concanavalin-a and Neoplasm-Metastasis

The liver remains a hematopoietic organ after birth and can produce all leukocyte lineages from resident hematopoietic stem cells. Hepatocytes produce acute phase proteins and complement in bacterial infections. Liver Kupffer cells are activated by various bacterial stimuli, including bacterial lipopolysaccharide (LPS) and bacterial superantigens, and produce interleukin (IL)-12. IL-12 and other monokines (IL- 18 etc.) produced by Kupffer cells activate liver natural killer (NK) cells and NK1.1 Ag+ T cells to produce interferon-gamma and thereby acquire cytotoxicity against tumors and microbe-infected cells. These liver leukocytes and the T helper 1 immune responses induced by them thus play a crucial role in the first line of defense against bacterial infections and hematogenous tumor metastases. However, if this defense system is inadequately activated, shock associated with multiple organ failure takes place. Activated liver NK1.1 Ag+ T cells and NK cells also cause hepatocyte injury. NK1.1 Ag+ T cells and another T-cell subset with an intermediate T-cell receptor, CD 122+CD8+ T cells, can develop independently of thymic epithelial cells. Liver NK cells and NK1.1 Ag+ T cells physiologically develop in situ from their precursors, presumably due to bacterial antigens brought from the intestine via the portal vein. NK cells activated by bacterial superantigens or LPS are also probably involved in the vascular endothelial injury in Kawasaki disease.

Topics: Animals; Antigens, Bacterial; Cell Lineage; Child, Preschool; Concanavalin A; Gram-Positive Bacteria; Humans; Intestinal Absorption; Killer Cells, Natural; Kupffer Cells; Lipopolysaccharides; Liver; Liver Circulation; Lymphocyte Activation; Lymphokines; Macrophage Activation; Mice; Mice, SCID; Mucocutaneous Lymph Node Syndrome; Multiple Organ Failure; Neoplasm Metastasis; Neoplastic Cells, Circulating; Peritonitis; Receptors, Antigen, T-Cell; Receptors, Antigen, T-Cell, gamma-delta; Shock, Septic; Shwartzman Phenomenon; Superantigens; T-Lymphocyte Subsets; T-Lymphocytes, Cytotoxic; Th1 Cells

An artificial receptor for proMMP-9 was created by fusing tissue inhibitor of MMP-1 (TIMP-1) with type II transmembrane mosaic serine protease (MSP-T1). Expression of MSP-T1 in 293T cells induced binding of proMMP-9, which was processed by MMP-2 activated by membrane type 1 MMP (MT1-MMP). HT1080 cells transfected with the MSP-T1 gene produced activated MMP-9 in collagen gel, and addition of proMMP-2 to the culture augmented it, which resulted in intensive collagen digestion. These cells metastasized into chick embryonic liver more than control cells. Treatment of HT1080 cells with concanavalin A in the presence of exogenous proMMP-2 induced activation of not only proMMP-2 but also proMMP-9. Knockdown of MT1-MMP or TIMP-2 expression with siRNA suppressed activation of both proMMP-2 and proMMP-9. Transfection of TIMP-1 siRNA suppressed cell binding and activation of proMMP-9, but not proMMP-2 activation. Knockdown of a disintegrin and metalloproteinase 10 (ADAM10) expression reduced cell binding and processing of proMMP-9. These results suggest that proMMP-9, which binds to a receptor complex containing TIMP-1 and ADAM10, is activated by the MT1-MMP/MMP-2 axis, and MMP-9 thus activated stimulates cellular proteolysis and metastasis.

Topics: ADAM10 Protein; Amyloid Precursor Protein Secretases; Animals; Cell Line; Chick Embryo; Concanavalin A; Enzyme Activation; HEK293 Cells; Humans; Matrix Metalloproteinase 1; Matrix Metalloproteinase 14; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Membrane Proteins; Neoplasm Metastasis; Neoplasms; Protein Binding; Recombinant Fusion Proteins; RNA Interference; RNA, Small Interfering; Serine Endopeptidases; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinase-2

The production of cytokines by peripheral blood cells and biopsy specimens of tumors stimulated by polyclonal activators (PAs) was evaluated in 34 patients with invasive ductal breast carcinoma using enzyme-linked immunosorbent assay (ELISA). Positive correlation between the stimulation index of polyclonal activators (SIPA) for IL-18 production by the tumor and the relative content of poorly differentiated cells was revealed. The latter, in turn, was positively correlated with the numbers of normal and pathologic mitoses and the degree of malignancy. Cancer cells can produce IL-18, which is involved in the process of angiogenesis, stimulates invasion and metastasis. Decrease in SIPA for the production of IL-6 and GCSF by peripheral blood cells could serve as an indicator of malignant progression in invasive ductal breast carcinoma.

Topics: Blood Cells; Carcinogenesis; Carcinoma, Ductal, Breast; Concanavalin A; Enzyme-Linked Immunosorbent Assay; Female; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-18; Interleukin-6; Lipopolysaccharides; Mitosis; Neoplasm Invasiveness; Neoplasm Metastasis; Neovascularization, Pathologic; Phytohemagglutinins

Upregulated expression of eN has been found in the highly invasive human melanoma cell lines but neither in melanocytes nor in primary tumor cells. Membrane proteins associated with cell adhesion and metastasis: alpha5-, beta1-, beta3-integrins, and CD44 were elevated gradually in accordance with increasing metastatic potential. alphav-integrin was seen mostly in aggressive melanomas. The expression of eN correlated with a number of metastasis-related markers and thus may have a function in the process. eN activity went parallel with its amount in all cells. Concanavalin A strongly inhibited the enzyme in a noncompetitive way. Clustering of eN protein in overexpressing cells by ConA-treatment increased the enzyme association with the heavy cytoskeletal complexes. A similar shift towards cytoskeletal fractions took also place with other membrane proteins coexpressed with eN. This ConA-induced association may reflect a putative interaction of eN with physiological ligand, that upon interaction, aggregates protein components of lipid rafts and triggers signaling pathway that may be intrinsically involved in cell-stroma adhesion.

