with varying phases of cutaneous T cell lymphoma (mycosis fungoides, Sezary syndrome and related presentations) were reactive with OKT1 and OKT3 (pan T cell reagents) and OKT4 (an antibody defining the functional "helper" T cell subset). The malignant cells lacked membrane antigens reactive with OKT5 and OKT8 (markers of "suppressor" T cells). The presence of an OKT1+, OKT3+, OKT4+, OKT5-, OKT8- phenotype on the neoplastic T lymphocytes of cutaneous T cell lymphoma (CTCL) supports the clinical impression that all phases of CTCL represent a single disease entity. A patient with pemphigus vulgaris, a disease of autoreactive, antiepidermal antibodies was shown to consistently have a marked expansion of the peripheral blood OKT4 reactive T lymphocyte population. These findings suggest that autoantibodies in pemphigus vulgaris may occur in the context of a profound OKT4/OKT5 immunoregulatory imbalance. Peripheral blood lymphocytes from patients ith extensive psoriasis vulgaris had a normal profile of reactivity with the OKT antibodies. In addition, OKT6 (marker of intrathymic T cells) has been shown to react with Ia+ dendritic cells in the epidermis suggesting that this antibody may recognize Langerhans' cells.

Topics: Adolescent; Adult; Aged; Animals; Antibodies, Monoclonal; Cell Transformation, Neoplastic; Concanavalin A; Humans; Lymph Nodes; Lymphocytes; Lymphoma; Mice; Middle Aged; Pemphigus; Phenotype; Psoriasis; Rabbits; Skin Diseases; T-Lymphocytes

We have studied the expression of transformation-related parameters in tertiary hybrid clones from an apparently transformed human cell strain (ME) and a malignant mouse cell line (A9). The derivation of these hybrid clones, chromosome analysis, and tumorigenic potential have been reported elsewhere (1). In the present work, the serum sensitivity, saturation density, cloning efficiency on plastic surfaces and in semisolid medium, survival in aggregate form, concanavalin A agglutinability, sugar uptake, and secretion of plasminogen activator in 4 tertiary hybrid clones and in both parent cells have been determined. While, in general, the extent to which these transformation-related traits were expressed in the hybrid clones closely resembled the malignant A9 parent cell, all hybrids displayed partial to complete suppression of tumorigenicity. These results suggest no apparent relationship between the extent of quality of transformation and tumorigenicity in these cell hybrids.

Topics: Agglutination; Animals; Blood; Cell Adhesion; Cell Aggregation; Cell Division; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Deoxyglucose; Humans; Hybrid Cells; Mice; Neoplasms, Experimental; Plasminogen Activators

Cells of a pig Fallopian tube line were agglutinated by concanavalin A (Con A). Similar ConA agglutination was shown in diploid and nontumoral transformed cells with one (T1) translocation until the 112th subculture. Then a progressive increase of agglutination was shown until the 149th subculture and the rate of agglutination remained constant thereafter. The progression of ConA agglutination paralleled the progression of anchorage independence. The highest ConA agglutination occurred when anchorage independence was well established and persisted until the 193rd T1 subculture when benign tumorigenicity was demonstrated. After the appearance of the second translocation (T2), the self-agglutination of the cells was directly related to malignancy and to high tumor incidence in mice. Anchorage independence and the highest ConA agglutination were directly related to benign transformation and indirectly related to malignant transformation of the pig Fallopian tube cell line.

Topics: Agglutination; Animals; Cell Adhesion; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Fallopian Tubes; Female; Methylglucosides; Swine

The effect of anaplastic carcinoma (15091A) and fibroblast cell culture supernates was examined in conjunction with Concanavalin A (Con A) on the splenic lymphocyte transformation of syngeneic A/J mice. Though both the tumor and normal cell culture supernates caused a dose-dependent suppression of the in vitro DNA synthesis of lymphocytes, larger depressive effects were observed with the former. Similar results were observed with supernates collected from tumor cell and fibroblast cultures in 10% heat inactivated fetal calf serum or serum-free media. The results of these experiments indicate that the depressed immunological reactivity observed in animals and human with cancer may be attributed to enhanced release of immunosuppressive factor(s) by malignant tissue into the body fluids of hosts.

Topics: Animals; Carcinoma; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; DNA; Fibroblasts; Immunosuppression Therapy; Immunosuppressive Agents; Male; Mice; Mice, Inbred A

Differentially L-fucose-labelled glycopeptides from the surface of a Syrian golden hamster (SGH) fetal lung control cell line were compared with those from chemically-transformed and tumour cell lines derived from the control line by cochromatography on Concanavalin A-Sepharose (Con A-Sepharose) and Sephadex G-50. Quantitative differences were found both in the unbound and specifically-bound fractions between control and transformed cells upon Con A-Sepharose chromatography. In the glycopeptides from transformed and tumour cells, the unretarded fraction was concomitantly decreased compared to the controls. When the ratio of unbound to specifically-bound fractions was used, a statistically significant difference could be calculated between the values of control versus transformed or tumour cells. In all transformed and tumour cell lines investigated, the quantitative change in Concanavalin A binding, expressed as an increase of the ratio of unretarded to specifically-bound glycopeptides, was paralleled by a shift of transformed or tumour glycopeptides to higher apparent molecular weight compared to the control in gel chromatography.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fetus; Glycopeptides; Lung; Membrane Proteins; Mesocricetus; Neoplasms, Experimental; Sepharose

Topics: Animals; Cell Communication; Cell Transformation, Neoplastic; Concanavalin A; Lymphocytes; Neutrophils; Rats; Rats, Inbred Strains; Sarcoma, Experimental

Sensitivities of human transformed cell line RSa and its variant cell line IFr to the cytotoxicity of Con A were compared. IFr cells were more resistant than RSa to Con A. Con A-resistant cell lines, Con Ar-1 and Con Ar-3, were isolated from RSa, and they were slightly more sensitive than RSa cells to the cell growth-inhibitory actions of interferons. Agglutinability of RSa, IFr, and Con Ar cells by Con A was compared and found to be almost equal. The combined effects of Con A and interferon upon growth and viability of these cell lines were tested. When RSa and IFr cells were treated simultaneously with Con A and Le-IF, growth of the cells was suppressed more markedly than when treatment was with Con A or Le-IF alone. To clarify the mechanism of this phenomenon, binding of 125I-labeled Con A was examined. Though ther wee some differences, both leukocyte and fibroblast interferon enhanced the binding of Con A to RSa cells and also in Con Ar cells but, in interferon-resistant IFr and HEC cells, enhancement of Con A binding was low or not observed. Therefore, the combined effect of Con A and interferon on the inhibition of cell growth is not considered to be merely due to the enhanced binding of Con A by interferon action. Successive treatment of RSa or Con Ar cells with Con A and interferon did not enhance the antiviral action of interferon at all. On the contrary, simultaneous treatment with Con A and interferon suppressed the antiviral action of interferon, depending on the concentration of Con A used. Thus, the effect of Con A on the antiviral and cell growth-inhibitory actin of interferon seems rather different.

Topics: Cell Aggregation; Cell Division; Cell Survival; Cell Transformation, Neoplastic; Concanavalin A; Drug Resistance; Humans; Interferons; Vesicular stomatitis Indiana virus; Virus Replication

Suppressor cells in spleens of mice bearing the Lewis lung carcinoma (3LL) suppressed the anti-tumor immune response in vivo but not in vitro. The hydrocortisone (HC)-resistant fraction of spleen cells from tumor-bearing mice (TBM) was, on the other hand, suppressive in vitro, due to enrichment for HC-resistant suppressor cells. This cellular fraction suppressed the tumor-induced differentiation of memory cells as well as the tumor-independent in vitro activation of TBM spleen cells. The in vitro lymphoproliferative response of TBM spleen cells to mitogens was also subjected to suppression: these cells showed a lower response to a mitogen such as Concanavalin A (Con A) and were capable of suppressing the response of normal spleen cells to this mitogen. HC treatment of TBM 2 days prior to spleen harvest enriched the splenic population for suppressor cells that suppressed Con A-induced activation.

Topics: Animals; Cell Transformation, Neoplastic; Concanavalin A; Cytotoxicity, Immunologic; Drug Resistance; Hydrocortisone; Lung Neoplasms; Male; Mice; Neoplasms, Experimental; Spleen; T-Lymphocytes; T-Lymphocytes, Regulatory

The lecture reviews some aspects of the work on the analysis of malignancy that have been, and are now being, pursured in the Dunn School. A brief outline of the early experiments that first demonstrated that the malignancy of mouse tumor cells can be suppressed by the fusion with normal cells is given, and then two areas of current interest in the laboratory are described. The first is an attempt to analyze the clinically important property of tumors to metastasize and the second is the work on the isolation and identification of an abnormal membrane glycoprotein present in tumor cells. In addition the value of cell fusion methods as a general test of hypotheses of malignancy is emphasized.

Topics: Animals; Autoradiography; Cell Fusion; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Glucose; Glycoproteins; Humans; Hybrid Cells; Lectins; Membrane Proteins; Mice; Neoplasm Metastasis; Neoplasm Proteins; Neoplasms, Experimental

Topics: Adsorption; Cell Cycle; Cell Line; Cell Membrane; Cell Survival; Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Erythrocytes; Fibroblasts; Humans

Neoplastic transformation of primary fibroblast culture derived from esophagus tissue of a 52-year old male esophageal cancer patient was induced by chemical carcinogen (N-methyl-N'-nitro-N-nitrosoguanidine, MNNG) treatment. The transformed cells showed the biological and morphological properties characteristic of malignant cells, such as loss of contact inhibition, unlimited growth in vitro, aneuploidy, agglutinability by concanavalin A, formation of microvilli on the cell surface, growth on solid agar medium and tumor (fibrosarcoma) formation after heterotransplantation into immunosuppressed newborn mice.

Topics: Aneuploidy; Animals; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Disease Susceptibility; Esophageal Neoplasms; Esophagus; Fibroblasts; Fibrosarcoma; Humans; Male; Methylnitronitrosoguanidine; Mice; Middle Aged; Neoplasm Transplantation

Topics: Animals; Cell Adhesion; Cell Line; Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Dogs; Epithelium; Fibroblasts; Humans; Mice; Statistics as Topic

A method for the in vitro identification of transformed rat liver epithelial and hepatoma cells was developed using the preferential microagglutination of concanavalin A (Con A) coupled to agarose beads (Con A:agarose) to their colonies. Con A:agarose attaches to the cell surface through a specific interaction of the Con A moiety and its receptors. The attachment is dependent on the mobility and aggregation of the Con A: receptor complex on the membrane. Agents which interfere with the interaction reduced the bead density over the colonies. For the quantitative determination of transformed colonies in a mixed-cell population, also containing untransformed cells, it is essential to compare colonies of a similar size or to use the bead density per unit area as the index. When a variety of rat liver epithelial cell lines were tested, the assay proved to be simple, reproducible, and precise. It was found that the increased attachment of Con A:agarose to cell colonies is a characteristic of transformed or malignant rat liver cells.

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Liver; Liver Neoplasms, Experimental; Liver Regeneration; Rats; Sepharose

Topics: 9,10-Dimethyl-1,2-benzanthracene; Adenoviruses, Human; Animals; Benzopyrenes; Cell Aggregation; Cell Division; Cell Transformation, Neoplastic; Cell Transformation, Viral; Clone Cells; Cocarcinogenesis; Concanavalin A; Embryo, Mammalian; Membrane Proteins; Phenotype; Phorbols; Plasminogen Activators; Rats; Tetradecanoylphorbol Acetate

Concanavalin A (Con A)-mediated red blood cell (RBC) adsorption with the RBC coating method (in which Con A-coated RBC's are adsorbed to fibroblasts) was greatly increased by glutaraldehyde fixation of RBCs before Con A-coating and decreased by the fixation of fibroblasts. On the other hand, RBC adsorption with the fibroblast coating method (in which RBCs are adsorbed to Con A-coated fibroblasts) was decreased by the fixation of RBCs and increased by the fixation of fibroblasts before Con A coating. The fixation of RBCs or fibroblasts after Con A coating did not have these effects. In addition, the fixation of both RBCs and fibroblasts nearly completely abolished RBC adsorption with either method. However, the amount of [3H] Con A binding was not affected by the fixation. RBC adsorption with the fibroblast coating method was also affected by cytochalasin B, colchicine, NaN3 and dibucane treatments of fibroblasts. These drug treatments of fibroblasts, however, did not affect RBC adsorption with the RBC coating method, except cytochalasin B. In addition, the effects of drug treatments of fibroblasts examined occurred nearly to the same extent for nonsenescent, senescent, and transformed cells. Our results suggest that secondary processes after Con A binding, receptor mobility and receptor association with cytoskeletals, play important roles in RBC adsorption, but that the roles do not change with aging or transformation.

Topics: Aldehydes; Azides; Calcimycin; Cell Survival; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Cytochalasin B; Dibucaine; Erythrocytes; Female; Fibroblasts; Fixatives; Glutaral; Hemadsorption; Humans

Answers are beginning to emerge to the questions posed in the introduction to the preceding section. In vitro techniques that allow characterization of malignant cells have particular relevance when, as in Hodgkin's disease, the precise identity of the cells remains in doubt. Monolayer tissue cultures derived from Hodgkin's disease tumours and maintained as established cell lines have proven amenable to a variety of cytogenetic, immunological, enzymatic, and ultrastructural studies. Tissue culture experiemnts, in conjunction with meticulous immunological studies of individual Reed-Sternberg cells from non-cultured tumours, suggest that neoplastic cells of Hodgkin's disease are related to, and possibly derived from, cells of the monocyte-macrophage system. The lymphocytes that comprise an integral part of the cellular proliferation and form the basis for histological subclassification of the tumour could be a manifestation of cell-mediated immunity against this non-lymphoid malignant cell. The immunodeficiency of patients with untreated Hodgkin's disease of limited anatomical extent is not the primary event of the disorder and probably not related to the site at which the aetiological agent acts. The deficit does not result solely from impaired T-cell function and appears to arise as a consequence of excessive suppressor cell activity. Inhibitory monocyte-lymphocyte interactions may be one of the causes of defective cell-mediated immunity in Hodgkin's disease. The possible significance of elevated levels of circulating immune complexes in the serum of patients with Hodgkin's disease is indicated by the finding that such complexes react with cells of long-term monolayer tissue cultures derived from the tumour. Circulating immune complexes may be one source for intracellular immunoglobulin in non-cultured Hodgkin's disease cells. The presence of polyclonal immunoglobulin G on the membrane and within the cytoplasm of Reed-Sternberg cells could be due to in vivo binding and ingestion of immune complexes by such cells. The specificity of the interaction between soluble complement-containing immune complexes and neoplastic cells of Hodgkin's disease depends on the nature of the complexed antigen. The complexes could non-specifically attach via an Fc receptor or, if the complexed antigen is identical to a tumour cell antigen, the binding could be specific. If the immune complexes are tumour specific they could provide a source for isolation and identification of

Topics: Antigen-Antibody Complex; Antigens, Neoplasm; Binding Sites; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Hodgkin Disease; Humans; Immunologic Deficiency Syndromes; Kinetics; Retroviridae; T-Lymphocytes, Regulatory

The effect of thymosin on suppressor-cell function was evaluated in vivo in a murine tumor system and in vitro on human lymphocytes. In mice, the Lewis tumor system was used. We showed that splenocytes from tumor-bearing animals were able to enhance tumor growth in a syngeneic system. This enhancement was similar when thymocytes from tumor-bearing animals were used and disappeared after anti-Thy 1-2 antiserum treatment, suggesting a T-dependence. Treatment of the tumor-growth-enhancing lymphocytes with corticosteroids or irradiation caused this effect to disappear completely suggesting that the tumor-growth-enhancing T-lymphocytes were suppressor T-cells. Furthermore thymosin (fraction 5)-treated, tumor-growth-enhancing T-lymphocytes were not able to enhance tumor growth and even significantly decreased it. In the human system we showed that Con A-stimulated lymphocytes were able to suppress the response of normal lymphocytes to PHA, PWM, and Con A, and in MLC. This effect was significantly blocked in presence of thymosin fraction 5.

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Humans; Immune Sera; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred C57BL; Neoplasms, Experimental; Phytohemagglutinins; Spleen; T-Lymphocytes; Thymosin; Thymus Hormones

Topics: Animals; Cell Count; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Female; Fibrosarcoma; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Male; Mitogens; Simian virus 40; Spleen; Thymus Gland

Topics: Burkitt Lymphoma; Cell Line; Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Herpesvirus 4, Human; Humans; Infectious Mononucleosis; Lectins; Lymphocyte Activation; Lymphocytes; Protein Biosynthesis

Topics: B-Lymphocytes; Burkitt Lymphoma; Cell Differentiation; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Herpesvirus 4, Human; Humans; Lymphocyte Activation; Membrane Proteins; Molecular Weight; Neoplasm Proteins; T-Lymphocytes

The antiviral and anticellular activity of partially purified mouse interferon has been tested in cell lines transformed with Simian Virus 40 (MEB-SV 40), mouse sarcoma virus (Harvey strain) (MEB-MSV), and 20-methylcholanthrene (MEB-MCH), respectively. The transformed lines were derived from C3H mouse embryonic primary cells. It has been shown that the MEB-MSV cells were 10 to 50 times less sensitive to the antiviral effect of interferon than the MEB-SV 40 or MEB-MCH cells. A 30% reduction of the number of treated cells as compared with untreated control cells was taken as basis for comparison of anticellular activity of interferon in transformed lines. While the MEB-MCH cells required 1000 units of interferon for a 30% growth inhibition, about 3000 units were necessary for a comparable suppression of MEB-SV 40 cells and/or MEB-MSV cells.

Topics: Agglutination; Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Embryo, Mammalian; Interferons; Methylcholanthrene; Mice; Mice, Inbred C3H; Newcastle disease virus; Sarcoma Viruses, Murine; Simian virus 40

Topics: Acid Phosphatase; Animals; Cell Transformation, Neoplastic; Concanavalin A; Macrophages; Mice; Mycobacterium bovis; Nucleotidases; Phagocytosis; Propionibacterium acnes; Receptors, Complement; Saccharomyces cerevisiae; Thioglycolates

For the development of therapeutic agents that possess tissue-specific carriers, a method was devised to synthesize an artificial protein hybrid conjugate containing a moiety which binds to a cell membrane receptor and an active fragment of a toxic protein. By the introduction of an activated sulfhydryl group into concanavalin A (Con A), a conjugate of Con A and the ricin A-chain cross-linked with a disulfide linkage was synthesized. The purified conjugate was studied with regard to its inhibitory activity against protein synthesis in cell-free and cultured cell systems. The Con A-rich A-chain conjugate retained about one-third the inhibitory activity of ricin in a cell-free protein synthesis system. It also was highly toxic to cultured normal cells. These results indicate that the conjugate is a structural and functional analog of ricin and that the original membrane-binding chain (B-chain of ricin) could be replaced by Con A. Transformed cells were insensitive to this conjugate and required a longer preincubation time. The sensitivity of the normal cells was reduced in the presence of local anesthetics.

Topics: Animals; Antineoplastic Agents; Cell Survival; Cell Transformation, Neoplastic; Cell-Free System; Cells, Cultured; Concanavalin A; Humans; Protein Biosynthesis; Ricin

Stimulation with bacterial lipopolysaccharide (LPS) of splenic B-lymphocytes infected in vitro with Friend virus complex increased the number of cells with replicating murine leukemia virus (MuLV) [i.e., infectious centers (IC)] up to 100-fold. Concanavalin A (Con A) did not have such an effect. However, the addition of Con A to the LPS-stimulated cultures decreased the number of IC. The inhibitory concentration of Con A (2.5 microgram/ml) was eightfold less than that capable of neutralizing the in vitro infectivity of MuLV (20 microgram/ml). The effect of Con A was not mediated by T-cells; the inhibition of infection was comparable with use of whole spleen cell suspensions from normal BALB/c mice, with T-cell-depleted cell suspensions, or with spleen cells with congenitally athymic nude mice. However, specific removal of Con A from the surface of B-cells with alpha-methyl-D-mannopyranoside prior to the infection reversed the inhibitory effect entirely. It is suggested that the lectin interferes with MuLV on the membrane of B-cells.

Topics: Animals; B-Lymphocytes; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Female; Friend murine leukemia virus; In Vitro Techniques; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; Spleen; Virus Replication

N-Butyl-N-(4-hydroxybutyl)nitrosamine was given to male Wistar rats at a dose of 0.05% in the drinking water for one to five weeks, and agglutination of cell isolated from their bladder by concanavalin A (Con A) was determined at intervals during and after treatment. Mucosal cells were isolated from everted bladder by ethylenediaminetetraacetate treatment and sonication. As early as one week after the start of treatment, Con A caused some agglutination of isolated bladder cells, and this agglutination increased with time, reaching an almost constant value from the third week. Con A agglutination of bladder cells induced by N-butyl-N-(4-hydroxybutyl)nitrosamine treatment for only one week appeared to be irreversible, and it was still observed two weeks after the end of treatment. Scanning electron microscopy showed that microvilli developed on the luminal surface of mucosal cells in situ at the time when the isolated cells became agglutinable with Con A. Measurement of agglutinability of isolated bladder cells with Con A might be a useful way of detecting very early changes in bladder carcinogenesis.

Topics: Agglutination; Animals; Butylhydroxybutylnitrosamine; Cell Transformation, Neoplastic; Concanavalin A; Male; Microscopy, Electron; Mucous Membrane; Nitrosamines; Rats; Time Factors; Urinary Bladder

Topics: Animals; Arginine; Ascitic Fluid; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Culture Media; Cytotoxicity, Immunologic; Macrophages; Male; Mice; Propionibacterium acnes; Thioglycolates

The effect of treatment with trypsin and neuraminidase on morphology, proliferation pattern, adhesiveness and ultrastructure of human normal mammary and carcinoma cell lines has been investigated. These criteria were chosen because they reflect on the cell surface function. Active enzymes were required to produce changes. If trypsinized and then allowed to grow in culture media supplemented with 'fibroblast growth-promoting factor', human normal mammary epithelial cells proliferated in a disorganized pattern with cells overlapping and piling up and developing ultrastructural characteristics of neoplastic cells. On the other hand, if treated with neuraminidase and then permitted to grow in presence of the 'fibroblast growth-promoting factor', mammary carcinoma cells proliferated in an organized pattern, forming one-cell-thick, fibroblast-like monolayers and ultrastructural characteristics of normal epithelial cells.

Topics: Breast; Breast Neoplasms; Cell Adhesion; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Deoxyglucose; Epithelial Cells; Female; Growth Substances; Humans; Neuraminidase; Trypsin

The reverse transformation reaction of Chinese hamster ovary cells from compact, epithelial-like, randomly growing, heavily knobbed, lectin reactive cells into stretched, tighly adherent, smooth-surfaced, lectin resistant, fibroblast-like cells normally elicited by dibutyryl cAMP can be produced to its complete extent by N6-monobutyryl cAMP or 8-bromo-cAMP, O2'-monobutyryl cAMP is ineffective as is cAMP itself in the absence of an inhibitor of phosphodiesterase activity. In the presence of a phosphodiesterase inhibitor, cAMP is fully effective. These results indicate that the role of the butyryl groups of dibutyryl cAMP and, especially, the N6-butyryl, in the reverse transformation reaction is protection of the cAMP analogue from degradation. Butyrate at concentrations of about 1 mM does produce a response which to some extent mimics that of cAMP analogues. The cells, however, fail to assume a fibroblastic-like shape, but rather become flattened. The butyrate effect is much slower and less readily reversible than that evoked by cAMP analogues. Butyrate produces an approximately 2-fold increase in intracellular cAMP levels. These results are consistent with the hypothesis that butyrate effects, in part, are mediated by AMP.

Topics: Butyrates; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cyclic AMP; Testolactone; Theophylline

Succinylated concanavalin A reversibly inhibits the growth of SV40 transformed mouse 3T3 cells and thus causes an accumulation of the cells in the G1 phase of the cell cycle. In a soft substrate (methylcellulose) succinylated concanavalin A also restores in transformed cells the growth behavior typical of untransformed cells.

Topics: Cell Adhesion; Cell Cycle; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Simian virus 40; Succinates

Human cell hybrids derived from malignant HeLa and normal fibroblast parental cells expressed many of the transformed properties of the HeLa parent but their tumor-producing capability was suppressed. Hybrids derived from HeLa/HeLa fusions retained both their transformed and malignant phenotypes. Thus, an apparent separation of the control of the transformed versus malignant phenotype is indicated. Furthermore, several transformed properties--including lack of density-dependent inhibition of growth, lectin agglutination, lowered requirement for serum growth factors, and anchorage independence--are expressed coordinately in the nontumorigenic hybrids. This finding suggests that none of these properties by themselves, or in concert, endows a cell with tumorigenic potential.

Topics: Animals; Blood; Cell Aggregation; Cell Division; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition; Culture Media; Growth Substances; Humans; Hybrid Cells; Mice; Neoplasms, Experimental

Topics: Animals; Anura; Cathepsins; Cell Adhesion; Cell Aggregation; Cell Division; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; DNA, Neoplasm; Epithelium; In Vitro Techniques; Leupeptins; Male; Nitrosamines; Oligopeptides; Peptide Hydrolases; Rabbits; Rana catesbeiana; Urinary Bladder

When cultured in-vitro, originating from different breast cancer patients, tumor cells, identified histologically as carcinoma cells, varied in their proliferation patterns and cell morphology. If exposed for brief periods to vibrio cholera neuraminidaes (VCN), the amount of sialic acid released from the cells varied from one culture to another and increased with higher enzyme concentrations. If exposed to trypsin, the amount of released proteins varied also from one culture to another. Significant difference was observed between the effect of VCN or collagenase on normal and neoplastic cell cultures. Whether human or murine cell cultures, the cell-free media harvested from cultures of neoplastic cells containing high concentrations of collagenolytic-caseinolytic-fibrinolytic and esterolytic activities. Two effects of concanavalin A (Con A) have been distinguished on thymidine incorporation, the first is a decrease in the maximal thymidine uptake, whereas the second is a shift to the maximum thymidine uptake to higher Con A concentrations. At low concentrations, alpha-1-antitrypsin (AAT) had no effect, but at high concentrations it inhibited 3H-thymidine uptake. At low concentrations human profibrinolysin inhibited and at higher concentration sit enhanced uptake of the labeled precursor. Therefore, the collagen olytic caseinolytic-fibrinolytic enzyme is a pacemaker for proliferation of human mammary carcinoma cells.

Topics: alpha 1-Antitrypsin; Breast Neoplasms; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Enzyme Activation; Humans; Mitosis; Plasminogen

Topics: Animals; B-Lymphocytes; Cell Transformation, Neoplastic; Concanavalin A; Fluorescent Antibody Technique; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Neoplasms, Experimental; Pyrans; Radiation Chimera; T-Lymphocytes

Rat embryo cells of low passage subjected to a single treatment with certain carcinogenic fluorenylhydroxamic acids and their respective acetate esters showed signs of transformation in vitro, such as changes in phenotype, growth in soft agar and agglutination with concanavalin A. In addition, certain changes in karyotype and loss of diploidy were observed. There was no evidence, either by electron microscopy or by assay of RNA-dependent DNA polymerase, for the presence of virus. None of these cell lines produced tumors after inoculation into the isologous host. The results of these study lead us to suggest that malignant transformation is a multistep process and that certain criteria of transformation of rat embryo cells are associated with the initial stage(s) in which the cells are transformed without being tumorigenic. The ultimate test for malignant transformation of rat embryo cells remains the production of tumors in a susceptible host after inoculation of treated cells.

Topics: Agglutination Tests; Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Chromosomes; Concanavalin A; Esters; Fluorenes; Hydroxyacetylaminofluorene; Neoplasm Transplantation; Rats

Analysis of six different cell types of normal and transformed fibroblasts grown in vitro and of four different cell types of normal and leukemic lymphocytes grown in vivo have shown a marked decrease of 3- to 30-fold in the specific activity of 5'-nucleotidase in the malignant cells as compared to their normal parental cells. The results have also indicated that a serum stimulation of untransformed or normal fibroblasts and a stimulation of normal lymphocytes by concanavalin A resulted in a significant decrease in the specific activity of 5'-nucleotidase of the stimulated cultures as compared to the resting cells. In both the malignant cells and the stimulated normal cells, the decrease in 5'-nucleotidase activity was not accompanied by a similar decrease in the specific activity of acid phosphatase, indicating a specific enzyme alteration in the surface membranes of the transformed and the normal stimulated cells.

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Culture Media; Fibroblasts; Leukemia, Experimental; Lymphocyte Activation; Lymphocytes; Mice; Mice, Inbred A; Mice, Inbred C57BL; Nucleotidases

The transformed properties of five hybrid cell lines which had either one or another parent in common were studied and compared with their tumorigenicity. Three hybrid cell lines, derived from the Chinese hamster DC-3F/ADX/Aza line, were resistant to actinomycin-D. This property seemed to be correlated with the presence of a marker chromosome from the common parent. The tumorigenicity was intermediate between those of the parent cell lines. On the other hand, agglutinability by concanavalin A (Con A) was variable. Three hybrid cell lines which had either the A9 or the clone 1D (both derived from mouse fibroblasts) showed very similar transformed characteristics, but two were tumorigenic and one not so. It appears from this study that the properties of the hybrid cell lines can be influenced more by one parent, depending on the genes retained at chromosome segregation. The limits of Con A agglutination as a characteristic of transformation and the validity of the check pouch grafts as tumorigenicity test for malignant human cell lines are discussed.

Topics: Agglutination; Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Culture Media; Dactinomycin; Drug Resistance; Genes; Humans; Hybrid Cells; Mice; Neoplasm Transplantation; Neoplasms, Experimental

The comparative aspects of cell growth, i.e., [3H]thymidine and [14C]leucine uptake of low-cancer (P4Bis) and high-cancer (P4BisT) cell lines and of their normal counterparts, have been studied in the presence and absence of concanavalin and Robinia lectins. These lectins have similar effects on cell growth, on thymidine and leucine uptake, and on incorporation of these precursors. The growth of normal cells is stimulated by both lectins, whereas the growth of transformed cells is inhibited. In all cases the uptake of both leucine and thymidine by cells is increased by the lectins, but the percentage of incorporation of the precursors is affected in a different manner. The percentage of thymidine incorporated by normal and transformed cells increases or decreases in direct proportion to cell growth; leucine incorporation is not affected significantly. The reversibility of these lectin effects by specific inhibitors shows that cell membranes are implicated in these phenomena. Our study with normal and transformed cells suggests that cell surface may play a role in the process of malignant transformation and that P4Bis cells are "transitory" between PB1 normal cells and P4BisT high-cancer cells.

Topics: Animals; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Lectins; Leucine; Lung; Lung Neoplasms; Mice; Neoplasm Transplantation; Neoplasms, Experimental; Thymidine; Time Factors; Transplantation, Isogeneic

A malignant cell line derived from the s.c. inoculation of Adenovirus 12 into a CBA mouse has been isolated in vitro, cloned, and within 10 passages the clones have been investigated for their karyotype, morphology, growth rate, saturation density and response to plant lectin in vitro, and their tumorigenicity and growth rate in vivo. The cell lines rapidly acquired a highly heterogeneous karyotype, but remained homogeneous with respect to more complex physiological parameters. Examination of the cellular characteristics has indicated that the rate of growth of the cell lines in vivo, but not their tumorigenicity, may be related to their in vitro potentials. The clones responded differently to the cytotoxic effects of concanavalin A, but there was no correlation between the effect of the lectin and the malignant potential of the cells.

Topics: Animals; Cell Division; Cell Line; Cell Survival; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Karyotyping; Mice; Mice, Inbred CBA; Neoplasm Transplantation; Neoplasms, Experimental; Transplantation, Homologous

Sixty-two explants from peripheral blood, bone marrow and cerebral fluid of children with acute lymphoblastic leukemia (ALL) and leukemic transformed non-Hodgkin lymphoma (NHL) were cultivated for at least 8 weeks. Although lymphatic cells persisted up to 16 weeks in tissue culture, no proliferation was observed in 54 cultures. From the remaining cultures, eight permanently growing cell lines were obtained. Five of these were EBNA (Epstein-Barr virus-specific nuclear antigen)-positive. Three, however, were ENBA-negative and lacked Epstein-Barr virus genomes. Two cell lines (KM-3 and SH-2) expressed neither B nor T cell characteristics. One line (JM) expressed T cell characteristics and complement receptors. The growing lymphatic cells represented leukemic cells, since the pattern of cytochemical staining and that of membrane receptors of lymphoblasts from the same donor prior to cultivation were identical. All leukemic cell lines were derived from patients in relapse. In contrast, no proliferation of leukemic cells occurred in explains from patients revealing the first manifestation of the disease. These results suggest enhanced growth potential of lymphoblasts resisting antileukemic therapy.

Topics: Adolescent; Antigens, Viral; beta 2-Microglobulin; Binding Sites; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Child; Complement System Proteins; Concanavalin A; DNA, Viral; Fluorescent Antibody Technique; Herpesvirus 4, Human; Humans; Leukemia, Lymphoid; Lymphocytes; Lymphoma; Male; Microscopy, Electron; Receptors, Antigen, B-Cell; Staining and Labeling; T-Lymphocytes

Topics: Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Epitopes; Lymphocyte Activation; T-Lymphocytes

The net rate of spontaneous aggregation of cells suspended with EDTA was measured for various cell types including spontaneous transformants and cells transformed with DNA and RNA viruses. The anchorage dependence as determined by growth in methyl cellulose and the tumorigenicity in vivo were also determined. All cells that had lost their anchorage dependency and were tumorigenic showed a high net rate of spontaneous adhesion. A31 was the only nontransformed cell line to have a high net rate of adhesion. The net rate of spontaneous aggregation of cells is a quick and reliable index of tumorigenicity and offers a new approach to understanding the mechanisms of cell surface changes associated with transformation.

Topics: Cell Adhesion; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Edetic Acid; Methylcellulose; Neoplasms, Experimental; Polyomavirus; Sarcoma Viruses, Murine; Simian virus 40

Eight transformation-defective, temperature-sensitive (ts) mutants of the Prague strain of Rous sarcoma virus, subgroup A, have been isolated after mutagenesis with 5-bromodeoxyuridine followed by selection on the basis of focus tests. Five of these mutants, ts GI201, GI202, GI203, GI204, and GI205, exhibit properties like most previously reported isolates in that they show a temperature-sensitive response to each of a variety of transformation-specific parameters tested. Interestingly, GI201, in addition to the temperature-sensitive defect, carries a lesion that was observed as a nonconditional loss of expression of plasminogen activator protease. Three mutants, ts GI251, GI252, and GI253 have been disignated partial transformation-defective (PTD) mutants since they behave as ts mutants according to some tests for transformation and as wild type according to others. These three mutants fail to form foci at the nonpermissive temperature (41 degrees C) and art nontumorigenic in 3-week-old chickens (body temperature, 42 degrees C). The agglutinability by concanavalin A of cells infected with these mutants shows a definite temperature sensitivity, as do the rate of 2-deoxyglucose uptake and the disappearance of the 250, 000-dalton normal cell glycoprotein (large, external, transformation sensitive [LETS]). Although the PTD mutant-infected cells, unlike cells infected with other transformation mutants, exhibit a cell-bound plasminogen activator protease at the nonpermissive temperature, this activator is not detectable as a free protease in the medium, as it is with wild-type, virus-infected cells. The PTD mutants behave like the wild-type parent in their ability to induce transformed growth properties in the infected cells, i.e., growth beyond normal cell saturation density with or without serum-supplemented medium and growth leading to colony formation in soft-agar- or methyl cellulose-containing suspension media.

Topics: Agglutination; Avian Sarcoma Viruses; Bromodeoxyuridine; Cell Transformation, Neoplastic; Concanavalin A; Culture Techniques; Deoxyglucose; Glycoproteins; Mutagens; Mutation; Phenotype; Plasminogen Activators; Temperature

Cellular microtubules, microfilaments, and surface receptors have been postulated to form a surface modulating assembly that regulates surface receptor mobility and cell growth. To test this hypothesis, we examined three agents known to affect cell growth [colchicine, concanavalin A (Con A), and the src gene product of Rous sarcoma virus] for their effects on chick embryo fibroblasts. Individual cells from serum-starved normal fibroblast populations became committed to enter S phase at various times over a 12 hr period after exposure to serum. Colchicine and other microtubule-disrupting agents blocked entry into S phase at a point close to the commitment point for each cell. The lectin Con A also blocked entry into the S phase when present in doses sufficient to modulate surface receptor mobility. In contrast, succinyl-Con A, which does not induce surface modulation, had no effect. Both Con A and colchicine blocked the appearance of cytoplasmic factors capable of stimulating DNA replication in a cell-free system. To study endogenous effects on the surface modulating assembly, we infected fibroblasts with a Rous sarcoma virus (tsNY68) having a temperature-sensitive mutation in the transforming (src) gene. We have previously shown that microtubular and microfilamentous structures of the surface modulating assembly are direct or indirect targets of the src gene product with consequent reduction in the capacity of Con A to induce surface modulation. TsNY68-infected fibroblasts shifted to the non-permissive temperature acquired normal microtubular morphology more rapidly (2 hr) than cells grown at the permissive temperature in the presence of protein synthesis inhibitors (7.5 hr). This suggests that the src gene product acts directly on the surface modulating assembly rather than via the nucleus or at the level of protein synthesis. Furthermore, "transformation" of the surface modulating assembly was partly blocked by treatment of the infected cells with Con A but not succinyl-Con A. Both Con A and colchicine inhibited entry into the S phase following a shift from nonpermissive to permissive growth conditions. All of these observations are in accord with the hypothesis that the surface modulating assembly acts as a signal regulator in growth control.

Topics: Animals; Avian Sarcoma Viruses; Cell Division; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Colchicine; Concanavalin A; DNA Replication; Fibroblasts; Time Factors

Topics: Agglutination; Animals; Bucladesine; Cell Line; Cell Transformation, Neoplastic; Cell Wall; Cells, Cultured; Clone Cells; Concanavalin A; Mice; Microscopy, Electron, Scanning; Theophylline

Using a series of cold-sensitive variants of chemically transformed BHK-21 cells, revertants to the normal phenotype derived from a dimethyl-nitrosamine transformed clone of BHK-21 as well as revertants to the normal phenotype derived from polyoma transformed BHK-21 cells we have demonstrated that the surface phenotype described by enhanced agglutinability with Con A and WGA can be dissociated from the transformed phenotype described by anchorage independence (growth in semisolid medium). Specifically we have demonstrated that the surface characteristic of enhanced agglutinability may be found in a variety of cell lines which fail to display to grow in agar. Our work clearly shows that the two phenotypes described are not concomitantly controlled and tends to suggest that the phenotype of enhanced lectin agglutinability may be dissociated from the transformed phenotype.

Topics: Agar; Agglutination; Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Clone Cells; Cold Temperature; Concanavalin A; Cricetinae; Lectins; Nitrosamines; Phenotype; Plant Lectins; Polyomavirus; Triticum

Topics: Adenosine; Adenosine Monophosphate; Bucladesine; Butyrates; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cyclic GMP; Receptors, Concanavalin A; Receptors, Drug; Theophylline; Trypsin

Topics: Agglutination; Animals; Avian Leukosis Virus; Bone Marrow; Cell Transformation, Neoplastic; Cell Transformation, Viral; Chick Embryo; Concanavalin A; Culture Techniques; Deoxyglucose; Fibroblasts; Hematopoietic Stem Cells; Lectins; Neoplasm Proteins; Peptide Hydrolases

AD6, a mutant derived from 3T3 Balb/c cells, is characterized by low adhesion to substratum, round shape, increase in surface microvilli, increase in agglutinability by concanavalin A, and loss of directional motility. These properties are often observed in transformed cells. However, the mutant has normal growth properties and anchorage-dependence of growth, and it does not form tumors. In AD6, the biosynthesis of complex carbohydrates and glycoproteins is impaired because of a block in the acetylation of GlcN-6-P. This defect is responsible for all the surface alterations because feeding of GlcNAc to AD6 cells corrects the defects in the synthesis of complex carbohydrates and the exposure of glycoproteins at the outer surface of the plasma membrane. Parallel to this biochemical reversion, there is full restoration of the altered biological properties. In contrast, GlcNAc has no effect on the morphologic features of two lines of transformed cells. Our results suggest that the carbohydrate portion of cell surface proteins has an important role in adhesion and related aspects of cell behavior. The fact that a defined alteration of the cell surface induces many properties often encountered in transformed cells, without affecting control of cell division, strongly suggests that these alterations in properties are not sufficient to account for the loss of growth regulation.

Topics: Acetylglucosamine; Cell Adhesion; Cell Aggregation; Cell Division; Cell Line; Cell Movement; Cell Transformation, Neoplastic; Concanavalin A; Fibroblasts; Genotype; Glucosamine; Glycoproteins; Membrane Proteins; Microscopy, Electron, Scanning; Mutation; Phenotype

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

The effects of Moloney Sarcoma Virus (MSV) induced tumor growth dynamics on the blastogenic responsiveness of lymphocytes from BALB/c mice were investigated. Lymphocytes from spleen, thymus and lymph node pools were tested for responsiveness to phytohemagglutinin (PHA) and concanavalin A (Con A). Results showed a significant decrease in PHA-induced blastogenesis of all lymphocytes tested at the time of maximal tumor volume, with a return to normal responsiveness as the tumor regressed. In contrast, a differential dose dependent Con A response occurred in spleen and thymus lymphocytes. A decreased 3H-thymidine uptake occurred at optimal Con A dose, correlating with the PHA decrease. However, at a lower Con A dose an increased response was observed, beginning shortly before the PHA depression and continuing until regression of tumor began. This phenomena was not observed in lymph node lymphocytes. Based upon these observations, we suggest that the cell or cells responsible for the transient suppression of PHA responsiveness may be Con A responsive T lymphocytes.