Topics: 5'-Nucleotidase; Antigens, Neoplasm; Cell Adhesion; Cell Line, Tumor; Cluster Analysis; Concanavalin A; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Hyaluronan Receptors; Integrin alphaV; Melanoma; Membrane Microdomains; Neoplasm Metastasis

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

We attempted to investigate if the in vivo administration of concanavalin A (Con A), a potent T cell stimulator, would render anti-metastatic activity in hosts. Assays of activity were performed 20 days after iv inoculation of two clones of the B16 melanoma, B16-H (H-2+, highly metastatic), B16-L (H-2-, low metastatic), or 3LL cells into C57BL mice by enumerating lung colonies. In some experiments, hosts treated with anti-asialo GM1 Ab were used to evaluate effector mechanisms other than NK cells. While the injection of Con A alone had no significant effect on anti-metastatic activity, in nonimmunized hosts the effect by Con A was displayed when the mice were preimmunized with B16-H cells but not in those immunized with B16-L cells. Immunization with B16-H or B16-L cells alone resulted in the generation of killer cells with promiscuous lytic activity and induced an anti-metastatic effect against B16-H, B16-L, and 3LL cells. Con A treatment significantly augmented the killer activity of spleen cells of mice preimmunized with B16-H cells but not of those immunized with B16-L cells. The effectors from mice immunized with B16-H alone or given both Con A and B16-H were mainly of Thy 1+ Lyt2+ asialo GM1- cells, on the other hand, those from mice immunized with B16-L cells expressed asialo GM1 antigen. We showed the efficacy of Con A on the anti-metastatic effect in relation to the host immune response.

Topics: Animals; Concanavalin A; Female; H-2 Antigens; Immunization, Passive; Interleukin-2; Killer Cells, Natural; Lymphocyte Activation; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Neoplasm Metastasis; Phenotype; T-Lymphocytes

The effect of surgery on peripheral blood mononuclear cell responsiveness to mitogens and suppressor cell (SC) activity assessed in a concanavalin A (ConA) assay were studied in patients with stage 0 and stage III-IV cancer. Patients were exposed to a similar surgical trauma the same type of anaesthesia, and to no pre- and early postoperative radio- or chemotherapy. A more pronounced postoperative decrease in the lymphocyte count, responsiveness to phytohemagglutinin (PHA) and ConA, and in the SC activity was found in the nonadvanced than advanced cancer group. These findings point to an impaired mobilization and distribution capacity of circulating lymphocytes in patients with advanced neoplastic disease.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Carcinoma in Situ; Cells, Cultured; Colonic Neoplasms; Concanavalin A; Female; Humans; Leukocyte Count; Leukocytes, Mononuclear; Lymphatic Metastasis; Lymphocytes; Middle Aged; Neoplasm Metastasis; Phytohemagglutinins; Stomach Neoplasms; T-Lymphocytes, Regulatory; Uterine Neoplasms

This study has examined cells from naturally-occurring murine mammary tumours to ascertain whether cell surface glycoproteins play a significant role in colonisation of the lungs after intravenous inoculation. It was found that gel electrophoretic analysis of membrane extracts and lectin adsorption studies did not reveal any consistent differences in glycoprotein composition of cells from tumours which can heavily colonise the lungs relative to ones from tumours which cannot do so or to cells from pulmonary metastases. Also, alteration of structural and functional properties of surface glycoproteins by treatment with succinylated lectins or with drugs such as tunicamycin and swainsonine, which inhibit glycosylation of membrane proteins, had no specific effects on metastatic colonisation of the lungs. Tunicamycin apparently decreased capability to form experimental metastases but also diminished tumourigenicity on subcutaneous inoculation, although it did not affect tumour cell viability in vitro. This information supports earlier studies from this laboratory involving enzymic digestion of the surface of living tumour cells before inoculation and demonstrates that the pulmonary colonisation capability of these mammary tumour cells can withstand global disorganisation of membrane glycoprotein structure and composition. This implies that either the surface glycoproteins are not important in the colonisation process, or that these tumour cells have great capability for rapid repair of their surfaces. It is concluded that a clear answer to whether surface glycoprotein composition has a decisive role in pulmonary colonisation by these mammary tumour cells requires introduction of stable heritable traits into tumour cell populations by genetic manipulation.