Topics: Animals; Cell Transformation, Neoplastic; Concanavalin A; Dose-Response Relationship, Immunologic; Leukemia, Experimental; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Moloney murine leukemia virus; Spleen; T-Lymphocytes

Topics: Agglutination; Calcimycin; Calcium; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fibroblasts; Metals

Topics: Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fatty Acids; Membrane Lipids; Mice; Phenotype; Temperature

Whereas Concanavalin A (Con A) and Wheat Germ Agglutinin (WGA) detect differences in the agglutinability of transformed, established and secondary cultures, Phytohemagglutinin (PHA) detects differences between cultured adult and fetal human fibroblasts. Adult cells agglutinate with PHA to the same extent as transformed cells, whereas fetal cells show significant agglutination only after trypsinization. Differences in cell size, growth rate, surface architecture or binding of fluorescent PHA could not be demonstrated between adult and fetal cells. Although the basis for this apparent difference in agglutinability remains unknown, it is the first demonstration that fetal cells (even after prolonged in vitro culture) retain at least some surface properties not shared by adult or transformed cells.

Topics: Adult; Agglutination; Animals; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fetus; Fibroblasts; Humans; Lectins; Mice; Plant Lectins; Rats; Triticum

Functional markers and growth behavior of abnormal hepatocytes at several stages of liver carcinogenesis were studied. Early lesions, i.e., hyperplastic foci and areas, did not accumulate iron in siderotic livers, had persistent glycogen stores, were not more agglutinable by concanavalin A, and were associated with alpha-fetoprotein secretion, but were not independent secretors of high amounts. The cells in the early lesions had an increased mitotic index, but cells from livers with early lesions did not have an increased survival in cell culture or the ability to grow in soft agar. The more developed lesions, hyperplastic nodules, also did not store iron, had persistent glycogen, did not display increased concanavalin A agglutinability, and were not independent secretors of high levels of alpha-fetoprotein. Similarly, nodule cells were proliferative but did not display an increase in survival in cell culture. In addition, both iso- and autotransplantation of nodules into mammary fat pads resulted in persistence but not growth of nodule cells. On the other hand, hepatocellular carcinomas regularly grew upon transplantation. Thus, early lesions and hyperplastic nodules were proliferative lesions did not possess autonomous growth capability comparable to that of hepatocellular carcinomas.

Topics: 2-Acetylaminofluorene; alpha-Fetoproteins; Animals; Cell Aggregation; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Hyperplasia; Iron; Liver Glycogen; Liver Neoplasms; Neoplasms, Experimental; Precancerous Conditions; Rats

The surface morphology of attached and suspended normal and transformed fibroblasts has been studied with the scanning electron microscope. Normal murine fibroblasts (3T3) grow in vitro with widely extended leading lamellae. During most parts of the cell cycle the surfaces of these cells are practically free of microvilli. When the cells round up for mitosis, their cell surfaces become adorned with many microvilli. In contrast, simian virus 40-transformed fibroblasts (SV3T3) grow more compact, and their cell surfaces remain smooth throughout the life cycle. When confluent 3T3 and SV3T3 cells are suspended with ethylenediaminetetraacetic acid (EDTA) for agglutination assays, similar differences in surface morphology are found: 3T3 cells always bear many microvilli, whereas most SV3T3 cells are essentially free of microvilli. The addition of concanavalin A (Con A) does not influence the surface morphology of the suspended cells. The morphological differences described here may be important for the agglutination process of the normal and transformed 3T3 cells, because they affect the real cell surface area and thus the density of Con A-binding sites.

Topics: Agglutination Tests; Cell Aggregation; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fibroblasts; Microscopy, Electron, Scanning; Simian virus 40; Time Factors

Topics: Animals; Base Sequence; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; DNA, Neoplasm; DNA, Viral; Mice; Neoplasm Transplantation; Polyomavirus

The extent of binding of 125I-Con A to the surface of SV40-transformed human fibroblasts and the degree of agglutination of the cells by the native lectin have been measured. In addition, trypsinized and succinylated Con A have been used to study the effects of the lectin upon certain cell growth parameters. Trypsinized Con A was found to alter the growth rate, the saturation density and the contact inhibition of the transformed cells, an effect not neutralized by alph-methyl-D-mannoside.

Topics: Agglutination; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fibroblasts; Humans; Mannosides; Simian virus 40

The fluorescence recovery kinetics of succinyl-fluorescein Concanavalin A (S-F-ConA) in glycerol-physiological saline solutions of high viscosity and when bound to the surface of mouse fibroblasts were measured following brief photobleaching using a laser excited fluorescence microscope. In the high viscosity solutions, the recovery kinetics, interpreted on the basis of a simple diffusion model, yielded a diffusion coefficient in close agreement with the values predicted by the Stokes-Einstein equation. Recovery kinetics for S-F-ConA bound to the surface of mouse 3T3 and SV3T3 cells cultured in vitro yielded diffusion coefficients in the range of 5-10-10(-11) cm2/s, values considerably lower than those reported previously for membrane proteins. These measurements indicated that a considerable fraction of the S-F-ConA molecules bound to the cell surface are immobilized. These results are discussed in relation to current concepts of lateral motion of protein components within natural membranes.

Topics: Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Glycerol; Kinetics; L Cells; Lasers; Mathematics; Protein Binding; Proteins; Simian virus 40; Sodium Chloride; Spectrometry, Fluorescence

Studies on amino acid and hexose transport were performed on human WI-38 cells and WI-38 SV40-transformed cells (VA13A and VA13-2RA). Depending upon cell line or conditions, either no difference or a relative decrease in initial uptake by transformed cells was found. Under similar growth conditions, transformed hamster cells (PyBHK-21/C13) had increased uptake, compared with the normal hamster cells (BHK-21/C13). The normal and transformed human cells were also similar in sialic acid content and agglutinated when treated with concanavalin A.

Topics: Agglutination Tests; Aminoisobutyric Acids; Animals; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Deoxy Sugars; Deoxyglucose; Humans; Sialic Acids; Simian virus 40

Topics: Blood; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; Fibrinolysin; Plasminogen; Simian virus 40

We demonstrated that the productive infection of three different mammalian cell lines with two separate leukemia viruses is sufficient to induce a change in surface architecture that may be detected as enhanced agglutinability with two different plant lectins. Subsequent transformation of one of these cell lines with a chemical carcinogen did not further modify the agglutinability of the cell lines. Using a polyoma virus-transformed derivative of one of the parental lines, we have demonstrated that the LETS protein (whose absence from the surface membrane has been considered a marker of the transformed phenotype) may be present in cells displaying the capacity to plate in soft agar.

Topics: Agar; Agglutination; Animals; Carcinogens; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Glycoproteins; Iodine Radioisotopes; Lactoperoxidase; Lectins; Leukemia Virus, Murine; Molecular Weight; Neoplasm Proteins; Nitroquinolines; Rodentia; Surface Properties

Seven lines derived from primary African green monkey kidney cells, which had survived lytic infection by wild-type simian virus 40 (SV40) or temperature-sensitive mutants belonging to the A and B complementation groups, were established. These cultures synthesize SV40 tumor (T) antigen constitutively and have been passaged more than 60 times in vitro. The cells released small amounts of virus even at high passage levels but eventually became negative for the spontaneous release of virus. Virus rescued from such "nonproducer" cells by the transfection technique exhibited the growth properties of the original inoculum virus. Four of the cell lines were tested for the presence of altered growth patterns commonly associated with SV40-induced transformation. Although each of the cell lines was greater than 99% positive for T antigen, none of the cultures could be distinguished from primary or stable lines of normal simian cells on the basis of morphology, saturation density in high or low serum concentrations, colony formation on plastic or in soft agar, hexose transport, or concanavalin A agglutinability. However, the cells could be distinguished from the parental green monkey kidney cells by a prolonged life span, the presence of T antigen, a resistance to the replication of superinfecting SV40 virus or SV40 viral DNA, and, with three of the four lines, an ability to complement the growth of human adenovirus type 7. These properties were expressed independent of the temperature of incubation. These results indicate that the presence of an immunologically reactive SV40 T antigen is not sufficient to ensure induction of phenotypic transformation and suggest that a specific interaction between viral and cellular genes and/or gene products may be a necessary requirement.

Topics: Adenoviridae; Agglutination; Animals; Antigens, Viral; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Deoxyglucose; Haplorhini; Mutation; Simian virus 40; Temperature; Virus Replication

Cell membranes from mouse L-cells (L-B82), rat hepatoma (HTC-H1), and three clones of their somatic cell hybrids (07, V4a, and V5) showing different degrees of density-dependent inhibition of growth were analyzed by polyacrylamide gel electrophoresis. The membrane polypeptides of the hybrid clones were all similar and all showed higher proportions of polypeptides with molecular weights of 56,000 and 45,000 than their parents of their normal counterparts. The major glycoprotein form cell hybrids appeared to be identical with that of rat liver or rat hepatoma cells and different from that of L-cells. One hybrid showed density-dependent inhibition growth; the other two, like both parents, did not. All produced tumors in nude mice, although tumor production by the hybrids was delayed. A large external protein (M.W. 240,000) iodinated by lactoperoxidase-catalyzed reaction was virtually missing in the parents but was present at high levels in all their hybrid clones. Thus, there was a lack of correlation between the presence of this protein, growth control in vitro, and tumorigenicity. Furthermore, no correlation was seen between agglutination of these cells by concanavalin A and tumorigenicity. The factors controlling these membrane properties thus are independent of density-dependent inhibition of growth and of those controlling the expression of cancer.

Topics: Agglutination; Animals; Carcinoma, Hepatocellular; Cell Fusion; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Glycoproteins; Hybrid Cells; L Cells; Lactoperoxidase; Liver Neoplasms; Mice; Molecular Weight; Neoplasms, Experimental; Peptides; Rats

The hypothesis that surface modulating assemblies containing microfilaments and microtubules and altered after cellular transformation was tested on cells infected with temperature-sensitive mutants of avian sarcoma virus. Untransformed cells (mouse 3T3 and chick fibroblasts), cells transformed by simian virus 40 (SV 3T3), and chick fibroblasts infected with Schmidt-Ruppin strain of Rous sarcoma virus (SR-RSV-A-infected cells) were first compared for differences in microfilament and microtubule patterns after treatment with fluorescein-labeled antibodies to actin and tubulin. Transformed cells showed disappearance of ordered stress microfilaments and thickened or diffuse alterations of microtubular arrays. At restrictive temperatures (41 degrees), chick fibroblasts infected with a temperature-sensitive mutant (ts 68) of Rous sarcoma virus showed normal patterns of stress fialments and radial microtubular arrays originating in 1 or 2 centrioles. At permissive temperatures (37 degrees), these patterns were disordered and resembled those of SR-RSV-A-infected cells. After a shift from 41 degrees to 37 degrees, the changes in microtubules were observed in the majority of cells within 1 hr. These changes were reversible and did not result from the inability of tubulin to polymerize. In ts 68-infected cells at permissive temperatures, concanavalin A induced much less surface modulation (inhibition of receptor mobility) than at restrictive temperatures. These results suggest that cellular transformation alters both the structure and function of surface modulating assemblies and prompt the hypothesis that products of viral transforming genes may affect these assemblies with a consequent loss of growth control.

Topics: Actins; Avian Sarcoma Viruses; Cell Transformation, Neoplastic; Concanavalin A; Microtubules; Mutation; Surface Properties; Temperature; Tubulin

Topics: Animals; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; Centrifugation; Concanavalin A; Culture Media; Dextrans; Lectins; Liver Neoplasms; Lymphocyte Activation; Neoplasm Transplantation; Rats; Time Factors

Topics: Age Factors; Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cytotoxicity Tests, Immunologic; Histocompatibility Antigens; Immunity, Cellular; Male; Mice; Simian virus 40; T-Lymphocytes; Teratoma; Testis

Polyoma-transformed (Py-transformed) BHK-21 cells are agglutinated more readily by Concanavalin A (Con A) after they are infected with influenza virus; this agglutination was only slightly affected by fixation with glutaraldehyde or colchicine treatment. It is suggest that the mechanism is not dependent on membrane fluidity. The same amounts of 125i-labelled Con A were bound after infection with influenza virus and after glutaraldehyde fixation with or without colchicine treatment: it seems therefore that new receptors for Con A are not expressed between 90 and 600 minutes after these cells are infected with influenza virus.

Topics: Agglutination; Binding Sites, Antibody; Cell Line; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Erythrocytes; Glutaral; Hemadsorption; Humans; Iodine Radioisotopes; Orthomyxoviridae; Polyomavirus

Cell strains isolated from an established line of SV40-transformed mouse 3T3 cells (SV3T3) showed large variations in the various parameters of transformation, viz. saturation density, serum requirement, contact inhibition of movement and of growth and Concanavalin-A-mediated agglutinability. These cell strains were studied for changes in elution profiles of fucose-labelled surface glycoproteins, using actively growing, untransformed 3T3 cells as controls. A cell strain established from a tumour arising after injection of wild-type SV3T3 cells and SV3T3 cells grown in vivo in diffusion chambers, was similarly studied. Changes in surface glycoproteins were observed only in the tumour-derived cell strain. The results suggest that alterations in surface glycoproteins are associated with the tumorigenic potential of the cells rather than with the transformed phenotype per se.

Topics: Agglutination; Animals; Cell Aggregation; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; DNA; Glycoproteins; Mice; Simian virus 40

Normal and premalignant mouse mammary epithelial cells can be prepared in high yields by collagenase dissociation of minced glands followed by a brief, differential centrifugation to remove contaminating fibroblasts and fat cells. The major difficulties in preparing pure cultures in quantity are 1) incomplete dissociation of gland material, and 2) cell death during enzymatic digestion. These problems are eliminated by careful selection of collagenases for dissociation. Normal and premalignant mammary epithelial cells are morphologically indistinguishable from malignant mouse mammary epithelial cells in primary monolayer cultures. In addition, the growth rates and saturation densities achieved by normal mammary epithelial cells are indistinguishable from those of malignant mammary epithelial cells in primary culture. In both cases, a monolayer of cells is preserved with no evidence of focal overgrowth. Malignant adenocarcinoma mammary cells can however be distinguished from normal mammary epithelial cells by virtue of differences in their surface interactions with concanavalin A. A hemadsorption assay using Con-A-coated erythrocytes was the most sensitive indicator for these differences. In hemadsorption assays malignant mammary epithelial cells were half-maximally reactive with 2.5 mug/ml concanavalin A, while normal cells were completely unreactive even at concanavalin A concentrations five-times higher. Premalignant mammary epithelial cells were as reactive as malignant mammary epithelial cells in the hemadsorption assays. Hemadsorption of malignant cells was observed in primary and secondary cultures of epithelium as well as in cell lines. Malignant cells forming mammary adenocarcinomas were as highly reactive as malignant cells forming scirrhous carcinomas. Malignant cells not releasing mammary tumor virus (MuMTV) were as reactive as cells releasing that virus. Adsorption of concanavalin-A-coated erythrocytes to normal mammary epithelial cells could be induced by brief treatment of cell monolayers with hyaluronidase. Exposure of active sites was not affected with either trypsin or collagenase. Our results show that while the growth of malignant cells does not serve to distinguish them from normal cells in monolayer culture, surface changes do exist which can be identified by differences in concanavalin A reactivity. Since the earliest transformants identifiable in vivo (premalignant) have undergone conversion of the surface marker, concanavalin-A-medi

Topics: Animals; Autoradiography; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Epithelial Cells; Epithelium; Female; Hemadsorption; Hyaluronoglucosaminidase; Mammary Neoplasms, Experimental; Mice; Mice, Inbred Strains; Microbial Collagenase; Trypsin

Concanavalin A (Con A) concentration dependence of agglutinability of a mutant of Rous sarcoma virus which is temperature-sensitive for transformation was examined. Con A agglutinability of the virion was quantitatively expressed by measuring radioactivities of 60 to 70 S RNA extracted from Con A-agglutinated material. The mutant grown at a permissive temperature (35 degrees C) agglutinated at a significantly lower concentration of Con A, compared with that at a non-permissive temperature (40 degrees C).

Topics: Agglutination; Avian Sarcoma Viruses; Cell Transformation, Neoplastic; Concanavalin A; Humans; Infant, Newborn; Mutation; RNA, Viral; Temperature

The interactions between concanavalin A and chick embryo fibroblasts, normal and infected with Rous sarcoma virus (RSV-BH) or its thermosensitive mutant RSV-BH-Ta, have been studied. Normal chick embryo cells and RSV-BH transformed cells showed at 4 and 25 degrees C a similar number of concanavalin A receptors per cell. Analysis of the binding data by the Scatchard relation showed that apparent changes in binding as a function of temperature are due to the thermodynamic properties of the process and not to endocytosis. The lectin receptors on the cell surface of normal and RSV-BH infected cells showed homogeneity in their binding properties. Chick cells infected with RSV-BH-Ta showed a lectin binding behavior that was dependent on the temperature at which the cells were grown. At the permissive temperature for transformation (37 degrees C), the binding process was similar to that observed for normal and RSV-BH infected cells. At the nonpermissive temperature (41 degrees C), the cells showed at least two sets of concanavalin A receptors. The new set of receptors on the cell surface had a lower lectin affinity than those observed in the same cells at 37 degrees C. Chick cells infected with RSV-BH showed an enhanced agglutinability by concanavalin A, as compared with normal cells. Cells infected with RSV-BH-Ta showed a reversal of the correlation between increased concanavalin A agglutinability and the transformed state. At the permissive temperature for transformation, the cells were not agglutinable, whereas at the nonpermissive temperature they presented agglutinability indexes as high as those observed with RSV-BH infected cells. This enhanced agglutinability observed with cells maintained at the nonpermissive temperature for transformation may be related to the new set of low affinity receptors present at 41 degrees C.

Topics: Agglutination Tests; Animals; Avian Sarcoma Viruses; Binding Sites; Binding, Competitive; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Fibroblasts; Kinetics; Methylmannosides; Mutation; Temperature; Thermodynamics

A variant of polyoma BHK cells is described which is capable of growing in a glutamine-free medium. These glutamine-independent variant cells (GIV) have a size and morphologic appearance intermediate between the nontransformed (BHK) and polyoma transformed (Py6) cell line. In contrast to Py6, the GIV cells grow more slowly, produce less lactic acid and do not grow in suspension. However, their adherence to substrate and release of galactosyltransferase into the medium is comparable to that seen with the parent PyBHK. Most striking is the finding that GIV cells are more readily agglutinated by Concanavalin A and when inoculated into hamsters are much more tumorigenic than the parent Py6 cells.

Topics: Agglutination; Animals; Cell Adhesion; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Galactosyltransferases; Genetic Variation; Glutamine; Lactates; Neoplasms, Experimental; Polyomavirus

Scanning electron microscope studies were carried out on Syrian hamster embryo cells transformed in vitro by X-irradiation (300 rads) (X-ray transformed) and on normal nonirradiated and irradiated nontransformed controls. Transformed cells appeared in scanning electron microscopy as pleomorphic, thick cells piling up over each other and exhibiting extensive surface features consisting of microvilli, blebs, and ruffles. These surface structures were seen on single as well as on densely cultured transformed cells during both interphase and mitosis. The complex surface was observed shortly after transformation (on cells of a 20-day-old clone) and seems a permanent feature of the X-ray-transformed cells (present after 8 years in culture). All controls appeared by scanning electron microscopy as regular, flat, and smooth cells which grew in high-density cultures to seemingly contact-inhibited monolayers. During mitosis the normal cells (control, nontransformed) displayed surface excrescences similar to those of the transformed cells making the mitotic normal cells indistinguishable from transformed cells. The complex surface features in the normal cells were temporary and reversed back to characteristic smoothness upon reentrance into interphase.

Topics: Agglutination; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Clone Cells; Concanavalin A; Contact Inhibition; Lectins; Microscopy, Electron, Scanning; Microscopy, Phase-Contrast; Mitosis; Radiation Effects; X-Rays

Topics: Agglutination; Animals; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Erythrocytes; Lectins; Leukemia, Experimental; Male; Mice; Mice, Inbred Strains; Neuraminidase; Osmotic Fragility; Polylysine; Rauscher Virus

Agglutinability by concanavalin A, distribution of surface-bound concanavalin A, and maximal cell density in monolayer culture were examined under similar conditions in parallel cultures of ten established cell lines. The degree of agglutinability of the cell lines did not correlate with the presence or absence of patching of concanavalin A bound to the cell surface, as determined with a hemocyanin marker. Agglutinability was also not always correlated with the loss of post-confluence inhibition of cell division. Two clones of mouse 3T3 fibroblasts that maintained post-confluence inhibition of cell division and low agglutinability differed substantially with respect to the surface distribution of concanavalin A. Patching of concanavalin A binding sites is neither necessary nor sufficient to explain differences in agglutinability between cell lines.

Topics: Agglutination; Binding Sites; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition

Topics: Agglutination; Bronchial Neoplasms; Cell Transformation, Neoplastic; Cells, Cultured; Chromosomes; Concanavalin A; Fibroblasts; Humans; Male; Middle Aged; Mitosis

Topics: Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Herpesviridae; Methylmannosides; Trypsin

Topics: Agglutination; Binding Sites; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Genetic Variation; Kinetics; Temperature; Trypsin

Topics: Acetylglucosamine; Agglutination; Cell Transformation, Neoplastic; Concanavalin A; Dinitrofluorobenzene; Fixatives; Granulocytes; Humans; Leukocytes; Methylglucosides; Nitrobenzenes; Receptors, Concanavalin A; Receptors, Drug; Serum Albumin, Bovine

Clones of four Gross virus-induced murine lymphoblast lines, established in culture from C3H mice, were selected for detailed study of the relationships among in vitro growth parameters, oncogenicity, and agglutination with concanavalin A. The four clones were intially divided into two groups on the basis of their in vitro growth properties. Two strains, N-811 and H-111, had low saturation densities, low cloning efficiencies, and slower doubling times; the other two strains, L-274 and L-258, had higher saturation densities, higher cloning efficiencies, and faster doubling times. The ability of the strains to produce tumors in mice correlated with their in vitro growth properties: L-274 and L-258, with their high saturation densities and high cloning efficiencies, were more tumorigenic in mice than were N-811 and H-111 cells with their lower saturation densities and lower cloning efficiencies. All strains were agglutinable with concanavalin A; however, the agglutination response did not correlate with saturation density or oncongenicity.

Topics: Agglutination; AKR murine leukemia virus; Animals; Cell Division; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Leukemia, Experimental; Mice; Mice, Inbred C3H; Neoplasm Transplantation; Transplantation, Isogeneic

Transmission and scanning electron microscopy were used to study possible structural correlates in the process of agglutination of several types of normal and transformed cells by Concanavalin A. In parallel studies we found that post-confluence inhibition of cell division and agglutiniability of cells by Concanavalin A were not correlated with patching of surface bound lectin molecules as determined with a hemocyanin marker. Transformed cells growing in monolayer cultures were found to have many more microvilli than the corresponding normal cells. However, when cells were brought into suspension with EDTA, all cells developed numerous microvilli and we were not able to distinguish between agglutinable and nonagglutinable cells on the basis of morphological appearance. Cells agglutinated by Concanavalin A had numerous interdigitated microvilli at points of cell-cell contact. The appearance of spontaneously agglutinated cells and lectin agglutinated cells was very similar with respect to the involvement of microvilli in cell-cell attachments, and labeling studies with hemocyanin indicated that Concanavalin A bound to microvilli is rapidly cleared from these surface specializations in a manner analogous to that observed with patching of surface bound lectin. Several lines of SV-40 transformed fibroblasts were shown to be considerably more spontaneously agglutinable than untransformed cells. These results indicate that Concanavalin A may amplify an intrinsic membrane property common to many transformed cells that is expressed as an increase in the rate of adhesion of suspended cells. It is proposed that the membrane change detected by the agglutination reaction may also be involved in the loss of post-confluence inhibition of cell division and growth of transformed cells in semisolid media, due to a surface interaction that allows transformed cells to use each other as growth substrata.

Topics: Agglutination; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; Hemocyanins; Microscopy, Electron; Microscopy, Electron, Scanning; Protein Binding

The infection of isolated B and T cells by a murine leukemia virus (Friend) MuLV-F) was studied both in vitro and in vivo with an implanted diffusion chamber system. Lymphocytes were obtained from pools of normal spleen cells by filtration of the cell suspension through a nylon-wool column. The purity for both Ig positive and theta-positive cells varied between 85% and 90% in the B-cell and T-cell fractions; both lymphocyte fractions responded very well to stimulation with their respective specific polyclonal mitogens, bacterial lipopolysaccharide (LPS) and concanavalin A (Con A). Lymphocytes were infected by incubating pelleted cells in 2-6 x 10(4) FFU MuLV for 1 h at 4 degrees C and were then cultured for 5-10 days. Cells releasing infectious MuLV were enumerated as infectious centers (IC). IC were really detectable in the cultures of infected B-cells but none were found in the T-cell cultures. Addition of LPS to the culture medium increased the number of IC in B-cell fractions up to 1,000-fold. Furthermore, in T-cell cultures with LPS, IC also appeared in number which approximately correlated with the contaminating Ig+ cells of the T-cell fraction. In contrast, Con A had no consistent effect on the infection of either B or T cells. In the absence of MuLV-F, mitogenic stimulation alone did not elicit any endogenous IC. In subsequent experiments, purified lymphocytes were infected in diffusion chambers in vivo. The number of IC in infected B cells increased 1,000-fold as compared to infection in tissue culture. The peak of infection at 10 days was followed by a slight decline. Infected cells were also found in diffusion chambers containing T-cell fractions; these IC had very similar kinetics to those in B-cell-containing chambers, but their number was 10 times lower, suggesting that the infected cells were B cells, which comprised about 10% of the T-cell fraction. The virus-related antigens were detectable by immunofluorescence on the membrane of cells recovered from B-cell-bearing chambers but not on cells from T-cell-bearing chambers.

Topics: Animals; Antigens, Viral; B-Lymphocytes; Bone Marrow; Bone Marrow Cells; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Female; Friend murine leukemia virus; Leukemia, Experimental; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Mitogens; Polysaccharides, Bacterial; Spleen; T-Lymphocytes; Virus Replication

The sorting-out behavior (self-segregation of two cell types from mixtures of the two) of five different established cell lines was studied. Eight of the ten possible binary combinations of these lines, cultured as cellular aggregates, were examined. Mouse BALB/c 3T3 cells sorted out internally to the corresponding malignant SV40 virus-transformed 3T3 cells. The transformed 3T3 line (SVT-2) did not sort out from a revertant line selected from SVT-2 cells by resistance to concanavalin A (con A). The revertant cells sorted out externally to the parent BALB/c 3T3 cells, although segregation was generally incomplete. BALB/c 3T3 cells did not sort out from another contact-inhibited line of 3T3 cells derived from Swiss albino mice (Swiss 3T3). Both BALB/c 3T3 and Swiss 3T3 cells sorted out from cells of the contact-inhibited hamster line, NIL B. Instead of a two-layered sphere, however, a three-layered structure was observed with most of the NIL B cells external to the 3T3 cells, and a few NIL B cells comprising the center of the sphere. On the other hand, NIL B cells did not consistently sort out from either the SVT-2 or con A cells. In general, sorting out between pairs of these five lines are slower and less complete than is generally observed between the more extensively studied chick embryonic tissue cells, suggesting that the cultured cells may be more closely related in their adhesive properties. The internal segregation of BALB/c 3T3 cells relative to SVT-2 cells is consistent with the hypothesis that transformed cells are less adhesive than their nontransformed counterparts.

Topics: Cell Adhesion; Cell Aggregation; Cell Line; Cell Separation; Cell Survival; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition

Topics: 3,3'-Diaminobenzidine; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Ferritins; Fibroblasts; Hemocyanins; Histocytochemistry; Mice; Mice, Inbred BALB C; Peroxidases; Simian virus 40; Staining and Labeling

Human lymphoid cells isolated from the peripheral blood of adults, from cord blood, and from fetal liver, spleen, bone marrow, and thymus were cultivated with or without a cell-free preparation of Epstein-Barr virus (EBV) with demonstrated transforming activity. The cultures were examined for the EBV-associated nuclear antigen (EBNA) and for transfromation into permanent lymphoblastoid cell lines (LCL). EBNA, seen only in cultures that had received exogenous EBV, was detected between days 1 and 6 after addition of EBV, most frequently on day 3. EBNA-positive cells had a lymphoblastoid appearance. Transformation into established LCL became apparent between days 12 and 19. The addition of pokeweed mitogen to cultures containing EBV enhanced the development of EBNA, whereas phytohemagglutinin or concanavalin A had no such effect. Neither EBNA nor transfomration was observed in lymphoid cells from fetal thymus. In fetal spleen, bone marrow, and liver cells, EBV regularly induced EBNA and LCL transformation.

Topics: Antigens, Viral; Blood; Bone Marrow; Cell Line; Cell Nucleus; Cell Transformation, Neoplastic; Concanavalin A; Herpesvirus 4, Human; Humans; Lectins; Liver; Lymphocytes; Spleen; Thymus Gland; Time Factors; Umbilical Cord; Virus Replication

Topics: Agglutination; Animals; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Fibroblasts; Mice; Polyomavirus; Surface Properties

Hamsters vaccinated with adenovirus-transformed cells, modified by acetoacetylation or concanavalin A treatment, or with small numbers of living cells were partly or completely protected against challenge with 3 times 10-6 living cells. Treatment of vaccine cells with iodoacetate, Mitomycin C, neuraminidase plus Mitomycin C did not produce efficient vaccines. Herpes simplex virus-transformed cells treated by any of these procedures did not prevent, and frequently even enhanced, the growth of the homologous living cells; enhancement was often greater in female than in male hamsters. Protective and enhancing vaccines did not induce a different level of cell-mediated immunity, as detected by lymphocytotoxicity tests, which were positive for both homologous transformed cells and nontransformed hamster cells. In contrast, specific complement-dependent cytotoxic antibodies active only on adenovirus-transformed cells were induced by the protective acetoacetylated vaccine prepared from adenovirus-transformed cells; these antibodies were not present after nonprotective vaccinations. The appearance of herpes simplex virus tumors was delayed by treatment with the immunostimulant, Levamisole, or by preimmunization with Newcastle disease virus grown in SV40-transformed cells, but not by Newcastle disease virus grown in herpes simplex virus-transformed cells. Thus, only nonspecific treatments were able to impede herpes simplex virus tumor growth, while protection against adenovirus tumor was accompanied by specific cytotoxic antibodies.

Topics: Acetylation; Adenoviridae; Animals; Antibodies, Viral; Antibody Formation; Antigen-Antibody Complex; Antigens, Neoplasm; Cell Transformation, Neoplastic; Complement System Proteins; Concanavalin A; Cricetinae; Cytotoxicity Tests, Immunologic; Female; Immunity, Cellular; Iodoacetates; Levamisole; Lymphocytes; Male; Mitomycins; Neoplasms, Experimental; Neuraminidase; Newcastle disease virus; Simplexvirus; Time Factors; Vaccination

A 35SO4-labeling/chromatography technique has been developed which facilitates quantitation of sulfated glycosaminoglycan (GAG) synthesis in mammalian cell cultures. The technique has been used to compare sulfated GAG biosynthesis, degradation, and turnover in three related cell lines with differing degrees of density-dependent inhibition of growth in vitro (Balb/c 3T3, SV3T3, and SV3T3 revertant cells). Viral transformation of Balb 3T3 cells is accompanied by a 2-5-fold decrease in cell associated sulfated GAG. SV3T3 revertant cells, which show partial reversion to low saturation density in vitro, show a 2.5-8-fold increase in cell-associated sulfated GAG compared to the parental SV3T3 cells from which they were selected. In addition, the distribution of 35SO4 and [3H]glucosamine among the different GAG species produced by SV3T3 revertant cells reverts so that it is similar to the distribution characteristic of untransformed 3T3 cells rather than SV3T3 cells. Mild trypsin treatment of 35SO4-labeled cells removed 68-84% of the cellular sulfated GAG, suggesting that at least this proportion of the total cellular sulfated GAG was located at the cell periphery. Removal of 35SO4-labeled cells from the Petri dish with a Ca2+ selective chelating agent revealed a fraction of the sulfated GAG that remained tightly bound to the Petri dish. A higher proportion of the total cell-associated sulfated GAG remained attached to the Petri dish in cultures of untransformed and revertant cells compared to that present in cultures of transformed cells. A role for sulfated GAG in density-dependent growth inhibition of fibroblast cultures is proposed and discussed in the light of the data obtained.

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Chelating Agents; Concanavalin A; Glucosamine; Glycosaminoglycans; Mice; Mice, Inbred BALB C; Simian virus 40; Trypsin

Balb/c 3T3, SV40-transformed 3T3 (SVT2), and Con A revertant variants of transformed cells leave a layer of glycoprotein on the culture substrate upon EGTA mediated removal of cells. The metabolic properties of this substrate-attached material (glycoprotein) have been examined. Pulse and cumulative radiolabeling experiments with glucosamine and leucine precursors established that this substrate-attached material accumulates on the substrate in growing cultures until cells have completely covered the substrate. The synthesis and/or deposition of the material diminished dramatically in cultures whose substrates had been completely covered with cells as observed microscopically, even though the contact-inhibited cell lines continued to make cell-associated and medium-secreted glycoproteins and transformed cells continued to divide and form multilayered cultures. Pulse-chase analysis using long periods of pulsing with radioactive leucine demonstrated that these glycoproteins are deposited directly on the substrate by cells and not subsequent to secretion into the medium. The substrate-attached material accumulated during long pulses was stably adherent to the substrate and displayed little appreciable turnover during 3 days of chasing of either sparse or dense cultures. Short-term pulse-chase analysis with leucine revealed two metabolically different pools of material-one which turns over very rapidly with a half-life of 2-3 hr (observed in both low-density and high-density cultures) and a second pool which is stably deposited on the substrate and whose proportion increased with the length of the radiolabeling period. No appreciable differences in the metabolic properties of substrate-attached material were observed in the three cell types studied during growth on a plastic substrate. These results are discussed with regard to the implicated roles of these glycoproteins in in mediating adhesion of normal and virus-transformed cells to the substrate.

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Glucosamine; Glycoproteins; Leucine; Mice; Mice, Inbred BALB C; Simian virus 40; Time Factors

A bromodeoxyuridine(BUdr) dependent cell line, called B4, Which requires BUdr not only for optimal growth but also for the maintenance of the non-contact inhibited state was described previously. We have now shown that contact inhibition in the B4 cells in the absence of BUdr is associated with a marked decrease in the percent of cells synthesizing DNA. The transition to the contact inhibited state in the absence of BUdr does not seem to be due to changes in cyclic AMP levels. It has also been shown that several but not all of the characteristics which distinguish transformed from untransformed cells also distinguish B4 cells grown in the presence of BUdr from B4 cells grown in the absence of BUdr. In addition to being contact inhibited, B4 cells grown in the absence of BUdr have a higher serum requirement, grow less well in soft agar, and are less agglutinable by wheat germ agglutinin than B4 cells grown in the presence of BUdr. Agglutinability by concanavalin A, however, is the same for B4 cells grown in the presence and absence of BUdr. Dependent cells maintained in the presence of BUdr do not form tumors and it is not yet clear how the transformed characteristics of the dependent cells are related to malignancy.

Topics: Agglutination Tests; Animals; Blood Proteins; Bromodeoxyuridine; Cattle; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition; Cricetinae; Culture Media; Cyclic AMP; DNA, Neoplasm; Female; Lectins; Melanoma; Mutation; Neoplasm Transplantation; Plant Lectins; Triticum

Freeze-fracture techniques were used to study the ultrastructural distribution of plasma membrane particles in cultures of normal Balb/c and Swiss 3T3 and simian virus 40- or murine sarcoma virus-transformed fibroblasts. No apparent differences were observed. Cultures fixed in situ show a seemingly random distribution of membrane particles both in normal or in transformed cells. Treatment of cell cultures in situ with concanavalin A does not result in an altered pattern of particle distribution. EDTA-induced suspension of normal or transformed cells does not result, per se, in modification of the type of membrane particle distribution seen in cells fixed in situ. However, upon further incubation, a proportion of normal or transformed cells in suspension show a varying degree of particle aggregation following a network pattern. Concanavalin A treatment of normal and transformed cells in suspension does not result in a specific change of the pattern of particle distribution. Because it has been established that treatment with concanavalin A of simian virus 40-transformed Balb/c 3T3 fibroblasts causes pronounced clustering of the concanavalin A receptors at the outer-surface, our results probably imply independence of membrane particles and concanavalin A receptors of these transformed cells.

Topics: Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Fibroblasts; Freeze Etching; Gammaretrovirus; Intercellular Junctions; Mice; Mice, Inbred BALB C; Simian virus 40

The synthesis of proteins with affinity for DNA has been studied in clones of a Syrian hamster cell line (NIL) and subclones of this line transformed by polyoma virus (NIL-Py) or hamster sarcoma virus (NIL-HSV). The results show that the synthesis of DNA-binding proteins in NIL and in its virus-transformed derivatives NIL-Py and NIL-HSV is very similar in exponentially growing cells, but in dense culture there is a very significant difference in the level of a protein (P8), which is much higher in the transformed lines than in untransformed NIL. The high levels of P8 in dense transformed cells have been observed in all the clones of transformed cells examined, indicating that this behavior of P8 is related to transformation and not simply due to a fortuitous clonal selection from the NIL. Experiments with synchronized cells indicate that the time of maximal P8 synthesis relative to cellular DNA synthesis in NIL-HSV precedes that observed in NIL cells. P8 has a molecular weight of 30,000 as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and is present in large amounts in the transformed cells in dense culture, where it makes up 0.5 to 1% of the total soluble protein.

Topics: Agglutination Tests; Animals; Cell Line; Cell Transformation, Neoplastic; Chromatography, Affinity; Clone Cells; Concanavalin A; Cricetinae; DNA; Electrophoresis, Polyacrylamide Gel; Oncogenic Viruses; Polyomavirus; Protein Binding; Proteins; Receptors, Drug; Sarcoma, Experimental; Thymidine; Time Factors

We have utilized dark field microscopy to observe the surface microstructure of living cultured cells. Using this method, we have found that dibutyryl cAMP treatment causes regression of the numerous, long cell surface microvilli present on L929 cells. Thirty minutes after removal of dibutyryl cAMP, microvilli reappear. An inhibitor of phosphodiesterase (methylisobutylxanthine) and a stimulator of adenylate cyclase (prostaglandin E1), both of which raise cAMP levels, cause regression of microvilli in 15 min. Untransformed 3T3 cells show very few microvilli when viewed still attached to their substratum or after removal with EDTA. Treatment of these cells with trypsin causes the formation of numerous microvilli on their surface. When clumps of cells agglutinated by concanavalin A are examined by thin section electron microscopy, the cells are seen to be held together by a "forest" of interdigitating microvilli and only rarely is there apposition of the areas of membrane between microvilli. At the same time the distribution of surface-bound concanavalin A was examined using immunofluorescent light microscopy, and concanavalin A was found to be uniformly distributed over the cell surface. We propose that agglutinability of mouse and rat fibroblasts is regulated through the modulation of cell surface microvilli by cAMP, and that transformed cells are highly agglutinable because their low cAMP levels result in the formation of numerous surface microvilli.

Topics: Agglutination; Agglutination Tests; Animals; Bucladesine; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cyclic AMP; Fibroblasts; Fluorescent Antibody Technique; Mice; Microscopy, Electron; Microscopy, Phase-Contrast; Prostaglandins E; Time Factors; Xanthines

A quantitative hemadsorption assay distinguishes the effects of membrane fixation on concanavalin A-mediated agglutinability of fixed cells with unfixed cells. We observed undiminished adherence of unfixed erythrocytes to glutaraldehyde-fixed normal and virus-transformed hamster fibroblasts coated with concanavalin A. Fixation of the erythrocytes abolished agglutination with fixed fibroblasts. The agglutinability of fixed cells is more likely related to increased cell rigidity than to decreased membrane fluidity.

Topics: Agglutination; Animals; Cell Adhesion; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Cytological Techniques; Embryo, Mammalian; Erythrocytes; Fibroblasts; Glutaral; Hemadsorption Inhibition Tests; Hemagglutination Tests; Polyomavirus

On the basis of the previous study, on the cell interaction between malignant tumor cells and other cells, especially with lymphocytes, the present study was carried out by investigating cell to cell interaction of human malignant tumor cells and human lymphoblastoid cells such as T-cell (MOLT-4 cell) and B-cell (Burkitt lymphoma cell). As a result it has been revealed that live lymphoblastoid cells were not adhered on the cell surface of the tumor cells, nor is it ingested by tumor cells, but in thepresence of HVJ (Sendai virus: 2,000 H.A. units) it adheres slightly on the cell surface of tumor cell but no cell fusion of tumor cells and lymphoblastoid cells is observable. On the other hand, the tumor cell as well as T-cell and B-cell all have receptors to concanavalin A (Con. A) on their cell surfaces, and they show a marked cell binding such as tumor cell and T-cell, tumor cell and B-cell, and there can be observed a marked phagocytosis of lymphoblastoid cells by tumor cells. Moreover, the tumor cells that have phagocytized lymphoblastoid cells undergo a marked cell destruction within 4 hours of cell-binding and phagocytosis, which is especially prominent in the case of phagocytosis of E.B cell by tumor cell.