Topics: Alkaloids; Animals; Cell Membrane; Concanavalin A; Glycoproteins; Lung Neoplasms; Mammary Neoplasms, Experimental; Membrane Proteins; Mice; Mice, Inbred C3H; Neoplasm Metastasis; Neoplasm Proteins; Swainsonine; Tunicamycin; Wheat Germ Agglutinins

It has repeatedly been proposed that fibrin plays a role in tumor growth and metastasis. Among tumor cell products or activities which may promote clot formation, cancer procoagulant (CP), a direct activator of coagulation factor X, has been suggested to be selectively associated with the malignant phenotype. We report here the enzymatic and immunological identification of this cysteine proteinase procoagulant in extracts and cells from human melanoma. CP activity was independent of both the intrinsic and extrinsic pathways of blood coagulation, using factor IX and factor VII deficient plasmas, and was inhibited by the cysteine proteinase inhibitors iodoacetamide and HgCl2. CP activity was detectable in extracts and cell suspensions from all 32 patients studied and was higher in extracts from metastases (14.8 +/- 3.9 units/mg protein) than from the primary tumors (3.7 +/- 1.0 units/mg protein). CP activity was not affected by an anti-apoprotein III antibody or by concanavalin A, a known inhibitor of thromboplastin. In contrast, no CP activity or antigen was detected in extracts from six benign melanocytic lesions. The procoagulant activity was dependent on factor VII and was inhibited by anti-apoprotein III antibody and by concanavalin A, properties that suggest that the procoagulant was tissue thromboplastin. These data indicate that CP can be expressed by human tumor cells and that, among melanotic lesions, its presence is associated with the malignant phenotype and its activity is particularly high in metastatic cells.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Blood Coagulation Factors; Concanavalin A; Cysteine Endopeptidases; Endopeptidases; Female; Humans; Male; Melanoma; Mercuric Chloride; Middle Aged; Neoplasm Metastasis; Neoplasm Proteins

Quantitative studies on the binding of concanavalin A (Con A) and wheat-germ agglutinin (WGA) to a series of rat hepatocarcinoma metastatic variants revealed a positive correlation between the amount of cell-surface-bound lectin and lung colonization potential. Scatchard analysis of Con A and WGA binding to 10 individual clones isolated from a subcutaneous (s.c.) tumor transplant and to tumor-cell isolates from 10 individual spontaneous lung metastases from the same animal showed diverse binding characteristics for these cell populations. Nevertheless, the expression of Con A receptor sites accurately predicted the lung colonization potential of 3 isolates from the lung metastases. Higher lectin binding curves were observed for the clones from the subcutaneous tumor than for the isolates from lung metastases. These data suggest that a high Con-A binding potential is indicative of a high lung colonization potential for these hepatocarcinoma cells, but that this phenotype may be rapidly lost during tumor outgrowth in the lungs. The binding of tumor cells to vascular endothelial cell monolayers was inhibited in the presence of Con A; however, no inhibition was observed with 2 other lectins. Attachment of tumor cells to endothelial cell monolayers was also inhibited by the monosaccharides methyl alpha-D-mannopyranoside and N-acetyl-D-galactosamine. Other monosaccharides tested did not alter the attachment of tumor cells to endothelial cell monolayers.

Topics: Animals; Blood Vessels; Carbohydrates; Cell Adhesion; Cell Line; Concanavalin A; Endothelium; Glycopeptides; Iodine Radioisotopes; Lectins; Lung Neoplasms; Mannose; Monosaccharides; Neoplasm Metastasis; Neoplasms, Experimental; Rats; Skin Neoplasms; Wheat Germ Agglutinins

We have studied some cellular characteristics and the transplantability of a newly induced squamous cell carcinoma, Sq1-SC, in comparison with the ascites-transformed subline of the same tumor, Sq1-AA. We could demonstrate that the AA tumor differed from the SC tumor in the pattern of intravenously induced 'experimental metastases'. The SC tumor preferentially gave rise to extrapulmonary tumor colonies ('metastases'), while the AA tumor exclusively gave rise to lung colonies. Comparison with the ascites tumor growing in solid form subcutaneously (AS tumor) shows that the enzymatic treatment, which is necessary to bring solid tumors into viable and dissociated suspensions, can have a decisive influence on tumor cell lodgement in vessels and metastasis.

Topics: Animals; Ascites; Blood Coagulation; Carcinoma, Squamous Cell; Cell Separation; Concanavalin A; Electrophoresis; Female; Lectins; Methylcholanthrene; Mice; Mice, Inbred CBA; Neoplasm Metastasis; Neoplasm Transplantation; Trypsin; Wheat Germ Agglutinins

The capacity of inbred W/Fu rats bearing syngeneic colon carcinomas to generate interleukin(s) (IL) was studied during primary tumor growth, after tumor resection, and during postresection immunotherapy. During local tumor growth, there was a significant decrease in the capacity of the host's adherent mononuclear cells to generate IL-1 and of peripheral blood mononuclear cells to generate IL-2 (16.6 and 23%, respectively, when compared to control animals; P less than .01). The presence of regional metastases or large primary tumor burden resulted in a further sharp fall in IL generation (0.9 and 10% for IL-1 and IL-2, respectively, when compared to control animals; P less than .01). With the use of three different doses of tumor inoculum, inhibition of IL generation was shown to occur when tumors were barely palpable. Decrease in IL correlated with tumor growth and not with the initial number of tumor cells injected. Tumor resection resulted in a rise in IL-2 generation from 36 to 64% of control animals' levels. Postresection immunotherapy with the use of an active specific immunization protocol successfully modulated IL-2 production to normal in animals protected from tumor recurrence. Animals that developed recurrent tumors despite immunization exhibited a continued inability to generate IL (mean values of IL-2 production compared to controls: 184% in animals free of recurrence after immunotherapy, 1% in animals developing recurrent tumors after immunotherapy; P less than .01). These results suggested that alterations in IL generation may lead to immune unresponsiveness during tumor growth. Active specific immunotherapy protecting animals from recurrence after primary tumor resection may be predicated on the successful modulation of IL level generation by host immunocytes.