Topics: Animals; Antigen-Antibody Reactions; B-Lymphocytes; Cell Line; Cell Transformation, Neoplastic; Chickens; Concanavalin A; Cricetinae; Cytotoxicity Tests, Immunologic; Erythrocytes; Female; Glutaral; Humans; Immune Adherence Reaction; Leukemia, Lymphoid; Lupus Erythematosus, Systemic; Lymphocytes; Neoplasms; Ovarian Neoplasms; Parainfluenza Virus 1, Human; Phagocytosis; Sheep; T-Lymphocytes; Teratoma

Topics: Acetylglucosamine; Agglutination; Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Dose-Response Relationship, Drug; Fibroblasts; Glycine max; Kinetics; Lectins; Methylmannosides; Neuraminidase; Plant Lectins; Simian virus 40; Temperature; Triticum; Trypsin

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Dose-Response Relationship, Drug; Fibroblasts; Kidney; Lectins; Polyomavirus; Thymidine; Tritium

The ability of cells to bind to nylon fibers coated with lectin molecules interspaced with varying numbers of albumin molecules has been analyzed. The cells used were lymphoma cells, normal lymphocytes, myeloid leukemia cells, and normal and transformed fibroblasts, and the fibers were coated with different densities of concanavalin A or the lectins from soybean or wheat germ. Cells fixed with glutaraldehyde did not bind to lectin-coated fibers. The number of cells bound to fibers could be increased by increasing the density of lectin molecules on the fiber, the density of specific receptors on the cell, or the mobility of the receptors. It is suggested that binding of cells to fibers involves alignment and binding of specific cell surface receptors with lectin molecules immobilized on the fibers, and that this alignment requires short-range rapid lateral mobility (RLM) of the receptors. The titration of cell binding to fibers coated with different densities of lectin and albumin has been used to measure the relative RLM of unoccupied cell surface receptors for the lectin. The results indicate a relationship of RLM to lectin-induced cell-to-cell binding. The RLM or receptors for concanavalin A (Con A) was generally found to be higher than that of receptors for the lectins from wheat germ or soybean. Receptor RLM could be decreased by use of metabolic inhibitors or by lowering the temperature. Receptors for Con A had a lower RLM on normal fibroblasts than on SV40-transformed fibroblasts, and trypsinization of normal fibroblasts increased Con A receptor RLM. Normal lymphocytes, lymphoma cells, and lines of myeloid leukemia cells that can be induced to differentiate had a high receptor RLM, whereas lines of myeloid leukemia cells that could not be induced to differentiate had a low receptor RLM. These results suggest that the RLM of Con A receptors is related to the transformation of fibroblasts and the ability of myeloid leukemia cells to undergo differentiation

Topics: Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Dinitrophenols; Fibroblasts; Glycine max; Lectins; Leukemia, Myeloid; Lymphocytes; Lymphoma; Mice; Models, Biological; Nylons; Plant Lectins; Rats; Serum Albumin, Bovine; Simian virus 40; Temperature; Triticum

The mucoprotein, which is responsible for the formation of gastric mucous gel in pig, has been shown to bind equally well to suspensions of baby hamster kidney cells, polyoma-virus-transformed baby hamster kidney cells and HeLa cells. The binding of the mucoprotein to the cells is dependent on Ca 2

Topics: Animals; Azides; Binding Sites; Calcium; Cell Adhesion; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Clone Cells; Concanavalin A; Cricetinae; Gastric Mucins; HeLa Cells; Humans; Iodine Radioisotopes; Kidney; Magnesium; Mucoproteins; Pepsin A; Polyomavirus; Protein Binding; Serum Albumin, Bovine; Sulfur Radioisotopes; Swine; Temperature

Topics: Animals; Binding Sites, Antibody; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Hemadsorption; Humans; Kidney; Orthomyxoviridae; Polyomavirus

Treatment of untransformed mouse and hamster cells with the tertiary amine local anesthetics dibucaine, tetracaine and procaine increases their susceptibility to agglutination by low doses of the plant lectin concanavalin A. Agglutination of anesthetic-treated untransformed cells by low doses of concanavalin A is accompanied by redistribution of concanavalin A receptors on the cell surface to form patches, similar to that occurring in spontaneous agglutination of virus-transformed cells by concanavalin A. Immunofluorescence and freeze-fracture electronmicroscopic observations indicate that local anesthetics per se do not induce this redistribution of concanavalin A receptors but modify the plasma membrane so that receptor redistribution is facilitated on binding of concanavalin A to the cell surface. Fluorescence polarization measurements on the rotational freedom of the membrane-associated probe, diphenylhexatriene, indicate that local anesthetics produce a small increase in the fluidity of membrane lipids. Spontaneous agglutination of transformed cells by low doses of concanavalin A is inhibited by colchicine and vinblastine but these alkaloids have no effect on concanavalin A agglutination of anesthetic-treated cells. Evidence is presented which suggests that local anesthetics may impair membrane peripheral proteins sensitive to colchicine (microtubules) and cytochalasin-B (microfilaments). Combined treatment of untransformed 3T3 cells with colchicine and cytochalasin B mimics the effect of local anesthetics in enhancing susceptibility to agglutination by low doses of concanavalin A. A hypothesis is presented on the respective roles of colchicine-sensitive and cytochalasin B-sensitive peripheral membrane proteins in controlling the topographical distribution of lectin receptors on the cell surface.

Topics: Agglutination; Anesthetics, Local; Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Colchicine; Concanavalin A; Cricetinae; Cytochalasin B; Dibucaine; Dose-Response Relationship, Drug; Freeze Fracturing; Lectins; Mice; Mice, Inbred BALB C; Polarography; Polyomavirus; Procaine; Proteins; Simian virus 40; Temperature; Tetracaine

Topics: Agglutination Tests; Animals; Antigens, Neoplasm; Antigens, Viral; Cell Line; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Culture Techniques; Epithelial Cells; Fibroblasts; Fluorescent Antibody Technique; Kidney; Mice; Mice, Inbred Strains; Mutation; Polyomavirus; Virus Replication

Concanavalin A (Con A) has been popularly used as a cell surface probe for normal and neoplastic cells. Differences between normal fibroblasts and their transformed derivatives were examined using Con A agglutination, quantitative labeling with 125I-Con A and ultrastructural labeling with fluorescent- or ferritin-Con A. Con A agglutinates confluent-SV3T3 and 3T3 cells at midpoints of 20-60 and greater than, 500-2,000 mug/ml, respectively, and sparse cells at 5-15 and 1,200-1,500 mg/ml, respectively. Quantitative binding of 125I-Con A at 4 degrees C (10 or 15 min) with saturating lectin concentrations does not indicate a difference in the number of Con A receptors on sparse or confluent 3T3 and SV3T3 cells similar to many publications, but contrary to Noonan and Burger (1973). Under these conditions of labeling, ferritin-Con A is not internalized, indicating absence of endocytosis. The lateral mobility of Con A receptors and their relative ability to be aggregated on the cell surface by Con A was investigated with fluorescent- and ferritin-Con A. The initial distribution of Con A receptors on 3T3, SV3T3 and MSV3T3 cells under conditions of labeling at low temperature (0-5 degrees C) or to fixed cells was essentially randomly dispersed, but changes quickly to aggregated on SV3T3 and MSV3T3 (but not 3T3) after shifting the temperature to 20-37 degrees C, indicating, in general, a greater relative mobility of Con A receptors on SV3T3 and MSV3T3 cells. The aggregated Con A receptors seem to be directly involved in cell agglutination because they are usually found at the sites of cell-to-cell contact during 10 min agglutination experiments with ferritin-Con A. When confluent-3T3 cells are labeled on monolayer with ferritin-Con A at 0-4 degrees C, washed and then shifted to 20-37 degrees C for 10-15 min prior to in situ embedding, two classes of Con A receptors can be identified. One class appears to have low relative mobility and is associated with the 3T3 cell's extensive subplasma membrane microfilament network, while the other is capable of being aggregated and eventually endocytosed. On confluent-SV3T3 cells, only the latter class of receptors appears to be present, indicating a possible loss of cytoplasmic control over the distribution and mobility of lectin-binding sites on transformed cell surfaces.

Topics: Agglutination; Animals; Cell Transformation, Neoplastic; Cells; Cells, Cultured; Concanavalin A; Ferritins; Fibroblasts; Fluorescence; Mice; Mice, Inbred BALB C; Neoplasms, Experimental; Peptide Hydrolases; Receptors, Drug; Simian virus 40

We have studied the surface properties of four cell lines: normal human fibroblasts (GM-177), SV40 transformed Lesch-Nyhan fibroblasts (LN-SV), mouse macrophages, and somatic cell hybrids between mouse macrophages and human SV40-transformed fibroblasts (LN-SV). The concanavalin A (Con A)-peroxidase method was used to visualize the localization of the Con A binding sites by means of electron microscopy. The mean thickness of the reaction product was calculated and the distribution pattern was visualized by drawing "glycograms," from which the ratio of covered to uncovered areas was measured. Ruthenium red technique was used to study the ability of the cell surface to retain ruthenium red-positive materials after rinsing in phosphate-buffered saline. Before rinsing, there were no differences between thicknesses of the ruthenium red-positive surface layers of normal and virus-transformed Lesch-Nyhan fibroblasts. After rinsing, however, significant differences in the ability to retain ruthenium red-positive material were seen in favor of the SV40-transformed Lesch-Nyhan fibroblasts (54% over 35%). Before rinsing, significant differences in the thickness of the ruthenium red-positive material were found between mouse macrophages and the hybrids between mouse macrophages and SV40-transformed human fibroblasts, with the hybrids expressing a thicker ruthenium red-positive layer than did the parental mouse macrophages. After washing, however, the ability to retain this material was greater in mouse macrophages than in hybrids. The hybrid cell surface resembles that of SV40-transformed human Lesch-Nyhan fibroblasts. The Con A-peroxidase method shows a different distribution of the reaction product. In general, in all four cases the central portions of the cells had less continuous labeling patterns than did their peripheries, while the marginal portions were very often devoid of the reaction product. Normal human fibroblasts had the most continuous surface reaction product, the thickest reaction product, and the greatest Con A:gap ratio. In all four of these measurements, the hybrid cell line expressed great similarity to the parental cell lines, especially to the mouse macrophages.

Topics: Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Fibroblasts; Hybrid Cells; Macrophages; Peroxidases; Ruthenium Red; Simian virus 40

A transformed variant derived as a clone from normal 3T3 cells infected with simian virus 40 (SV40) has been found to possess a phenotype intermediate between that of normal cells and that characteristic of the transformed state, yet cells of the variant still test positively for the SV40-specific nuclear T-antigen. The variant exercises growth control, although not as stringently as do normal cells. Its cell size more closely resembles that of normal cells than of transformed cells. The variant also exhibits levels of spontaneous agglutination that are in line with those characteristic of the normal cells from which it was derived, and far higher than corresponding values for cells exhibiting the fully transformed phenotype. Plasma membranes of variant cells more closely resemble those of transformed cells than of normal cells as estimated by polyacrylamide gel electrophoresis. Perhaps the most distinguishing characteristic of the transformed variant is its complete immunity to agglutination by concanavalin A (Con A), even at concentrations of the lectin as high as 500 mug/ml. Moreover, trypsinization does not render variant cells as agglutinable in the presence of Con A as are untreated fully transformed cells. By contrast the variant displays a low tolerance of Con A toxicity, as monitored by ability to grow after treatment with the lectin, and on this count resembles transformed cells. Moreover a survey of several normal cell lines has revealed that even they do not consistently show resistance to Con A toxicity. These observations indicate that Con A-mediated agglutination and inability to grow after treatment with Con A are quite independent and do not bear a cause and effect relationship.

Topics: Agglutination; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Edetic Acid; Genetic Variation; Neoplasm Proteins; Phenotype; Proteins; Simian virus 40; Trypsin

Lymphocytes from normal adults, with or without serological signs of previous Epstein-Barr virus (EBV) infection, could be stimulated to proliferate and produce killer cells by incubation with autologous EBV-genome-positive lymphoid cell lines (LCLs). The stimulated cells were most probably of T-cell origin, although at the peak of stimulation many of them lacked the sheep erythrocyte marker. Direct effector-target cell contact was necessary for lysis to occur. The cytotoxicity of autologously stimulated (AS) lymphocytes was not restricted to EBV-genome-positive LCLs, nor to cell lines of hematopoietic origin. It was equally broad if cells carrying complement receptor had been removed before stimulation. Fresh lymphocytes, blasts induced by phytohemagglutinin or concanavalin A, and Burkitt's lymphoma biopsy cells were resistant or considerably less sensitive. Mouse cells--even cell lines--were resistant. The sensitivity of target cells to lysis correlated positively with their capacity to block AS lymphocyte lysis of autologous LCLs in competition experiments. The cytotoxicity of AS lymphocytes was blocked by EBV-genome-positive and -negative cell lines, whereas the EBV-related cytotoxicity of T cells from acute cases of infectious mononucleosis was blocked by EBV-genome-positive LCL only.

Topics: Animals; Antigen-Antibody Reactions; Burkitt Lymphoma; Cell Line; Cell Transformation, Neoplastic; Complement System Proteins; Concanavalin A; Cytotoxicity Tests, Immunologic; Herpesvirus 4, Human; Humans; Lectins; Leukemia; Lymphocyte Activation; Lymphocyte Transfusion; Lymphocytes; Lymphoma; Mitomycins; Stimulation, Chemical; T-Lymphocytes; Transplantation, Autologous

The infectivity of avian RNA tumor viruses was inactivated to varying degrees by treatment with either concanavalin A (Con A) or phytohemagglutinin but not by treatment with wheat germ agglutinin. In general, leukosis viruses reacted preferentially with Con A, whereas sarcoma viruses showed more affinity for phytohemagglutinin. In a more extensive study with subgroup A of Prague Rous sarcoma virus (PR-A), the effect of inactivation by Con A could be specifically prevented by the addition of alpha-methyl-D-mannoside, alpha-methyl-D-glucoside, and N-acetyl-D-glucosamine. These sugars were also capable of eluting [3H]glucosamine-labeled material from disrupted PR-A virus, which was bound to a Con A-sepharose affinity column. A major viral glycoprotein recovered from the column had the same mobility as gp85 in polyacrylamide gel electrophoresis and could be immunoprecipitated with anti-gp85 antiserum. These results suggest that the material reacting with Con A is present on the gp85 component of the viral glycoprotein. The diversity in the reactivity of the glycoproteins of transforming and nontransforming viruses with plant lectins is discussed.

Topics: Acetylgalactosamine; Acetylglucosamine; Alpharetrovirus; Avian Leukosis Virus; Avian Sarcoma Viruses; Binding Sites, Antibody; Cell Transformation, Neoplastic; Concanavalin A; Culture Techniques; Genetic Variation; Glycoproteins; Lectins; Methylglucosides; Methylmannosides; Plant Lectins; Triticum; Vegetables; Viral Proteins; Virus Replication

The observation of BOREK et al. (1973) on nonagglutinability of transformed rat liver cells by Lens culinaris lectin and our ultrastructural findings of a greater mobility of the Lens culinaris lectin receptors on transformed rat liver cells as compared to normal rat liver cells (ROTH 1975) initiated the present agglutination experiments on liver cells with lectins. For agglutination assay the microhemadsorption technique after FURMANSKI et al. (1973) was used with exception of several tests on EDTA-detached cells. The transformed rat liver cells exhibited, in contrast to the findings of BOREK et al. (1973), a positive microhemadsorption with Lens culinaris lectin as well as with Concanavalin A, Ricinus communis lectin and wheat germ agglutinin whereas the normal rat liver cells became positive only after a brief trypsin treatment. The significance of the difference in agglutinability of rat liver cells with Lens culinaris lectin and the other lectins used is discussed with regard to the cell-cell interaction mediated by lectins.

Topics: Agglutination Tests; Animals; Binding Sites; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; Cell Wall; Concanavalin A; Hemadsorption; Lectins; Liver; Liver Neoplasms; Plant Lectins; Plants; Plants, Toxic; Rats; Ricinus; Surface Properties; Triticum

Cultures of human adenoidal lymphocytes exposed briefly to either phytohemagglutinin (PHA), Staphylococcus filtrate (Staph-F), concanavalin-A (Con-A), or pokeweed mitogen (PWM) incorporate increased amounts of thymidine earlier than replicate cultures exposed continuously to the mitogens. These effects can begin in the first 24 hr of culture and are seen maximally between 36 and 72 hr. Once a blastogenic response is established, PHA or PWM can diminish that response. Inhibition with PWM requires that the initial stimulation was with this mitogen, while PHA can inhibit blastogenesis to both PHA and PWM-stimulated cells. Because these mitogens can have a paradoxical effect on adenoidal lymphocytes, being capable of both initiating and inhibiting DNA synthesis, this phenomena should be kept in mind when such systems are utilized for the evaluation of antigens and drug effects.

Topics: Adenoids; Cell Transformation, Neoplastic; Concanavalin A; Depression, Chemical; DNA; Humans; Kinetics; Lectins; Lymphocytes; Mitogens; Staphylococcus; Thymidine; Tritium

Subclones isolated from a Chinese hamster hybrid line, derived from fusion of an actinomycin D-resistant and an actinomycin D-sensitive strain, were studied with respect to their resistance to actinomycin D, karyology, transplantability and agglutination by concanavalin A. Statistical analysis of the results allowed the establishment of a classification of the strains based on increasing resistance to actinomycin D. There appeared to be an inverse correlation between actinomycin D-resistance and tumorigenicity and a positive correlation between this resistance and the presence of a marker chromosome.

Topics: Agglutination Tests; Animals; Cell Transformation, Neoplastic; Chromosomes; Clone Cells; Concanavalin A; Cricetinae; Dactinomycin; Hybrid Cells; Hybridization, Genetic; Immunity, Cellular; Karyotyping; Neoplasm Transplantation; Neoplasms, Experimental

Wheat germ agglutinin and Concanavalin A were found to agglutinate a variety of normal hamster cells as well as a number of lines of transformed or tumor cells. The normal cells included preparations from embryos and neonatal organs, and the spontaneous line, BHK-21 C13. In order to explore more fully the situation in regard to the BHK cells, three sublines were isolated and studied. A "flat revertant" selected by FUdR treatment grew to diminished saturation density and failed to produce colonies in soft agar in contrast to the parent line, which did. Yet, it was only minimally less tumorigenic and less agglutinable than the parent line. Lines derived from one and four consecutive in vivo tumor passages showed increased tumorigenicity and growth in soft agar when compared to the parent line. Although the first passage line showed increased saturation density, the saturation density of the fourth passage line was similar to or less than that of the parent cells. Agglutinability of these lines was not significantly increased. These results indicate that, in contrast to widely accepted beliefs, tumorigenicity may correlate poorly with growth characteristics and agglutination by plant lectins.

Topics: Agglutination; Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Lectins; Neoplasm Transplantation; Neoplasms, Experimental; Phenotype; Transplantation, Homologous

Topics: Agglutination; Amino Acid Sequence; Biological Transport; Carbohydrates; Cell Transformation, Neoplastic; Chemical Phenomena; Chemistry; Concanavalin A; Hydrolysis; Protein Binding; Surface Properties; Trypsin

The ability of suspensions of BALB/c cells to catalyze the incorporation of nucleotide sugars into complex polysaccharides has been compared. These cells have previously been characterized for concanavalin A-induced agglutinability, tumorigenicity, and malignancy. All of the cell lines tested catalyze transfer of the sugar moieties of cytosine 5-monophosphate N-acetylneuraminic acid, uridine 5-diphosphate galactose, uridine 5-diphosphate N-acetylgalactosamine, uridine 5-diphosphate N-acetylglucosamine, uridine 5-diphosphate glucose, and guanidine 5-diphosphate monnose into glycoproteins and glycolipids. While some transformed lines exhibit alterations in transferase levels, others cannot be distinguished from normal cells. Normal cells, transformed cells that cause tumors that regress, and transformed cells that cause tumors that kill an immunologically competent host show growth-dependent changes in transferase activities. Determining the ability to catalyze carbohydrate transfer is insufficient for predicting the tumorigenic and malignant properties of a cell line.

Topics: Agglutination; Animals; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cytosine; Galactosamine; Galactose; Glucosamine; Glycolipids; Glycoproteins; Glycosides; Guanidines; Hexosyltransferases; Mannose; Mice; Mice, Inbred BALB C; Neoplasms, Experimental; Neuraminic Acids; Nucleoside Diphosphate Sugars; Nucleotides; Uridine Diphosphate Sugars

Topics: Agglutination; Anesthetics, Local; Animals; Binding Sites; Calcium; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Dibucaine; Fibroblasts; Mice; Mice, Inbred BALB C; Microscopy, Electron; Procaine; Tetracaine

Well resolved 1-H and 13-C NMR spectra were obtained with normal and SV 40-transformed cell membranes. Estimation of the ratio of 13-CT2 values of the normal to transformed cell membranes showed an increased intermolecular motion in the transformed cell membranes. The temperature dependence of the (CH2) line in the 1-H spectra in the temperature range 298-343 degrees K shows an activation energy for the lateral diffusion of the fluid phospholipid regions in the normal cell membranes while the transformed ones show practically no temperature dependence in this temperature range. The fluidity of the phospholipid region in the transformed cell membrane seems to be significantly higher than that observed in the normal cell material. These data support and extend the findings concerning the mobility of the concanavalin A binding/agglutinating sites on the surface of normal and virus-transformed cells and suggest further approaches to the study of the membrane alterations in tumor cells.

Topics: Animals; Carbon Isotopes; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Embryo, Mammalian; Fibroblasts; Hydrogen; Lipids; Magnetic Resonance Spectroscopy; Mathematics; Molecular Conformation; Phospholipids; Receptors, Drug; Temperature

Cultured normal and transformed fibroblasts were treated "in situ" by the concanavalin A-peroxidase labelling technique. It is known that peroxidase recognizes only a fraction of the bound lectin depending on the cell type. Kinetics studies revealed that 80 to 95 percent of the peroxidase and only 10 percent of the lectin are released from the cell surface when the labelled cells were reincubated at 37 degrees C. It is shown that it is mostly the concanavalin traced by peroxidase that is released and also that the lectin and the enzyme are shed as a complex or concomitantly. Consequently, the shedding pattern of the enzyme is used to demonstrate heterogeneity in the lectin binding sites; there are two main components labelled by concanavalin and peroxidase, one which has a short period (from 6 to 16 min) and another one with a much longer one (1.3 to 3 h). It is shown that when cells are incubated at 37 degrees C after a lectin treatment, secondary binding forces occur between the lectin and cell surface components which render the lectin unavailable for inhibiting sugars. Under the same conditions, some peroxidase can still be bound and a slight agglutination can still occur.

Topics: Animals; Binding Sites; Binding, Competitive; Cell Adhesion; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Fibroblasts; Glucose; Mannose; Methylmannosides; Peroxidases; Time Factors

Calculations of the density of Concanavalin A (Con A)-binding sites on normal and transformed fibroblasts have, as yet, been based on the unproven assumption that suspended cells are smooth spheres. We studied the surface morphology of suspended normal and transformed fibroblasts with scanning and transmission electron microscopes, and found a large difference in surface morphology between suspended normal and transformed 3T3 cells. When this difference in surface morphology was taken into account, the estimated cell surface area of normal 3T3 cells was approximately seven times larger than that of transformed 3T3 cells. Since equal numbers of 3H-Con A molecules are bound on normal and transformed cells, the density of Con A-binding sites is approximately seven times greater on transformed than on normal 3T3 cells. The difference in density of Con A-binding sites between normal and transformed fibroblasts might be sufficient to explain the difference in agglutination response, as originally suggested by Burger, and may also be the cause of the different degrees of clustering of Con A-binding sites on the plasma membrane of these cells.

Topics: Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Methylglucosides; Surface Properties

Topics: Agglutination; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Lectins; Receptors, Drug

Topics: Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Histocytochemistry; Lectins; Mitosis; Organoids; Pinocytosis; Receptors, Drug

Topics: Adenosine Triphosphate; Agglutination; Binding Sites; Cell Count; Cell Division; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; DNA; Trypsin

Comparative electron microscopic investigations were performed in living cultures of normal rat liver cells, of rat liver cells in vivo transformed by diethylnitrosamine and on Zajdela ascites hepatoma cells of the rat concerning the mobility of the Concanavalin A cell surface receptors. The cells were incubated in Concanavalin A and peroxidase and subsequently washed. They were then reincubated for various periods at +37 degrees C in PBS prior to fixation. In the case of the Zajdela ascites hepatoma cells the cells were reincubated after Concanavalin A incubation followed by fixation and peroxidase incubation. The cytochemical procedure allowed us to show differences in the mobility of Concanavalin A surface receptors between normal and transformed rat liver cells. The cell surface label disappeared completely within 15 min of reincubation in the transformed cells, whereas in normal cells the same degree of loss in surface label was visible after 120 min reincubation. In both cases an internalization of labelled plasma membrane areas occurred. After complete disappearance of cell surface label in diethylnitrosamine transformed cells a complete relabelling of the cell surface occurred after 60 min reincubation caused by an exocytosis.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Diethylnitrosamine; Liver Neoplasms; Microscopy, Electron; Neoplasms, Experimental; Rats; Receptors, Drug

Topics: Adenoviridae; Agglutination; Animals; Antigens, Neoplasm; Antigens, Viral; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chromosomes; Concanavalin A; Cricetinae; Genetic Variation; Glycoproteins; Kidney; Neoplasm Proteins; Sialic Acids

Topics: Adenosine Triphosphate; Agglutination; Azides; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Dinitrophenols; Glucose; Glycine max; Iodoacetates; Kinetics; Lectins; Oligomycins; Plant Lectins; Receptors, Drug; Temperature

The distribution of surface-bound concanavalin A on the membranes of 3T3, and simian virus 40-transformed 3T3 cultured mouse fibroblasts was examined using a shadow-cast replica technique with a hemocyanin marker. When cells were prefixed in paraformaldehyde, the binding site distribution was always random on both cell types. On the other hand, labeling of transformed cells with concanavalin A (Con A) and hemocyanin at 37 degrees C resulted in the organization of Con A binding sites (CABS) into clusters (primary organization) which were not present on the pseudopodia and other peripheral areas of the membrane (secondary organization). Treatment of transformed cells with colchicine, cytochalasin B, or 2-deoxyglucose did not alter the inherent random distribution of binding sites as determined by fixation before labeling. However, these drugs produced marked changes in the secondary (but not the primary) organization of CABS on transformed cells labeled at 37 degrees C. Colchicine treatment resulted in the formation of a caplike aggregation of binding site clusters near the center of the cell, whereas cytochalasin B and 2-deoxyglucose led to the formation of patches of CABS over the entire membrane, eliminating the inward displacement of patches observed on untreated cells. The distribution of bound Con A on normal cells (3T3) at 37 degrees C was always random, in both control and drug-treated preparations. Pretreatment of cells with Con A enhanced the effect of colchicine on cell morphology, but inhibited the morphological effects of cytochalasin B. The mechanisms that determine receptor movement and disposition are discussed.

Topics: Agglutination; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Cytochalasin B; Fibroblasts; Glucose; Hemocyanins; Iodine Radioisotopes; Mice; Mice, Inbred BALB C; Microscopy, Electron; Microscopy, Phase-Contrast; Pseudopodia; Simian virus 40; Staining and Labeling; Vinblastine

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chromosomes; Colchicine; Concanavalin A; Fibroblasts; Floxuridine; Gammaretrovirus; Genetic Variation; Idoxuridine; Mice; Mice, Inbred Strains; Mutation; RNA-Directed DNA Polymerase; Sarcoma

The distribution of Concanavalin A receptors in cultures of normal rat liver cells and of Zajdela ascites hepatoma cells of the rat was investigated by the Con A-po technique (Bernhard and Avrameas, 1971). Con A-po labelling of the cell surface showed a patchy distribution of Concanavalin A receptors in ascites hepatoma cells, whereas continuous Con A-po labelling was seen at the surface of normal rat liver cells. The different distribution of Concanavalin A receptors was caused by rapid internalization of Con A-po labelled material and not by a simple rearrangement of the receptors. The significance of this process is discussed with respect to Concanavalin A receptor mobility and membrane stability after cell transformation.

Topics: Animals; Carcinoma, Hepatocellular; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Culture Media; Golgi Apparatus; Liver; Liver Neoplasms; Microscopy, Electron; Neoplasm Transplantation; Neoplasms, Experimental; p-Dimethylaminoazobenzene; Peroxidases; Rats; Receptors, Drug; Staining and Labeling

Mouse neuroblastoma cells (clone neuro-2A) in the undifferentiated and "differentiated" form were compared by light and electron microscopy. "Cytodifferentiation" was induced in monolayer cultures by the addition of dibutyryl-cyclic AMP. The pattern of concanavalin A binding sites was studied after coupling with horseradish peroxidase. The following major differences were observed. The differentiated cells are characterized by numerous and long neurites, aggregation of ribosomes into polysomes, an extensive network of neurofilaments and microtubules, many dense-core neurosecretory-like vesicles, a discontinuous pattern of concanavalin A binding sites on the plasma membrane, and an increase of the specific activities of acetylcholinesterase, choline acetylase and tyrosine hydroxylase. In contrast, the undifferentiated cells grown in suspension culture lack neurites, contain dispersed ribosomes, infrequent neurofilaments and microtubules and dense-core neurosecretory-like vesicles, and exhibit a continuous pattern of concanavalin A binding sites. In addition, the specific activities of the above mentioned enzymes are significantly lower.

Topics: Acetylcholinesterase; Acetyltransferases; Animals; Binding Sites; Bucladesine; Cell Line; Cell Membrane; Cell Nucleus; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Mice; Mice, Inbred A; Microscopy, Electron; Microtubules; Neoplasms, Experimental; Neuroblastoma; Organoids; Peroxidases; Polyribosomes; Ribosomes; Tyrosine 3-Monooxygenase

Topics: Adult; Agglutination; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Depression, Chemical; Fibroblasts; Glioma; Glucose; Glutaral; Humans; Hyaluronoglucosaminidase; In Vitro Techniques; Lactose; Lectins; Lung; Methods; Neuroglia; Plant Lectins; Plants, Toxic; Ricinus; Sarcoma; Stimulation, Chemical; Temperature; Trypsin

Topics: Animals; Antigen-Antibody Reactions; Antigens, Viral; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Concanavalin A; Cytotoxicity Tests, Immunologic; Fibroblasts; Immunization; Lectins; Lymph Nodes; Lymphocytes; Mice; Mice, Inbred BALB C; Simian virus 40; Spleen

Topics: Adenoviridae; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Embryo, Mammalian; Histocytochemistry; Humans; Lectins; Microscopy, Electron; Peroxidases; Polyomavirus; Protein Binding; Rats; Simian virus 40

Topics: Acetates; Animals; Carbohydrates; Carbon Radioisotopes; Cell Adhesion; Cell Fractionation; Cell Transformation, Neoplastic; Cetylpyridinium; Chromatography, Gel; Chromatography, Ion Exchange; Chromatography, Paper; Clone Cells; Concanavalin A; Fibroblasts; Glycols; Glycoproteins; Glycosaminoglycans; Hyaluronoglucosaminidase; Mice; Mice, Inbred BALB C; Molecular Weight; Simian virus 40; Solubility; Sulfuric Acids; Tritium

Topics: Adenoviridae; Agglutinins; Aldehydes; Animals; Cell Line; Cell Membrane Permeability; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Dinitrophenols; Iron Chelating Agents; Mannose; Microscopy, Electron; Nickel; Peroxidases; Polyomavirus; Radioisotopes; Simian virus 40; Surface Properties; Time Factors

Topics: Agglutination; Animals; Cell Aggregation; Cell Count; Cell Transformation, Neoplastic; Clone Cells; Colchicine; Concanavalin A; Cricetinae; Depression, Chemical; Fibroblasts; Kidney; Kinetics; Polyomavirus; Vinblastine

Non-virus-producing NIH/3T3 cells transformed by the murine sarcoma virus are agglutinated by conconavalin A to the same low level as normal NIH/3T3 cells. Infection with the murine leukemia virus greatly increases the agglutination of transformed cells but not that of normal cells. These data suggest that the morphological expression of cell transformation and the surface alterations associated with increased cell agglutination are controlled by the expressions of different sarcoma virus genes.

Topics: Agglutination; Animals; Cell Line; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Fibroblasts; Gammaretrovirus; Genotype; Leukemia Virus, Murine; Mice; Mice, Inbred Strains; Sarcoma

Topics: Agglutination; Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Contact Inhibition; Graft Rejection; Male; Mice; Mice, Inbred BALB C; Neoplasms; Simian virus 40

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Binding Sites, Antibody; Carcinogens; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Edetic Acid; Fibroblasts; Fluoresceins; Lectins; Mice; Mitosis; Nitrosamines; Polyomavirus; Simian virus 40; Tritium; Trypsin

Topics: Animals; Binding Sites; Brain; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Hybrid Cells; Indicators and Reagents; Mice; Peroxidases; Rats; Receptors, Drug; Simian virus 40

Topics: Agglutination Tests; Animals; Arginine; Cell Division; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chlorine; Concanavalin A; Contact Inhibition; Fibroblasts; Lysine; Mice; Protease Inhibitors; Simian virus 40; Tosyl Compounds

Topics: Agglutination; Animals; Autoradiography; Avian Sarcoma Viruses; Bromodeoxyuridine; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chickens; Concanavalin A; Genotype; Kidney; Neoplasm Transplantation; Sarcoma, Avian; Thymidine; Transplantation, Heterologous; Transplantation, Homologous; Tritium; Virus Replication

Topics: Azides; Bucladesine; Cell Line; Cell Survival; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Cyanides; Female; Fluorides; Glycolysis; Iodine Radioisotopes; Iodoacetates; Kinetics; Ovary; Oxidative Phosphorylation; Receptors, Drug; Testolactone; Time Factors

Topics: Animals; Binding Sites; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; Chromatography, Gas; Chromatography, Gel; Concanavalin A; Glycopeptides; Haptens; Lectins; Liver; Liver Neoplasms; Models, Biological; Neoplasms, Experimental; Rats; Receptors, Drug; Spectrophotometry; Spectrophotometry, Ultraviolet; Trypsin

Topics: Agar; Agglutination Tests; Animals; Bromodeoxyuridine; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chromatography, Paper; Concanavalin A; Mice; Neoplasms, Experimental; Polyomavirus

Topics: Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; Electrophoresis, Disc; Erythrocytes; Humans; In Vitro Techniques; Iodine Radioisotopes; Methods; Mice; Mice, Inbred BALB C; Simian virus 40

Topics: Agglutination Tests; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Dogs; Ferritins; Iodine Radioisotopes; Kidney; Lectins; Microscopy, Electron; Orthomyxoviridae; Plant Lectins; Plants, Toxic; Polyomavirus; Protein Biosynthesis; Ricinus; Time Factors; Vesicular stomatitis Indiana virus; Virus Cultivation

Revertants of Kirsten sarcoma virus transformed nonproducer BALB/3T3 cells (KA31 cells) were isolated after exposing the transformed cells to 5-fluorodeoxyuridine at high cell density, or when suspended in methylcellulose. Revertants were also isolated by treating KA31 cells with the lectin, concanavalin A, which is manyfold more toxic to transformed cells than for normal cells. The revertants resemble BALB/3T3 cells in their morphology and growth characteristics in that they have a low saturation density, fail to grow in 1% calf serum or when suspended in methylcellulose, and cease to synthesize DNA after reaching their saturation density. Infection by murine leukemia virus rescues Kirsten sarcoma virus from only the concanavalin-A-selected variants, though all the revertants are susceptible to infection by leukemia virus. The concanavalin A revertants also become transformed after infection with murine leukemia virus. All the revertants can be transformed by Kirsten sarcoma virus but not by simian virus 40.

Topics: Animals; Autoradiography; Cell Transformation, Neoplastic; Cells, Cultured; Chromosomes; Concanavalin A; DNA; DNA, Neoplasm; Floxuridine; Gammaretrovirus; Helper Viruses; Leukemia Virus, Murine; Methylcellulose; Mice; Mice, Inbred BALB C; Simian virus 40; Thymidine; Tritium; Viral Plaque Assay; Virus Replication

Topics: Agglutination; Animals; Binding Sites, Antibody; Binding, Competitive; Carbon Radioisotopes; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Contact Inhibition; Fibroblasts; Iodine Radioisotopes; Mice; Polyomavirus; Simian virus 40; Succinates; Thymidine; Tritium

Topics: Agglutination Tests; Animals; Binding Sites; Binding, Competitive; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Chromatography, Affinity; Chromatography, Gel; Computers; Concanavalin A; Cricetinae; Electrophoresis, Polyacrylamide Gel; Kinetics; Macromolecular Substances; Microscopy, Electron; Nickel; Peroxidases; Protein Binding; Protein Conformation; Radioisotopes; Receptors, Drug; Simian virus 40; Surface Properties; Temperature; Ultracentrifugation

We have devised a quantitative way to measure the agglutination of cells which utilizes the size discrimination feature of an automatic particle counter. With this method we have studied the agglutinability by concanavalin A of 3T3 cells, a mutant of 3T3 cells (3T3cAMP(tcs)) in which cyclic AMP levels fall when the cells are subjected to temperature change or fresh serum, and L929 cells. We find with 3T3cAMP(tcs) cells that low levels of cyclic AMP correlate with increased agglutinability and that high levels of cyclic AMP correlate with decreased agglutinability. Prior treatment of these cells with a cyclic AMP phosphodiesterase inhibitor or Bt(2)cAMP blocks the increase in agglutinability induced by temperature change. When 3T3 cells are treated with fresh serum, their agglutinability also increases although to a much smaller extent than with 3T3cAMP(tcs) cells. Cells change their agglutinability very rapidly. Treatment of L929 cells for 15 min with 1-methyl-3-isobutyl xanthine at 1 mM decreases their agglutinability to the level of normal 3T3 cells. We conclude that in normal and transformed cells the level of cyclic AMP regulates agglutinability.

Topics: Agglutination; Agglutination Tests; Animals; Bucladesine; Cell Count; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fibroblasts; L Cells; Mice; Temperature

Topics: Agglutination; Agglutination Tests; Animals; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Centrifugation, Density Gradient; Concanavalin A; Culture Techniques; Fibroblasts; Hemadsorption; Mice; Simian virus 40; Surface Properties

Topics: Agglutination; Agglutination Tests; Animals; Avian Sarcoma Viruses; Bromodeoxyuridine; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Fibroblasts; Glucose; Mutation; Temperature; Thymidine; Tritium

Numerous studies have reported the capacity of the lectin, concanavalin A, to agglutinate selected cell-types. The finding that cells transformed in culture, embryonic cells, and malignant cells are all agglutinated by this substance, may contribute to our understanding of the oncogenic process. The present study compared the response to concanavalin A of rat hepatocytes derived from livers of differing developmental and mitotic-status as well as those derived from malignant liver tumors (hepatomas). Fetal hepatocytes and hepatoma cells were highly susceptible to agglutination while hepatocytes from post-natal livers, whether dividing or quiescent, were not. Treatment with protease(s) did not make the interphase hepatocyte agglutinable. These data emphasize the importance of examining a wide variety of cells in attempting to understand the interaction of lectins on cell surfaces, and further, demonstrate the value of obtaining cells directly from tissue(s) during differing physiologic and pathologic states.

Topics: Agglutination; Animals; Carcinoma, Hepatocellular; Cell Division; Cell Separation; Cell Transformation, Neoplastic; Concanavalin A; Erythrocytes; Liver; Liver Neoplasms; Male; Microbial Collagenase; Rats; Rats, Inbred ACI; Trypsin

Topics: Agglutination Tests; Animals; Binding Sites; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Edetic Acid; Erythrocytes; Ferritins; Fluoresceins; Hemagglutination Tests; Lectins; Mice; Microscopy, Electron; Peroxidases; Polyomavirus; Time Factors

Topics: Agglutination Tests; Animals; Binding Sites; Cell Aggregation; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chromatography, DEAE-Cellulose; Concanavalin A; Edetic Acid; Electrophoresis, Disc; Endocytosis; Kinetics; Methods; Methylglycosides; Mice; Receptors, Drug; Simian virus 40; Solubility; Time Factors; Tritium; Trypsin

A substance capable of promoting tumour cell aggregation was released from rat ascites hepatoma cell (possibly from the cell surface) kept in Hanks' balanced salt solution (free of calcium and magnesium) in the cold, and then partially purified by chromatography with DEAE-Sephadex and gel filtration with Bio-gel. The thermostable substance seemed to be a glycoprotein and its molecular weight was about 72,000 when measured by gel filtration on Sephadex G-200. It had no proteolytic activity. The material was clearly effective for rat ascites hepatoma cells as well as SV40 transformed cells, but less effective for Chang's cells and apparently ineffective for normal rat liver cells and red blood cells. The action of this material was more potent than that of Jack bean concanavalin A when assayed for aggregation of SV40 transformed cells. Its effect was not influenced by concanavalin A inhibitors such as alpha-methyl-D-glucopyranoside, N-acetyl-D-glucosamine and D-glucose.

Topics: Animals; Carcinoma, Hepatocellular; Cell Aggregation; Cell Separation; Cell Transformation, Neoplastic; Chromatography, Gel; Concanavalin A; Erythrocytes; Glucosamine; Glucose; Glycoproteins; Liver; Liver Neoplasms; Male; Methylglucosides; Molecular Weight; Rats; Simian virus 40; Tissue Extracts

Topics: Agglutination; Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Culture Techniques; Electricity; Ferritins; Fibroblasts; Lymphocytes; Lymphoma; Lysine; Models, Biological; Moloney murine leukemia virus; Neoplasms, Experimental; Neuraminidase; Polymers; Rats; Simian virus 40; Surface Properties

Tumors induced by SV40 virus in newborn and adult Syrian hamsters were analyzed for their growth properties in vitro, transplantation, agglutination with concanavalin A and chromosome constitution. Following transfer to culture flasks tumor cells grew for 2-3 weeks, then went through a "degenerative phase". Eventually lines were established which became adapted to in vitro culture conditions. Agglutination properties of these tumor cells were similar to those of cells transformed in vitro with SV40 virus. Of five subcutaneous and six intratesticular tumors that were analyzed for their chromosome constitution, all had a hypotetraploid chromosome number distribution. The results of the chromosome studies suggest a similiar pattern in the evolution of malignant properties of cells transformed in vitro and in vivo following infection with SV40 virus.