Topics: Adenocarcinoma; Animals; Cell Division; Cells, Cultured; Colonic Neoplasms; Concanavalin A; Immunotherapy; Interleukin-1; Interleukin-2; Male; Monocytes; Neoplasm Metastasis; Neoplasm Recurrence, Local; Neoplasm Transplantation; Rats; Rats, Inbred Strains

MDW4, a wheat germ agglutinin resistant mutant of the murine tumor line MDAY-D2, expresses abnormal asparagine-linked oligosaccharides, is less metastatic when injected intravenously, and is hypersensitive to natural killer (NK) lysis in vitro. To determine whether these phenotypes may be related, variants of the YAC-1 lymphoma and a YAC-1 X MDAY-D2 hybrid line were compared for sensitivity to four different lectins and to NK cell lysis in vitro. A relationship between sensitivity to concanavalin A (Con A) and NK cell lysis in vitro was observed. Although no single plasma membrane glycoprotein separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with 125I-labeled Con A correlated with NK and Con A sensitivities of the cell lines, a relationship between these phenotypes and the collective 125I-Con A staining intensity on the gels was apparent. In a more direct test of carbohydrate recognition by NK cells, specific glycopeptide structures isolated from tumor cells and added to the NK cell assay in microM quantities were found to inhibit tumor cell lysis. Thus, a subset of asparagine-linked oligosaccharides, including high mannose and some incomplete complex structures on a number of cell surface glycoproteins, appears to be recognized as part of the target structures for NK cell lysis. The administration of polyinosinic:polycytidylic acid stimulated splenic NK activity in vivo but had no effect on the growth of the NK-resistant MDAY-D2 cells. However, the low tumorigenicity of MDW4 cells injected intravenously was reduced further by pretreating the mice with polyinosinic:polycytidylic acid, which indicated a role for NK cells in the elimination of circulating tumor cells expressing high mannose and/or incomplete complex asparagine-linked oligosaccharides.

Topics: Animals; Binding, Competitive; Cell Line; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Glycopeptides; Glycoproteins; Killer Cells, Natural; Lectins; Membrane Proteins; Mice; Neoplasm Metastasis; Neoplasm Proteins; Neoplasms, Experimental; Receptors, Concanavalin A; Structure-Activity Relationship

The concanavalin A binding characteristics of serum alpha-fetoprotein (AFP) were investigated in patients with primary hepatocellular carcinoma and hepatic secondaries using affinity column chromatography and radioimmunoassay. The primary hepatocellular carcinoma (n = 21) was associated with a median concanavalin A non-reactive AFP fraction of 7.4% (range 1.6 - 18.8) while the hepatic secondaries (n = 8) had a median concanavalin A non-reactive AFP fraction of 50.7% (range 26.6 - 91.7). A simple diagnostic test for differentiating between the two groups of patients is proposed.

Topics: Adolescent; Adult; Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Chromatography, Affinity; Concanavalin A; Diagnosis, Differential; Female; Humans; Liver Neoplasms; Male; Middle Aged; Neoplasm Metastasis

Cellular glycoprotein carbohydrate chains of B16 melanoma sublines of various metastatic colonization capacities were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and direct lectin staining, combined with chemical modification of carbohydrate chains in situ. For these studies, we utilized B16 sublines selected for low (B16-F1) or high lung (B16-F10), high brain (B16-B15b), or high ovary (B16-O13) colonization properties, or high tissue invasiveness in vitro (B16-BL6). The major B16 cell surface sialoglycoproteins were of Mr approximately 115,000, approximately 90,000, approximately 82,000, and 60,000 to 65,000, and were detectable by periodate NaB3H4 labeling and binding of 125I-wheat germ agglutinin (WGA). Terminal sialic acid residues in the carbohydrate chains were responsible for WGA binding, since chemical removal of sialic acid prevented WGA labeling of the glycoproteins. However, removal of sialic acid residues followed by Smith degradation resulted in reappearance of WGA-binding sites on these sialoglycoproteins, indicating that the carbohydrate chains possessed at least one branching point at an outer alpha-mannosyl residue. This structural feature was further indicated by the failure of 125I-Lens culinaris hemagglutinin to bind to these sialoglycoproteins. The fact that the carbohydrate residues of the Mr approximately 115,000, approximately 90,000, and approximately 82,000 sialoglycoproteins were of the complex type was confirmed by their reactivity with 125I-Ricinus communis agglutinin I, which preferentially binds to Gal leads to GlcNAc sequences after removal of sialic acid in situ. In contrast, 125I-peanut (Arachis hypogaea) agglutinin, specific for Gal leads to GalNAc sequences, failed to bind to the major WGA-reactive sialoglycoproteins, but strongly interacted after removal of sialic acid with Mr approximately 51,000 and approximately 56,000 glycoproteins from sublines B16-F1, -F10, and -BL6 and with a Mr approximately 63,000 glycoprotein from sublines B16-F10, -BL6, -O13, and -B15b. Thus, the small, mucin-type carbohydrate chains were expressed almost exclusively on these lower Mr sialoglycoproteins, and very little on the Mr approximately 82,000, approximately 90,000, and approximately 115,000 sialoglycoproteins. Differences in lectin binding to glycoproteins were observed with different sublines. These glycoproteins included: (a) a WGA-binding Mr 60,000 to 75,000 sialoglycoprotein prominent

Topics: Animals; Carbohydrates; Cell Line; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Glycoproteins; Lectins; Melanoma; Mice; Neoplasm Metastasis; Peanut Agglutinin; Plant Lectins