Topics: Agglutination; Animals; Antigens, Neoplasm; Antigens, Viral; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chromosome Aberrations; Chromosomes; Concanavalin A; Cricetinae; Diploidy; Fibrosarcoma; Karyotyping; Male; Neoplasm Transplantation; Neoplasms, Experimental; Polyploidy; Simian virus 40; Testicular Neoplasms; Transplantation, Homologous

Topics: Agglutination Tests; Animals; Cell Survival; Cell Transformation, Neoplastic; Concanavalin A; Culture Techniques; Kidney; Kidney Neoplasms; Mitosis; Nitrosamines; Platelet Adhesiveness; Rats

When BALB/c 3T3, simian virus 40 (SV40)-transformed 3T3 (SVT2), and revertant variants of the transformed cells are removed by EGTA treatment from the substrate on which they were grown, they leave behind a layer of glycoprotein which has been characterized biochemically (Terry, A. H. and L. A. Culp. 1974. Biochemistry. 13:414.)-substrate-attached material (SAM). The influence of SAM from normal and from transformed cells on cellular attachment to the substrate, morphology, movement, and growth has been examined. All three cell types displayed a 30% higher plating efficiency when grown on 3T3 SAM. The morphology of SVT2 colonies and of individual SVT2 cells was dramatically affected by growth on 3T3 SAM-the cells (a) were more highly spread on the substrate, (b) resisted crawling over neighboring cells, and (c) resisted movement away from the edge of colonies; SVT2 SAM was not effective in causing these changes. A cell-to-substrate attachment assay using thymidine-radiolabeled cells and untreated or SAM-coated cover slips was developed. SVT2 cells attached to 3T3 SAM- or SVT2 SAM-coated cover slips with a faster initial rate and to a higher saturation level than to untreated substrate, whereas 3T3 and revertant cells exhibited no preference; there was no species specificity in these cell-substrate attachment phenomena. Trypsin-released cells attached much more slowly than EGTA-released cells. 3T3 SAM, however, was not effective in lowering the saturation density of mass cultures of virus-transformed cells. These experiments suggest that the substrate-attached glycoproteins of normal cells affect the cellular adhesivity, morphology, movement, and perhaps growth patterns of virus-transformed cells-i.e., causing partial reversion of these properties of transformed cells to those found in contact-inhibited fibroblasts. A model for the involvement of substrate-attached glycoproteins in cell-to-substrate adhesion, and possibly cell-to-cell adhesion, has been proposed.

Topics: Animals; Cell Adhesion; Cell Division; Cell Movement; Cell Transformation, Neoplastic; Cells, Cultured; Chelating Agents; Concanavalin A; Contact Inhibition; Ethylene Glycols; Fibroblasts; Genetic Variation; Glycoproteins; Mice; Models, Biological; Neoplasm Proteins; Thymidine; Tritium; Trypsin

Topics: Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fibroblasts; Molecular Weight; Nickel; Peroxidases; Plant Lectins; Plants; Protein Binding; Radioisotopes; Receptors, Drug; Spectrophotometry

Topics: Agglutination; Animals; Binding Sites, Antibody; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fibroblasts; Lectins; Mice; Mitosis; Surface Properties

The growth of 3T3 and SV101 3T3 cells in a lipid-depleted medium is enhanced by the addition of biotin or some fatty acids. The extent of enhancement depends on the fatty acid(s) supplied. The presence of linoleate is unique, since it induces a morphological alteration in 3T3 cells resulting in a cell similar to an SV101-transformed 3T3 cell. Analyses of the fatty acids from the membrane phosphatides show that the exogenously supplied fatty acids are incorporated and alter the fatty acid composition. This is most clearly evident with heptadecanoate-grown cells, in which this fatty acid and its derivatives comprise over 45% of the fatty acids in the phospholipids. The fatty acid replacements have a striking effect on the temperature dependence of agglutination by wheat germ agglutinin and concanavalin A, implying that fluidity is involved in agglutination. These temperature dependencies and the effect of fatty acid replacements on them were different for the two lectins, but similar for both transformed and untransformed cells. These observations are interpreted as suggesting that the lipid phase is heterogeneous, and that transformed and untransformed cell membranes have regions of similar fluidity.

Topics: Agglutination; Animals; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fatty Acids, Essential; Fibroblasts; Lectins; Mice; Temperature

The cell-cell binding induced by concanavalin A between single cells has been analyzed by use of cells attached to nylon fibers. Binding of a concanavalin A-coated cell to an untreated cell was found to a high degree between two lymphoma tumor cells, less frequently between a lymphoma cell and a normal lymphocyte, and only rarely between two normal lymphocytes. The binding was inhibited by the presence of a saccharide inhibitor of concanavalin A, but could not be reversed by addition of the inhibitor after the cells had bound to each other. Although no binding was obtained when both cells were coated with lectin or fixed with glutaraldehyde, fixation of a cell before coating with concanavalin A enhanced its ability to bind an untreated cell. The results indicate that cell-cell binding induced by concanavalin A requires short-range lateral movement of cell receptors for the lectin, that only one cell has to have mobile receptors, and that some receptors must be unoccupied by lectin molecules before cell-cell contact. Clustering of the receptors is not necessary and seems to hinder cell-cell binding. It is suggested that the short-range movement is required for alignment of individual receptors so as to form multi-point bridges between two cells by lectin molecules. The bridging is then followed by the formation of irreversible bonds between the cells. The receptors on tumor cells appear to have a greater ability than receptors on normal cells to align themselves for cell-cell binding.

Topics: Agglutination; Animals; Antibodies, Neoplasm; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Leukemia, Experimental; Lymphocyte Activation; Lymphocytes; Lymphoma; Mice; Mice, Inbred A; Microscopy, Fluorescence; Moloney murine leukemia virus; Rats; T-Lymphocytes

Topics: Agglutination; Agglutination Tests; Animals; Binding Sites, Antibody; Carbohydrates; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fibroblasts; Fluorescent Antibody Technique; Glycogen; Leukemia, Experimental; Leukemia, Myeloid, Acute; Lymphocytes; Lymphoma; Male; Mice; Movement; Rats; Tritium; Trypsin

Topics: Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Leukocytes; Lymphocytes; Phagocytosis; Protein Binding; Surface Properties

Topics: Benzopyrenes; Blood; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; L Cells; Lectins; Methylcellulose

Topics: Animals; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Fluoresceins; Lectins; Methods; Mice; Simian virus 40; Staining and Labeling; Temperature

Topics: Agglutination; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cytochalasin B; Ferritins; Fibroblasts; Lectins; Ligands; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Microscopy, Electron; Staining and Labeling; Trypsin

Topics: Agglutination; Animals; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; DNA, Neoplasm; Fibroblasts; Mice; Microscopy, Electron; Ruthenium; Simian virus 40; Staining and Labeling; Surface Properties; Thymidine; Tritium

Topics: Agglutination Tests; Animals; Antigens, Viral; Cell Fusion; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Colchicine; Concanavalin A; Defective Viruses; Dose-Response Relationship, Drug; Floxuridine; Gammaretrovirus; Genetic Variation; Helper Viruses; Leukemia Virus, Murine; Mice; Moloney murine leukemia virus; Rats; RNA-Directed DNA Polymerase; Thymine Nucleotides; Tritium; Viral Interference; Virus Replication

Topics: Cell Transformation, Neoplastic; Concanavalin A; Hodgkin Disease; Humans; Immune Adherence Reaction; Immunity, Cellular; Lectins; Leukemia, Lymphoid; Lymphoma, Non-Hodgkin; Mitogens; T-Lymphocytes

Topics: Adenoviridae; Animals; Carbohydrates; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Embryo, Mammalian; Epithelial Cells; Fibroblasts; Formaldehyde; Histocytochemistry; Microscopy, Electron; Neoplasms, Experimental; Peroxidases; Receptors, Drug; Staining and Labeling

Topics: Animals; Binding Sites; Cell Aggregation; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Endoplasmic Reticulum; Female; Lectins; Lymphatic Metastasis; Male; Microscopy, Electron; Neoplasm Transplantation; Neoplasms, Experimental; Oncogenic Viruses

Topics: Agglutination; Animals; Ascitic Fluid; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Formaldehyde; Glutaral; Glycine max; Lectins; Lymphoma; Mice; Moloney murine leukemia virus; Plant Lectins; Polysaccharides; Protein Binding; Time Factors; Triticum; Tritium

The strength of attachment of normal and transformed baby hamster kidney cells was markedly increased when attached cells were treated with concanavalin A (Con A). The cells became less sensitive to detachment by physical shear or by treatment with trypsin or EDTA; however, their morphology, as observed by phase contrast microscopy, did not change. The effects of Con A were prevented by the simultaneous addition of either D-glucose or alpha-methyl-D-glucoside with the Con A. Also addition of these reagents to the attached cells after Con A treatment partially reversed the effects caused by Con A. Pre-treatment of the culture flasks with Con A before cell attachment resulted in an increase in the strength of cell attachment to the culture flasks as compared to untreated controls.

Topics: Animals; Cell Adhesion; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Edetic Acid; Glucose; Kidney; Methylglycosides; Stress, Mechanical; Time Factors; Trypsin

We have investigated the relationship of concanavalin. A binding to the cell surface of normal and transformed cells and the subsequent agglutination of the transformed cells. At room temperature almost no differences could be detected in agglutinin binding between transformed and untransformed cells. At 0 degrees C, however, where endocytosis was negligible, the transformed cells bound three times more agglutinin. However, transformed cells and trypsin-treated normal cells do not agglutinate at 0 degrees C although the amounts of agglutinin bound at 0 degrees C are sufficient to permit agglutination when such cells are shifted up to room temperature. Both transformed and trypsin-treated normal cells show a marked increase in agglutination at 15 degrees C as compared to agglutination at 0 degrees C. From this, as well as the observation that mild glutaraldehyde fixation of the cell surface inhibited agglutination but not agglutinin binding, it was concluded that concanavalin A-mediated cell agglutination requires free movement of the agglutinin receptor sites within the plane of the cell surface.

Topics: Animals; Antigen-Antibody Reactions; Binding Sites, Antibody; Cell Adhesion; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Endocytosis; Glutaral; Histological Techniques; Mice; Phosphatidylcholines; Temperature; Tritium; Trypsin

A COMPARISON IS MADE OF THE ULTRASTRUCTURE OF THE CELL PERIPHERY IN THREE CLONED CELL LINES: untransformed Balb/c 3T3 cells, SV40-transformed Balb/c 3T3 cells, and revertant cells obtained from the transformed cell line by a selection technique utilizing concanavalin A. Both thin-section and surface replication techniques are used for in situ examination of the cell lines. Microfilaments, 70 A in diameter (called alpha filaments), are abundant in untransformed and revertant cell lines, particularly in the anterior expansions of the cells, which tend to have many microvilli and small pseudopodia. Alpha filaments are diminished in the anterior expansions of transformed cells, which contain large blunt pseudopodia and relatively few microvilli. Surface replicas confirm the impression gained from thin sections that transformed cells have a greater proportion of their cell surface involved in bulging pseudopodia than either untransformed or revertant cells. Since alpha filaments are shown to bind heavy meromyosin and are similar to F-actin, these filaments are thought to be important in cell motility. These observations suggest that a close relationship exists between decreased alpha filaments, bulging pseudopodia, and loss of contact inhibition of movement in transformed cells.

Topics: Aneuploidy; Animals; Cell Line; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Contact Inhibition; Endoplasmic Reticulum; Fibroblasts; Glycerol; Histological Techniques; Mice; Mice, Inbred BALB C; Microscopy; Microscopy, Electron; Myofibrils; Pseudopodia; Ribosomes; Simian virus 40

A binding assay that shows consistent differences in the amounts of tritium-labeled concanavalin A that bind to normal and virally transformed cells was used to study the kinetic changes of cell surface in SV40-transformed 3T3 cells that express the transformed phenotype in a temperature-sensitive manner (tsSV3T3 cells). The increase in concanavalin A binding, which paralleled the appearance of the characteristics of transformed cells, was dependent on the synthesis of cellular DNA. In agreement with the results of binding studies, exponentially growing tsSV3T3 cells agglutinated at much lower lectin titers (concanavalin A as well as wheat-germ agglutinin) at 32 degrees than at 39 degrees .

Topics: Agglutination; Animals; Binding Sites; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; DNA; DNA, Neoplasm; Glucosamine; Haptens; Hydroxyurea; Kinetics; Lectins; Mice; Mutation; Phenotype; Simian virus 40; Temperature; Tritium

Topics: Animals; Carcinoma, Hepatocellular; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Clone Cells; Concanavalin A; Cycloheximide; Epithelial Cells; Epithelium; Fibroblasts; Lectins; Liver; Liver Neoplasms; Neoplasm Proteins; Peptide Hydrolases; Protein Biosynthesis; Proteins; Rats; Surface Properties

Topics: Agglutination; Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cholesterol; Concanavalin A; Contact Inhibition; Cyclic AMP; DNA; Fibroblasts; Humans; Hydrocortisone; L Cells; Male; Mice; Penis; Progesterone; Simian virus 40; Thymidine; Tritium

Topics: Agglutination Tests; Animals; Cell Line; Cell Transformation, Neoplastic; Cell-Free System; Cells, Cultured; Centrifugation, Density Gradient; Clone Cells; Concanavalin A; DNA Nucleotidyltransferases; DNA, Viral; Fibroblasts; Gammaretrovirus; Mice; Microscopy, Phase-Contrast; Nucleic Acid Hybridization; RNA, Viral; Time Factors; Transcription, Genetic; Tritium; Uridine; Virus Replication

Topics: Agglutination Tests; Animals; Carbon Isotopes; Carcinoma, Hepatocellular; Cell Count; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Fibroblasts; Hydrogen-Ion Concentration; Kinetics; Lectins; Liver; Liver Neoplasms; Magnesium; Mice; Mitosis; Peptide Hydrolases; Polyomavirus; Receptors, Drug; Simian virus 40; Temperature; Tritium

Topics: Agglutination; Animals; Antigens, Neoplasm; Butyrates; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cyclic AMP; Cytotoxicity Tests, Immunologic; Drug Synergism; Mice; Oncogenic Viruses; Rabbits; RNA Viruses; Theophylline

The topographical distributions of concanavalin A-binding sites on the surfaces of 3T3, proteasetreated 3T3, and simian virus 40-transformed 3T3 cultured mouse fibroblasts appear to be different, as shown by a shadow-cast replica technique using concanavalin A and a hemocyanin marker, or as shown previously on isolated membranes with concanavalin A coupled to ferritin. However, chemical fixation of cells before labeling with concanavalin A and hemocyanin, or labeling exclusively at 4 degrees , allows one to distinguish between inherent concanavalin A-binding-site topography and potential rearrangement of sites induced by the action of the multivalent concanavalin A molecule itself. The inherent distribution of binding sites on 3T3, protease-treated 3T3, and transformed cells is actually the same on all cells, i.e., dispersed and random. Treatment of unfixed transformed or protease-treated 3T3 cells, but not normal 3T3 cells, with concanavalin A and hemocyanin at 37 degrees (or at 4 degrees with subsequent warming to 37 degrees ), however, results in clustering of binding sites, presumably due to crosslinking of neighboring lectin-binding sites by the quadrivalent concanavalin A. Thus, the underlying difference between concanavalin A-binding sites on normal as compared with transformed or protease-treated normal cells lies not in the inherent topography of binding sites, but rather in the susceptibility of the sites to aggregation by concanavalin A. The latter may reflect an increased mobility of lectin-binding sites on transformed or protease-treated cells.

Topics: Animals; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Fibroblasts; Hemocyanins; Lectins; Mice; Protein Binding; Simian virus 40; Trypsin

Transformed fibroblasts had a low content of ATP when grown at a high cell density and a high content of ATP when grown at a low cell density. Concanavalin A agglutinated transformed cells with a low, but not those with a high, ATP content. Transformed cells with a high ATP content gained agglutinability after ATP depletion by inhibitors of the energy-generating systems, and those with a low ATP content lost their agglutinability after restoration of a high ATP content by glucose. Fixation of the surface membrane by formaldehyde, glutaraldehyde, or LaCl(3), inhibited agglutination of cells with an ATP content that allows agglutination. Normal fibroblasts grown at a high or a low cell density were not agglutinated by concanavalin A. Depletion of the cellular ATP content of normal cells induced agglutination only in cells grown at a high, but not at a low, cell density. A similar number of concanavalin A molecules was bound to the surface membrane of agglutinating and nonagglutinating fibroblasts. It is suggested that a high content of ATP inhibits the movement of concanavalin A binding sites, and that a low content of ATP allows, in transformed cells, a new distribution of binding sites to form the clusters required for cell agglutination. Agglutinability of transformed cells is determined by ATP content, and in normal cells changes in the content of ATP are by themselves not sufficient to induce agglutination. Transformed cells, therefore, do not have a control, presumably for membrane stability, that exists in normal cells.

Topics: Adenosine Triphosphate; Agglutination; Aldehydes; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Dinitrophenols; Fibroblasts; Fluorides; Formaldehyde; Glucose; Hydrazones; Lanthanum; Nitrites; Oligomycins; Polyomavirus; Simian virus 40

By treating populations of simian virus 40 (SV40)-transformed 3T3 cells with concanavalin A, variants have been isolated which are resistant to the killing action of the lectin. The variants (i) resemble 3T3 cells morphologically and in some of their growth characteristics; (ii) are not agglutinated by high concentrations of concanavalin A or wheat germ agglutinin, but can be rendered agglutinable by treatment with low concentrations of trypsin; (iii) bind the same number of concanavalin A molecules as 3T3 or SV3T3 cells; (iv) cannot be transformed by SV40 and are resistant to focus formation after infection with murine sarcoma virus; (v) contain SV40-specific T antigen and RNA and; (vi) yield wild-type SV40 virus after heterokaryon formation with BS-C-1 cells.