Reduced cellular immune response is well documented in patients with advanced breast cancer. To investigate immunocompetence at the time of diagnosis, 104 patients with breast cancer staged according to the TNM classification were studied preoperatively and compared with 95 age matched healthy women. Tests of blood mononuclear leukocytes included lymphocyte and monocyte counts, determination of rosette forming T (SER +) and B (MER +) lymphocytes, T lymphocyte subsets defined with monoclonal antibodies (Leu-1, Leu-2a, Leu-3a) and with lectin fractionation (soybean agglutinin, SBA), lymphocyte transformation tests with PHA and ConA and colony formation of T cells in agar (TL-CFC). Two age groups (A: 30-50, B: 51-70 years) and the different tumor stages (I-IV) were analyzed. Patients and controls did not differ in absolute numbers of lymphocytes, T and B cells. In patients of group B the absolute number of monocytes was slightly increased in stages II and III and significantly in stage IV (p less than 0.025). Similarly, the lymphocyte response to PHA was significantly reduced in stage IV group B only (p less than 0.05). ConA induced lymphocyte proliferation and TL-CFC capacity were not different in patients and controls. In the small number of patients and age matched controls, in whom T lymphocyte subsets were determined, the relative numbers of T cells with helper or suppressor phenotype as defined with Leu-3a, Leu-2a, or SBA were similar. In conclusion, in breast cancer, at the time of diagnosis, blood T lymphocyte populations and functions are not altered except in elderly patients with disseminated disease. The monocytosis and reduced PHA responsiveness observed in the latter group may be related phenomena.

Topics: Adult; Aged; Breast Neoplasms; Concanavalin A; Female; Humans; Immunity, Cellular; Leukocyte Count; Lymphocyte Activation; Lymphocytes; Middle Aged; Monocytes; Neoplasm Metastasis; Neoplasm Staging; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Tumor pairs, selected on the basis of their different capacities to metastasize in vivo (SP73/AS and ASML from the rat, Eb/ESb from the mouse), have been assayed for their membrane associated actin through the DNase inhibition assay. It is found that, provided inhibitions per cell are corrected for the influence of gross heterogeneities in size distributions, the more metastatic tumor cells have significantly higher DNase I inhibitions than their less invasive counterparts. This observation, which extends our previous study of normal recirculating lymphocytes, is rationalized by postulating a participation of these actin pools to a property critical for both normal recirculation and metastatic spreading, arguments are presented which favor cell surface deformability as a possible candidate.

Topics: Actins; Adenocarcinoma; Animals; Concanavalin A; Deoxyribonuclease I; Endodeoxyribonucleases; Female; Fibrosarcoma; Lymphoma; Male; Neoplasm Metastasis; Neoplasms; Rats

When the tumor-bearing leg of C57BL/6J mice was amputated 16 days after SC inoculation of 10(6)B16 melanoma cells, all the amputated mice died of pulmonary metastases. Transfer of lungs from the amputated to normal syngeneic mice revealed tumor cells in the lungs just after amputation. Repeated weekly injections of BCG and irradiated tumor cells, beginning 24 h after amputation of the tumor-bearing limb, prolonged the survival only of mice presensitized to BCG. Injections of BCG or irradiated melanoma cells alone, or neuraminidase- and mitomycin C-treated tumor cells or of Levamisole had no effect, but injections of ConA-coated tumor cells slightly prolonged the survival of the amputated mice. Both BCG and B16 cells induced humoral and cell-mediated immunity but there was no cross-reactivity between BCG and B16 cells.

Topics: Animals; Antibody Formation; BCG Vaccine; Concanavalin A; Disease Models, Animal; Immunity, Cellular; Immunization; Immunotherapy; Levamisole; Lung Neoplasms; Melanoma; Mice; Neoplasm Metastasis; Neoplasms, Experimental; Neuraminidase

Rat 13762NF mammary adenocarcinoma cell surface glycoproteins from s.c. tumor- or lung metastases-derived cell clones of differing spontaneous metastatic potentials were examined for their relationship to metastasis. After treatment with neuraminidase, lectin-binding assays showed that highly metastatic clone MTLn3 cells express approximately twice the quantity of peanut agglutinin (PNA) binding sites (approximately 2.3 X 10(8) sites/cell) than clones of lower metastatic potential. However, the number of wheat germ agglutinin (WGA)-binding sites on the various cell clones decreased slightly as the metastatic potential of the clones increased. The quantities of concanavalin A (conA)-binding sites were similar (approximately 1.7 X 10(8) sites/cell) in all cell clones and growth conditions. Glycoprotein analysis was performed by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate (SDS-PAGE) and subsequent staining with 125I-labeled lectins. SDS-PAGE gels stained with 125I-labeled conA revealed mainly one glycoprotein (Mr approximately 150 kD), and the amounts of this glycoprotein did not correlate with metastasis. Differences in WGA-binding glycoproteins were detected between s.c. tumor- and lung metastases-derived cell clones. Several desialylated glycoproteins were detected with 125I-labeled PNA after SDS-PAGE, and the labeling intensity of one (Mr approximately 580 kD) correlated with the metastatic potentials of the various cell clones. This high Mr galactoprotein was further analyzed by [3H]glucosamine metabolic labeling, solubilization, sequential gel filtration, and chondroitinase ABC treatment prior to SDS-PAGE. The 580 kD galactoprotein was expressed in increased amounts on the more highly metastatic clones. Chemical labeling of cell surface sialic acid residues using periodate treatment followed by [3H]borohydride reduction showed an additional change in a major sialoglycoprotein (Mr approximately 80 kD), which decreased in labeling intensity on clones of increasing metastatic potential. The results suggest quantitative changes in cell surface glycoproteins rather than major qualitative alterations are associated with differences in the metastatic behavior of 13762NF tumor cell clones.

Topics: Adenocarcinoma; Animals; Arachis; Binding Sites; Clone Cells; Concanavalin A; Glycoproteins; Lectins; Mammary Neoplasms, Experimental; Membrane Proteins; Mice; Neoplasm Metastasis; Neoplasm Proteins; Peanut Agglutinin; Plant Lectins; Sialoglycoproteins; Surface Properties; Wheat Germ Agglutinins

Cell suspensions from the R 3230 AC rat mammary adenocarcinoma, when injected intravenously into F344 rats, invariably produce multiple lung foci within 10 days. We compared the colonization potential of cultures obtained from these foci and from cell populations exposed to 100 micrograms/ml medium of both concanavalin A and wheat germ agglutinin for 5 passages with the original cell line. Plasminogen activator activity (PAA) was determined in all three cell subpopulations, using S2251 (KABI) as chromogenic substrate. All cell lines retained their ability to grow after subcutaneous implant. The lectin resistant variant was found to have lost its capacity to nidate in the lung completely and also had the lowest PAA. In contrast, the cell population derived from the lung foci ranked highest in PAA.

Topics: Adenocarcinoma; Animals; Cell Line; Concanavalin A; Female; Lectins; Mammary Neoplasms, Experimental; Neoplasm Metastasis; Neoplasm Transplantation; Plasminogen Activators; Rats; Rats, Inbred F344; Wheat Germ Agglutinins

In an attempt to magnify differences in the immune responses of potentially immunosuppressed cancer patients and normal controls, an assessment was made on the effects of the competitive inhibitor alpha-methyl-D-mannoside on the concanavalin A (Con A)-induced blastogenic responses of lymphocytes from each of these populations. Lymphocytes from breast cancer patients with metastatic disease were significantly deficient in their capability to undergo blast transformation regardless of whether the monosaccharide inhibitor was added to the assay cultures. In contrast, lymphocytes from breast cancer patients who did not display metastatic disease were capable of normal blastogenic responses to Con A. The addition of alpha-methyl-D-mannoside to lymphocyte cultures caused a significantly greater inhibition of the blastogenic responses of these patients' cells as compared to cells of normal controls. Thus the monosaccharide seems to serve as a useful reagent for optimizing differences between lymphocyte blastogenic responses of normal donors and those of immunodepressed donors. The results suggest that lymphocytes from breast cancer patients without clinically evident metastases possess some modification of their cell membrane. One possibility discussed was that the number or distribution of receptors for Con A on the membrane of lymphocytes of these patients is deficient.

Topics: Binding, Competitive; Breast Neoplasms; Cell Membrane; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Drug; Female; Humans; Lymphocyte Activation; Lymphocytes; Methylglycosides; Methylmannosides; Middle Aged; Neoplasm Metastasis; Receptors, Concanavalin A

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

A neutral protease has been extracted from the media of cultured metastatic tumor cells and purified approximately 1000 times after sequential ammonium sulfate fractionization, concanavalin A column chromatography, and molecular sieve chromatography. The protease has an apparent molecular weight of 70 000--80 000, is inactive at acid pH, requires trypsin activation, and is inhibited by ethylene-diaminetetraacetic acid but not by phenylmethanesulfonyl fluoride, N-ethylmaleimide, or soybean trypsin inhibitor. The enzyme produces specific cleavage products for both chains of pro type IV collagen isolated without pepsinization and apparently cleaves at one point in a major pepsin-extracted chain of placenta type IV collagen. The partially purified enzyme fails to significantly degrade other collagens or fibronectin under digestion conditions in which specific reaction products are produced for type IV collagen. The existence of this enzyme is significant since previously described animal collagenases fail to degrade type IV collagen. Such a type IV specific collagenase could play a role in tumor invasion and may be secreted by other cells such as endothelial cells, epithelial cells, and immune cells.

Topics: Animals; Concanavalin A; Mice; Microbial Collagenase; Molecular Weight; Neoplasm Metastasis; Sarcoma, Experimental; Substrate Specificity

We have studied several metastatic variant cell lines derived from a common clonal origin and their transformed and untransformed parental cell lines. A number of in vitro characteristics were examined for each tumor line and these properties were correlated with the ability of the tumor cells to form pulmonary nodules in an experimental metastasis assay. Direct correlations with metastatic behavior in the lung colony assay were found to exist with the amount of cell-bound Concanavalin A and the procoagulant activities of cell lysates. In vitro parameters that did not correlate with the metastatic phenotype were: population doubling times in culture, saturation density achieved in culture, the number of colony-forming cells shed from confluent cultures, rates of cellular attachment to homotypic or heterotypic cell monolayers, plasminogen-activator production and procoagulant activity produced in serum-free conditioned medium.

Topics: Animals; Cell Line; Clone Cells; Concanavalin A; Neoplasm Metastasis; Phenotype; Plasminogen Activators; Tumor Cells, Cultured

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

Lymphocyte blast transformation of 23 melanoma patients was compared to that of 22 healthy persons after stimulation with the mitogens PHA, ConA, and PWM. Transformation rate of lymphocytes in microcultures was measured following 3H-Thymidin uptake in a Liquid-Scintillation-Counter. Calculations were based on analyses of variance. There was no significant difference in blastogenetic response between the 2 groups (patients and controls), but there were differences between the used mitogens.

Topics: Adolescent; Adult; Aged; Concanavalin A; Female; Humans; Lymphocyte Activation; Lymphocytes; Male; Melanoma; Middle Aged; Neoplasm Metastasis; Phytohemagglutinins; Pokeweed Mitogens; Skin Neoplasms

The fraction of carcinoembryonic antigen preparations not bound by concanavalin A was studied. A significant portion of this nonbound fraction of low antigenic activity was shown to be mucopolysaccharides by gas chromatography-mass spectrometry, cellulose acetate strip electrophoresis, and depolymerization by bovine testicular hyaluronidase.