Topics: Agglutination Tests; Animals; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; DNA, Neoplasm; Drug Resistance; Fluorescent Antibody Technique; Genetic Variation; Haplorhini; Histocompatibility Antigens; Iodine Isotopes; Lectins; Mice; Microscopy, Phase-Contrast; Simian virus 40; Thymidine; Tritium; Viral Plaque Assay

Topics: Animals; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Embryo, Mammalian; Fibroblasts; Lectins; Mice; Mitosis; Protein Biosynthesis; Simian virus 40; Tritium

Topics: Agglutination; Animals; Cell Count; Cell Transformation, Neoplastic; Cells, Cultured; Chromosome Aberrations; Chromosomes; Clone Cells; Concanavalin A; Cricetinae; Dose-Response Relationship, Radiation; Embryo, Mammalian; Karyotyping; Leukemia, Radiation-Induced; Mitosis; Neoplasms, Radiation-Induced; Radiation Dosage; Radiation Effects; Radiation Genetics; Retroviridae

Topics: Agglutination Tests; Alpharetrovirus; Animals; Cell Line; Cell Transformation, Neoplastic; Chick Embryo; Clone Cells; Concanavalin A; Microscopy, Electron, Scanning; Microscopy, Phase-Contrast; Mutation; Rats; Temperature

Topics: Agglutination; Bucladesine; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; DNA; Herpesvirus 4, Human; Humans; Lymphocytes; Mitosis

Topics: Ammonium Sulfate; Animals; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Circular Dichroism; Concanavalin A; Cricetinae; Erythrocytes; Immune Sera; Lymphocytes; Microscopy, Electron; Phagocytosis; Rabbits; Serum Albumin, Bovine; Simian virus 40; Spleen

Topics: Animals; Biological Transport, Active; Carbon Radioisotopes; Cell Count; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chromatography, Paper; Clone Cells; Concanavalin A; Contact Inhibition; Genetic Variation; Glucose; Mice; Mice, Inbred BALB C; Simian virus 40; Tritium

Topics: Agar; Agglutination Tests; Animals; Ascitic Fluid; Cell Transformation, Neoplastic; Cells, Cultured; Chickens; Concanavalin A; DNA; Fluorescent Antibody Technique; Genetic Variation; Hot Temperature; Interferons; Macrophages; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Polyomavirus; Thymidine; Time Factors; Tritium

Topics: Adsorption; Agglutination; Binding Sites; Cell Fractionation; Cell Membrane; Cell Transformation, Neoplastic; Centrifugation, Density Gradient; Concanavalin A; Methods; Neoplasms, Experimental; Osteosarcoma; Peritoneal Neoplasms; Polyomavirus

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Concanavalin A; Cycloheximide; Cytarabine; Dactinomycin; Fibroblasts; Mutation; Puromycin; Temperature

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Diffusion; Energy Transfer; Fibroblasts; Fluorescence; Glutaral; Lymphocytes; Lymphoma; Protein Binding; Rats; Rotation; Simian virus 40; Sucrose; Trypsin

Topics: Animals; Avian Leukosis Virus; Binding Sites; Bone Marrow; Cell Aggregation; Cell Membrane; Cell Transformation, Neoplastic; Chickens; Concanavalin A; Erythrocytes; Ferritins; Glycoproteins; Microscopy, Electron; Nickel; Phenylhydrazines; Protein Binding; Radioisotopes; Receptors, Drug; Spectrophotometry; Tritium; Trypsin

Topics: Animals; Antigens; B-Lymphocytes; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Colchicine; Concanavalin A; Drug Resistance; Fluoresceins; Immunity, Cellular; Lectins; Microscopy, Electron; Microscopy, Fluorescence; Neoplasms; Ouabain; Phospholipids; Protein Binding; T-Lymphocytes

Topics: Agglutination; Animals; Carcinoma; Carcinoma, Squamous Cell; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fibroblasts; HeLa Cells; Humans; Kidney; Laryngeal Neoplasms; Lectins; Lung; Mice; Mouth Neoplasms; Neoplasms, Experimental; Trypsin

Topics: Agglutination Tests; Animals; Binding Sites; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fibroblasts; Goats; Lectins; Lymphocytes; Lymphoma; Mice; Models, Biological; Neoplasms, Experimental; Protein Binding; Rats

Topics: Adenoviridae; Agglutination Tests; Animals; Antigens, Neoplasm; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Cricetinae; Lectins; Neoplasm Transplantation; Trypsin; Tumor Virus Infections

Topics: Animals; Binding Sites; Cattle; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Clone Cells; Concanavalin A; Cricetinae; DNA, Viral; Fibroblasts; Hemagglutination; Humans; Microscopy, Electron; Oncogenic Viruses; Peroxidases; Rabbits; RNA, Viral; Temperature

Topics: Agglutination Tests; Animals; Carcinogens; Cell Division; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition; Cricetinae; Dextrans; Embryo, Mammalian; Female; Fibroblasts; Fibrosarcoma; Mice; Mitosis; Molecular Weight; Pregnancy; Sulfuric Acids; Time Factors

Topics: Animals; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Hemagglutination, Viral; Immune Sera; Lectins; Mammary Tumor Virus, Mouse; Methods; Mice; Mice, Inbred Strains; Microscopy, Electron; Phosphotungstic Acid; Protein Binding; Rauscher Virus; Staining and Labeling; Viral Proteins

Topics: Animals; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Embryo, Mammalian; Lectins; Microscopy, Electron; Peroxidases; Receptors, Drug; Staining and Labeling

Topics: Animals; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fibroblasts; In Vitro Techniques; Lectins; Microscopy, Electron; Polyomavirus; Rats; Receptors, Drug; Simian virus 40; Staining and Labeling

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Cell Line; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Erythrocytes; Humans; Lectins; Leukemia, Lymphoid; Liver; Lymphocytes; Mice; Neoplasms, Experimental; Neuraminidase; Plant Lectins; Rats; Spleen; Triticum; Trypsin

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Carbohydrates; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Fibroblasts; Lectins; Mutation; Phenotype; Surface Properties; Temperature

Transformed and nontransformed cells in tissue culture differ in their rate of uptake of certain nutrients, as determined by a polyester-coverslip technique. A 2.5- to 3.5-fold increased rate of uptake of alpha-aminoisobutyric acid, cycloleucine, and 2-deoxy-D-glucose was observed with polyoma virus-transformed baby hamster kidney (BHK) 21 cells and simian virus 40 (SV40)-transformed BALB/3T3 (mouse fibroblast) cells, compared to their nontransformed counterparts. Kinetic analysis suggested that the increased uptake by cells transformed with virus was associated with a 3-fold greater V(max), with no detectable changes in apparent K(m). Limited studies also revealed increased initial rates of uptake by murine sarcoma virus-transformed rat liver cells, as compared to the parental line. Exposure of cells to concanavalin A and wheat-germ agglutinin led to significant reductions in amino-acid uptake by both transformed and nontransformed cells; however, transformed cells showed a greater decrease in uptake after exposure to wheat-germ agglutinin. Increased initial rates of uptake of certain amino acids and sugars may be a feature common to transformed cells, compared to their parental control.

Topics: Amino Acids; Aminoisobutyric Acids; Animals; Arginine; Carbon Isotopes; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Hexoses; Kinetics; Lectins; Leucine; Mice; Neoplasms, Experimental; Polyomavirus; Rats; Simian virus 40

Topics: Agglutination Tests; Animals; Binding Sites; Cell Count; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Lectins; Polyomavirus; Trypsin

Topics: Agglutination; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Lectins; Neoplasm Transplantation; Neoplasms, Experimental; Nickel; Nitrosamines; Polyomavirus; Radioisotopes; Simian virus 40; Time Factors; Trypsin

Contact-inhibited variants have been isolated by treatment of simian virus 40 (SV40)-transformed Balb/c 3T3 cells (SVT2) with the plant lectin concanavalin A. These con A revertant cells exhibit the following properties: (i) they resemble 3T3 cells morphologically and grow to saturation densities which are similar to that of 3T3 cells; (ii) they synthesize the SV40-specific T antigen and yield infectious virus after fusion with permissive monkey cells; (iii) they contain a high sialic acid content similar to that of 3T3 cells and not to that of SVT2 cells; sialic acid composition was found to be independent of serum concentration; (iv) they contain more chromosomes with the average number in the tetraploid range than the SVT2 cells from which they were derived; and (v) SVT2 and revertant cells, confluent or subconfluent, produce more collagen than Balb/3T3 cells. The relationship of surface membrane properties to contact inhibition of growth and the mechanisms for generating revertant cells are discussed.

Topics: Agglutination; Animals; Antigens, Viral; Cell Fusion; Cell Line; Cell Transformation, Neoplastic; Chromosomes; Clone Cells; Collagen; Concanavalin A; Contact Inhibition; Floxuridine; Genetic Variation; Haplorhini; Lectins; Mice; Neuraminic Acids; Polyploidy; Proline; Simian virus 40; Tritium

Differentiated epithelial cells in contact-inhibited monolayers derived from adult rat liver have been transformed in vitro into epithelioid neoplastic cells under conditions of nutritional stress. The transformed cells maintain their differentiated quality and manufacture serum proteins. They differ from control cultures in the following properties: They are aneuploid, can be agglutinated by wheat-germ agglutinin and concanavalin A, can grow in suspension, and are able to form colonies in semisoft agar. There is no intercellular communication at permeable junctions between the cells; this is demonstrable by electrical measurement or by injection of fluorescein. The cells show invasiveness in culture, and are not inhibited by contact with normal cells. The characteristics of the hepatocytes after transformation in vitro resemble those of epithelioid cells derived from a transplantable hepatoma.

Topics: Agglutination; Aneuploidy; Animals; Blood Proteins; Carcinoma, Hepatocellular; Cell Adhesion; Cell Differentiation; Cell Line; Cell Transformation, Neoplastic; Clone Cells; Concanavalin A; Contact Inhibition; Culture Media; Epithelial Cells; Epithelium; Immunoelectrophoresis; Karyotyping; Lectins; Liver; Liver Neoplasms; Nutritional Physiological Phenomena; Rats; Stress, Physiological

Topics: Acetates; Animals; Calcium; Cell Adhesion; Cell Membrane Permeability; Cell Transformation, Neoplastic; Chelating Agents; Clone Cells; Concanavalin A; Contact Inhibition; Cytoplasm; DNA; DNA, Neoplasm; Edetic Acid; Glycols; Glycoproteins; Macromolecular Substances; Magnesium; Mice; Mice, Inbred Strains; Neoplasm Proteins; Proteins; RNA; RNA, Neoplasm; RNA, Transfer; Simian virus 40

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Carbohydrates; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Fibroblasts; Glycoproteins; Hyaluronic Acid; Hyaluronoglucosaminidase; Lectins; Molecular Weight; Mutation; Polyomavirus; Temperature

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Cell Line; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Hexosamines; Lectins; Mannose; Plant Lectins; Triticum; Trypsin

Topics: Avian Sarcoma Viruses; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Embryo, Mammalian; Fibroblasts; Histocytochemistry; Humans; Immunity, Cellular; Lectins; Microscopy, Electron; Peroxidases

Hybrids between mouse cells and simian virus 40 (SV40)-transformed rat cells were made, and their properties and chromosome constitution were investigated over many generations. Their hybrid nature was confirmed by enzyme studies. During a period of 1 year a loss of 10 to 20% of the total number of chromosomes was observed. The SV40 tumor antigen was present and remained present in the hybrids. The parental and hybrid cells were studied for agglutination with concanavalin A, for growth in soft agar, and for serum requirement. These growth and surface characteristics of the transformed cells appeared in the hybrids.

Topics: Agglutination; Animals; Antigens, Viral; Cell Line; Cell Transformation, Neoplastic; Chromosomes; Clone Cells; Concanavalin A; Culture Media; Electrophoresis; Fluorescent Antibody Technique; Hybrid Cells; Immune Sera; Indophenol; L-Lactate Dehydrogenase; Mice; Oxidoreductases; Phosphogluconate Dehydrogenase; Rats; Rats, Inbred Strains; Simian virus 40

Topics: Agglutination; Animals; Carcinoma; Carcinoma, Squamous Cell; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Cricetinae; Cycloheximide; Cytarabine; Floxuridine; Humans; Immune Sera; Kidney; Laryngeal Neoplasms; Lectins; Methods; Newcastle disease virus; Simplexvirus; Time Factors; Trypsin; Vitamin A

The binding of fluorescein-conjugated Concanavalin A to the cell surface has been studied in normal and transformed cells in interphase and mitosis. Binding to the cell surface was in the form of an incomplete ring of fluorescence, and the binding was inhibited by alpha-methyl-D-mannopyranoside. All the cells were fluorescent when treated with 25-250 mug/ml of fluorescent Concanavalin A. With 10 mug, the cells were all fluorescent after 30 min of binding, but after 0.5-5 min with 10 mug or 30 min with 1 or 2.5 mug, transformed interphase cells showed a higher percentage of cells with surface fluorescence than did normal interphase cells. Trypsinized normal and transformed interphase cells showed the same fluorescence. Binding with 2.5 mug at 4 degrees instead of at 24 degrees , gave a higher percentage of fluorescent cells with trypsinized than with untrypsinized transformed cells. Mitotic normal cells were similar to transformed interphase cells, whereas mitotic transformed cells were intermediate between normal and transformed interphase cells. The results indicate that the use of low concentrations of fluorescent Concanavalin A can show differences in surface fluorescence between normal and transformed, interphase and mitotic, and trypsinized and untrypsinized cells. It is suggested that the observed differences in fluorescence can be explained by differences in affinity of the lectin binding sites and/or differences in the clustering of sites.

Topics: Animals; Binding Sites; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Embryo, Mammalian; Fluoresceins; Fluorescence; Glycosides; Kinetics; Lectins; Mice; Microscopy, Fluorescence; Mitosis; Neoplasms, Experimental; Nitrosamines; Polyomavirus; Protein Binding; Simian virus 40

Topics: Agglutination; Animals; Carbohydrate Metabolism; Carbohydrates; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Fibroblasts; Lectins; Mice; Simian virus 40; Trypsin

Topics: Agglutination; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Glycosides; Hemagglutination; Hemagglutination Inhibition Tests; Herpesvirus 4, Human; Humans; Iodine Isotopes; Lectins; Lymphocytes; Mannose; Receptors, Drug

Topics: Agglutination Tests; Animals; Binding Sites, Antibody; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Concanavalin A; Cricetinae; Culture Techniques; Cycloheximide; Glucosamine; Glycosides; Kidney; Lectins; Newcastle disease virus; Plant Lectins; Polyomavirus; Radiation Effects; Triticum; Ultraviolet Rays; Viral Proteins; Virus Replication

The sensitivity of transformed 3T3 fibroblasts to agglutination by concanavalin A is reduced by alkaloids that bind specifically to protein subunits of microtubules.

Topics: Agglutination; Animals; Cell Line; Cell Transformation, Neoplastic; Colchicine; Concanavalin A; Erythrocytes; Hemadsorption; Hemagglutination; Lectins; Mice; Simian virus 40; Vinblastine

Topics: Agglutination; Animals; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Gammaretrovirus; Mice; Simian virus 40; Temperature

Topics: Agglutination; Animals; Cats; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Centrifugation, Density Gradient; Clone Cells; Concanavalin A; Cricetinae; Embryo, Mammalian; Gammaretrovirus; Humans; Leukemia Virus, Murine; Mice; Moloney murine leukemia virus; Rats; Rhabdomyosarcoma; RNA Viruses; Thymus Gland; Tritium; Uridine; Viral Plaque Assay; Virus Replication

Topics: Agglutination; Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Hemadsorption; Kidney; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Orthomyxoviridae; Simian virus 40

Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Adenylyl Cyclases; Animals; Binding Sites; Cell Count; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition; Culture Media; Cyclic AMP; Fibroblasts; Mice; Potassium; Simian virus 40; Time Factors; Tritium; Trypsin

Topics: Adenoviridae Infections; Agglutination; Animals; Ataxia Telangiectasia; Carcinoma; Cell Transformation, Neoplastic; Chromosome Aberrations; Chromosome Disorders; Chromosome Mapping; Chromosomes; Colonic Neoplasms; Concanavalin A; Culture Techniques; Humans; Immunoglobulin A; Immunoglobulin E; Nucleic Acid Hybridization; Rats

Topics: Agglutination; Animals; Binding Sites; Cell Line; Cell Membrane; Cell Survival; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Drug Resistance; Genetic Variation; Lectins; Leucine; Ornithine; Peptides; Simian virus 40

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Cell Line; Cell Transformation, Neoplastic; Chickens; Concanavalin A; Fibroblasts; Humans; Lectins; Neoplasms, Experimental; Plant Lectins; Plants, Edible; Rats; Sarcoma, Avian

Topics: Agglutination; Animals; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Concanavalin A; Cricetinae; Kidney; Lectins; Newcastle disease virus; Trypsin

Topics: Animals; Binding Sites; Cell Adhesion; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Culture Media; Culture Techniques; Lectins; Methods; Microscopy, Electron; Microscopy, Phase-Contrast

Topics: Agglutination; Animals; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Chromosome Aberrations; Concanavalin A; Cricetinae; Embryo, Mammalian; Karyotyping; Mitosis; Neoplasms, Experimental; Polyploidy; Sex Chromosomes; Surface Properties; Trisomy

A cell line, derived from a spontaneous equine connective tissue tumor (equine sarcoid), has been established. The morphological and growth characteristics indicative of malignant transformation of the cells include a disoriented, rapid growth and loss of contact inhibition. Further evidence of transformation is the agglutination of these cells by concanavalin A and their ability to divide in semisolid media.

Topics: Agar; Agglutination Tests; Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Culture Media; Culture Techniques; Horses; Male; Skin Neoplasms; Trypsin

Topics: Agglutination Tests; Animals; Carcinogens; Cell Aggregation; Cell Transformation, Neoplastic; Cells, Cultured; Chromosomes; Concanavalin A; Culture Techniques; Cyclic N-Oxides; Diploidy; Liver; Mitosis; Neoplasm Transplantation; Neoplasms, Experimental; Nitro Compounds; Photomicrography; Quinolines; Rats; Rats, Inbred Strains; Transplantation, Homologous

Topics: Agglutination Tests; Animals; Cell Fusion; Cell Line; Cell Transformation, Neoplastic; Chromosome Aberrations; Clone Cells; Concanavalin A; Cricetinae; Hybrid Cells; Karyotyping; Neoplasm Transplantation; Transplantation, Heterologous

Topics: Agglutination Tests; Animals; Carbohydrates; Cell Aggregation; Cell Line; Cell Membrane; Cell Survival; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Depression, Chemical; Female; In Vitro Techniques; Lectins; Rats; Sarcoma, Yoshida; Structure-Activity Relationship

Topics: Animals; Binding Sites; Cell Aggregation; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Lectins; Lymphoma; Mice; Neoplasms, Experimental; Protein Binding

Topics: Agglutination; Animals; Binding Sites; Cell Aggregation; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Chromatography, Affinity; Chromatography, DEAE-Cellulose; Concanavalin A; Cricetinae; Fucose; Glycoproteins; Mice; Plant Lectins; Plant Proteins; Protein Binding; Rats; Seeds; Structure-Activity Relationship; Trypsin

Topics: 3T3 Cells; Animals; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Ferritins; Mice; Mice, Inbred BALB C; Neoplasms; Plant Lectins; Protein Binding

Topics: Adenoviridae; Agglutination; Animals; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Culture Techniques; Fibroblasts; Lectins; Liver; Nickel; Oncogenic Viruses; Polyomavirus; Protein Binding; Radioisotopes; Rats; Tritium

Topics: Agglutination; Animals; Binding Sites; Cell Line; Cell Transformation, Neoplastic; Chromatography, DEAE-Cellulose; Chromatography, Gel; Concanavalin A; Cricetinae; Culture Techniques; Fibroblasts; Iodine Isotopes; Lectins; Oncogenic Viruses; Plant Lectins; Polyomavirus; Protein Binding; Temperature; Triticum; Trypsin

The carbohydrate-binding protein, Concanavalin A (Con A), binds to glucose- or mannose-like sites on the cell-surface membrane. Unless the cells are treated with trypsin, this protein agglutinates malignantly transformed cells, but not normal cells. The transformed cells were agglutinated at 24 degrees C but not at 4 degrees C. Transformed and normal cells treated with trypsin were agglutinated at both 24 degrees C and 4 degrees C with high concentrations of Con A (500 mug/ml), but only at 24 degrees C with low concentrations (5 mug/ml). The same number of Con A molecules were bound to normal and transformed cells at both temperatures. The results indicate that the site for Con A on the surface membrane contains two activities, a component that binds Con A molecules (B) and a component that determines agglutination (A). B is not temperature sensitive and is active in normal and transformed cells, whereas A, which is temperature sensitive, is in an active form only in transformed cells. A can be activated by trypsin, and the increased activity per cell allows agglutination at 4 degrees C with a high, but not with a low, concentration of Con A. Agglutination of transformed cells by wheat-germ agglutinin, which binds to N-acetyl-D-glucosamine-like sites, and by soybean agglutinin, which binds to N-acetyl-D-galactosamine-like sites, was not temperature sensitive. Thus, the temperature-sensitive component A is specific for Con A, and malignant transformation of normal cells, which results in agglutinability by Con A, is associated with the activation of a specific temperature-sensitive activity on the surface membrane.

Topics: Agglutination Tests; Animals; Avian Sarcoma Viruses; Binding Sites; Cattle; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Haplorhini; Hexoses; Lectins; Lymphoma; Moloney murine leukemia virus; Neoplasms, Experimental; Nitrosamines; Polyomavirus; Protein Binding; Temperature

We have measured the quantitative binding of the radioactively labeled agglutinin (125)I-concanavalin A to normal mammalian cells and simian virus 40- and polyoma virus-transformed cells from tissue culture. Parallel measurements of the amount of (125)I-concanavalin A necessary to cause agglutination of the cells in suspension were carried out. The transformed and nontransformed cells used for these experiments show large differences in their ability to be agglutinated by (125)I-concanavalin A. However, these cell lines have the same number of specific binding sites and similar affinities for the agglutinin whether transformed, trypsinized, or nontransformed. We conclude that the differential capacity of concanavalin A to agglutinate transformed cells relative to normal cells does not result from differences in the number of binding sites between the two types of cells.

Topics: Agglutination; Animals; Binding Sites; Calcium; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Culture Media; Fibroblasts; Iodine Isotopes; Kidney; Lectins; Magnesium; Mice; Plant Lectins; Plants, Edible; Polyomavirus; Simian virus 40; Trypsin

Topics: Agglutination; Animals; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Kidney; Lectins; Mice; Polyomavirus; Virus Cultivation

Topics: Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Lectins; Mice; Neoplasm Transplantation; Neoplasms, Experimental

Topics: Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition; Neoplasms; Neoplasms, Experimental; Surface Properties

A surface receptor for an agglutinin, exposed in transformed but not in normal cells, arises in normal mouse cells during lytic infection by polyoma virus. The structural change in the surface membrane characteristic of transformed cells and of cells productively infected by wild type virus fails to occur in normal mouse cells infected by mutants of the virus that are unable to cause transformation. The exposure of the receptor site by wild type virus is reversibly blocked by inhibitors of DNA synthesis.

Topics: Agglutination Tests; Animals; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Contact Inhibition; DNA; Haptens; Kidney; Lectins; Mice; Mutation; Polyomavirus; Puromycin; Radiation Effects; Ultraviolet Rays