Topics: Carcinoembryonic Antigen; Chemical Phenomena; Chemistry; Chromatography, Gas; Colonic Neoplasms; Concanavalin A; Glucuronates; Glycosaminoglycans; Humans; Liver Neoplasms; Neoplasm Metastasis

An alpha-feto-protein (AFP) is present in many mammals, in birds, and in sharks during development. The AFP present in different species have similar physicochemical properties and often have common antigenic determinants. Their study, both in health and disease, has provided a useful model for the understanding of other phase-specific antigens and the activation of the genes which control their synthesis. In the human fetus, the level of AFP falls with increasing maturity. The more sensitive methods of detection have disclosed that this fetal protein persists in trace amounts throughout life and its level increases in maternal blood during pregnancy. The principal sites of synthesis are the fetal liver and in some mammals, the yolk sac splanchnopleur. In humans as well as in mice and cows, it is notable that the synthesis of AFP is increased in liver cancer cells and that high levels of this protein are present in serum. Elevated values of AFP have also been detected in human subjects with undifferentiated tumours of the testis and ovary. A fall to normal levels has been noted in cases of complete remission after surgery and a return to high levels in patients who develop metastases. In some patients with hepatitis a temporary rise in the level of AFP has also been observed. In recent years, the detection of high levels of AFP in amniotic fluid has proved to be of great value for the prenatal diagnosis of neural-tube defects. Abnormal levels have also been found in the amniotic fluid or in maternal serum in cases of spontaneous abortion. Such measurements are now being assessed as a methodof monitoring abnormal pregnancy.

Topics: alpha-Fetoproteins; Amniotic Fluid; Anencephaly; Animals; Antigen-Antibody Reactions; Carcinoma, Hepatocellular; Concanavalin A; Cystic Fibrosis; Down Syndrome; Female; Fetal Proteins; Gastrointestinal Neoplasms; Gestational Age; Hepatitis; Humans; Immunologic Techniques; Infant, Newborn; Liver; Liver Neoplasms; Metabolism, Inborn Errors; Neoplasm Metastasis; Neoplasms, Experimental; Pregnancy; Spinal Dysraphism; Teratoma

The carcinoembryonic antigen (CEA) active glycoproteins from perchloric acid extract of liver-metastasized primary colon tumor have been separated by concanavalin A Sepharose (Con A Sepharose) chromatography. The CEA activities separated by Con A Sepharose chromatography were designated as loosely bound and tightly bound which, respectively, eluted on the Con A Sepharose column between 0.12 and 0.15 M and 0.3 M alpha-methylmannose in a linear gradient of alpha-methylmannose. Further purification of these activities by Sephadex G-200, Bio-Gels A-1.5m and P-300 yielded two variants of glycoproteins (B1 and C2) with CEA activity. Both purified preparations of CEA had similar immunochemical properties. Their A280/A260 ratios were 1.30 and 1.56, respectively. The purified loosely bound CEA (B1) had immunological, chromatographic, and electrophoretic properties similar to those of 125I-CEA, whereas the tightly bound CEA (C2) had a lower molecular weight (120,000 to 140,000). Further, specificity to these two CEA's was established by their reactions in immunoelectrophoresis with preparations of specific goat anti-CEA anti-serum obtained from other investigators. The results indicate the practical use of Con A Sepharose affinity chromatography for the separation and characterization of glycoprotein tumor antigens.

Topics: Carcinoembryonic Antigen; Chromatography, Affinity; Chromatography, Gel; Colonic Neoplasms; Concanavalin A; Epitopes; Glycoproteins; Humans; Neoplasm Metastasis; Radioimmunoassay

Topics: Animals; B-Lymphocytes; Binding Sites, Antibody; Bone Marrow Cells; Cell Differentiation; Concanavalin A; Immunity, Cellular; Lung Neoplasms; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred C57BL; Mononuclear Phagocyte System; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Radiation Chimera; Rats; Rats, Inbred Lew; Spleen; T-Lymphocytes; Thymectomy; Thymidine; Transplantation, Heterologous; Tritium

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Carcinoma, Hepatocellular; Chromatography, Affinity; Colonic Neoplasms; Concanavalin A; Freeze Drying; Glutaral; Humans; Liver Neoplasms; Neoplasm Metastasis

Carcinoembryonic antigen (CEA) was purified from tumor tissue under mild conditions at neutral pH by a procedure that utilized affinity chromatography on concanvalin A. Further purification by gel filtration provided CEA in 10 to 20% yield and 10% purity. Antibody to this preparation was rendered specific for CEA by adsorption on a column of normal liver proteins bound to Sepharose. On reaction by immunodiffusion against a crude tumor extract, the adsorbed antibody produced two precipitin lines, of which one was relatively weak. These two precipitin lines fused completely with the two respective lines produced by antibody to perchloric acid-treated CEA. The major antigen found in crude tumor extracts and in CEA preparations purified at neutral pH was nearly undetectable in perchloric acid extracts of tumor homogenates. Further investigations showed that 60 to 70% of the CEA in crude tumor extracts and in preparations isolated at netural pH is destroyed and/or becomes insoluble acidic conditions.

Topics: Carcinoembryonic Antigen; Chromatography, Affinity; Chromatography, Gel; Colonic Neoplasms; Concanavalin A; Humans; Hydrogen-Ion Concentration; Immune Sera; Immunodiffusion; Liver Neoplasms; Neoplasm Metastasis; Neoplasms

The use of Con A-Sepharose affinity chromatography for preparation of CEA from two metastatic liver tumors resulted in a separation of two species of CEA. One is concanavalin A-reactive CEA (CEA-M): the other is Con A-nonreactive CEA (CEA-P). Both CEA-M and CEA-P were glycoproteins and have identical antigenicity. However, 4 samples of CEA-M and CEA-P subfractions differed in their protein:carbohydrate ratios. The yield of CEA-M was greater than that of CEA-P. In electrophoresis, both CEA-M and CEA-P migrated at the region of beta-globulin of human blood serum. The isoelectric points of 4 samples of CEA-M and CEA-P subfractions from two different tumor sources differed from each other. In these preparations CEA-M usually showed a larger value of isoelectric point than CEA-P. Ultracentrifugal analysis of these four preparations revealed only a single peak, except CEA-P in case 1. Antigenic activity of CEA-M was almost completely destroyed by beta-N-acetylhexosaminidase treatment but only partly digestion with pronase. A possibility was suggested that N-acetylglucosamine at nonreducing terminal(s) is essential for the antigenic determinant groups of CEA molecule.

Topics: Binding Sites; Carbohydrates; Carcinoembryonic Antigen; Chromatography, Affinity; Concanavalin A; Hexosamines; Humans; Immunodiffusion; Immunoelectrophoresis; Liver Neoplasms; Molecular Weight; Neoplasm Metastasis; Pancreatic Neoplasms; Protein Binding; Rectal Neoplasms

Monosaccharide compositions of Con A-reactive CEA (CEA-M) and Con A-nonreactive CEA (CEA-P) separated from two different samples of CEA were analysed by gas liquid chromatography. It was revealed that all CEA subfractions possessed N-acetylglucosamine, fucose, and galactose residues. One out of 4 subfractions did not contain sialic acid and another one lacked glucose in its carbohydrate moiety. N-acetylgalactosamine was not detected in measurable amount in any of the 4 subfractions. A large amount of mannose was found in CEA-M, but only a small amount in CEA-P.

Topics: Acetylgalactosamine; Acetylglucosamine; Binding Sites; Carcinoembryonic Antigen; Chromatography, Affinity; Concanavalin A; Fucose; Galactose; Glucosamine; Humans; Liver Neoplasms; Mannose; Neoplasm Metastasis; Pancreatic Neoplasms; Protein Binding; Rectal Neoplasms; Sialic Acids

The carcinoembryonic antigen (CEA) of normal and pathological tissue extracts was separated into two variants, Concanavalin A reactive (CEAr) and non-reactive CEA (CEAn) by affinity chromatography on Con A Sepharose columns. CEAr was the quantitatively predominant variant. CEAn varied in concentration between 0.2 and 6 percent of the total CEA activity. The affinity of CEAn for anti-CEA antibodies was significantly lower than that of CEAr. Pooled extracts of primary adenocarcinomas of the colon contained CEAn in the lowest concentration and with the least affinity for antibodies. It is suggested that a deficiency and/or steric blocking of alpha-D-mannopyranosyl residues in CEAn reduce the affinities for both antibodies and Con A.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Chromatography, Affinity; Colonic Neoplasms; Concanavalin A; Fetus; Genetic Variation; Humans; Liver; Liver Cirrhosis; Liver Neoplasms; Lung; Neoplasm Metastasis; Organ Specificity; Radioimmunoassay; Spleen

A strain-specific transplantable melanoma (S-91) growing progressively in DBA/1 mice and metastasizing selectively to the lungs was maintained for 16 days in organ culture before being grafted to syngeneic (DBA/1) and allogeneic (BALB/c and C57BL/6) recipients. The cultured S-91 grew progressively in the syngeneic mice and to a moderate degree in the allogeneic strains; it showed an increased tendency to metastasize in both the DBA/1 and C57BL/6 recipients. Heterophilic cytoagglutination assays of cultured S-91 were less apt to aggregate in the presence of concanavalin A than were their noncultured counterparts, which suggested alteration of the plasma membrane. Organ culture explantation appeared to alter phenotypically the cell-surface membrane and thus increase the cell's ability to metastasize while possibly reducing the immunogenicity of the cultured tumor cells.

Topics: Agglutination Tests; Animals; Cells, Cultured; Concanavalin A; Lung Neoplasms; Melanoma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred DBA; Neoplasm Metastasis; Neoplasms, Experimental

Topics: Animals; Body Weight; Carcinoma; Concanavalin A; Female; Genotype; Graft vs Host Reaction; Hybridization, Genetic; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mononuclear Phagocyte System; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Organ Size; Spleen; T-Lymphocytes; Time Factors

Topics: Carcinoembryonic Antigen; Colonic Neoplasms; Concanavalin A; Electrophoresis; Fluorescent Antibody Technique; Humans; Immunodiffusion; Liver Neoplasms; Mannose; Neoplasm Metastasis

Topics: Agglutination Tests; Animals; Cell Line; Ceramides; Clone Cells; Concanavalin A; Cricetinae; Embryo, Mammalian; Fibroblasts; Glycolipids; Injections, Subcutaneous; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Phosphatidylcholines; Phosphatidylethanolamines; Phosphatidylinositols; Sphingomyelins

Topics: Concanavalin A; Congresses as Topic; Corynebacterium; Humans; Neoplasm Metastasis; Neuraminic Acids; Neuraminidase; Piperazines; Surface-Active Agents; Vaccination