concanavalin-a and Ovarian-Neoplasms

Cell immortalization technique has been an important method to establish cell lines with expected phenotype. However, there was few report about the establishment of immortalized human ovarian carcinoma cell lines. The purpose of this study was to establish an immortalized human ovarian sarcomatoid carcinoma cell line, and to investigate its biological characteristics.. A specimen derived from metastatic tumor of a human ovarian sarcomatoid carcinoma in abdominal wall was obtained and cultured in vitro. The 10th passage was transfected with SV40 T gene, colonies were isolated by G418 selection and expanded to immortalized cell lines. The morphology of the cells was observed by microscope and electromicroscope, Growth curve, karyotype analysis, culturing in soft agar, nude mice transplantation, immunohistochemistry, etc. were used to investigate its biologic characteristics. The biologic characteristics of BUPH: OVSC-2 were compared with its original cells.. After transfection and selection, an immortalized ovarian carcinoma cell line BUPH: OVSC-2 was established. The cell line has been maintained for over 90 passages. The H&E stain finding of transplanted tumor showed that the cells were morphologically sarcomatoid. However, the transmission electron microscopic observation exposed its epithelial origin. The cells grew exuberantly and had high malignant characteristics. Comparative studies revealed that the biological properties of BUPH: OVSC-2 and its original cells were identical.. BUPH: OVSC-2 was demonstrated as an immortalized human ovarian sarcomatoid carcinoma cell line with high malignancy. It retains similar properties of their parental cells and could be a useful model for the study of human ovarian sarcomatoid carcinoma.

Topics: Cell Division; Cell Size; Chromosomes; Concanavalin A; Female; Humans; Ovarian Neoplasms; Transfection; Tumor Cells, Cultured

Cancer patients generally exhibit circulating tumor-reactive immunoglobulins; however, these antibodies fail to eradicate tumors or prevent their progression. This study identifies and characterizes an aberrant tumor-reactive IgG population present in women with ovarian cancer.. In this pilot study, IgG was isolated from the sera of women with advanced-stage ovarian cancer (stages III and IV, n = 62) and age-matched female volunteers (n = 50) by affinity chromatography. These IgGs were characterized on the basis on their aberrant binding to concanavalin A affinity columns. Subsequently, the concanavalin A-binding moiety was localized following IgG fragmentation, analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and characterized by oligosaccharide profiling.. The level of concanavalin A-binding IgG in our control population was 8.9 +/- 2.9%, whereas in ovarian cancer patients, the level of concanavalin A-binding IgG was 38.8 +/- 7.4%. In the patients with ovarian cancer, 87.5 +/- 5.7% of the tumor-reactive IgG was demonstrated to be concanavalin A-binding. Based on oligosaccharide profiling of the fragmented concanavalin A-binding IgG, the aberrant lectin binding appeared to be the consequence of altered glycosylation of one of the two Fc chains.. While our previous studies have identified the presence of circulating IgG reactive with specific tumor-associated antigens and its association with poor prognosis, this report demonstrated the presence of an aberrantly glycosylated IgG population in cancer patients. This altered IgG appeared to be the primary class of tumor-reactive antibodies in these women.

Topics: Antibodies, Neoplasm; Antigens, Neoplasm; Chromatography, Affinity; Concanavalin A; Female; Humans; Immunoglobulin G; Middle Aged; Neoplastic Cells, Circulating; Oligosaccharides; Ovarian Neoplasms; Pilot Projects; Receptors, Concanavalin A

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

Metastatic dissemination of epithelial ovarian carcinoma occurs primarily through exfoliation of cells from the primary tumor, with subsequent implantation, invasion, and growth throughout the organs within the peritoneal cavity. Previous studies have suggested a role for matrix metalloproteinases (MMPs), particularly MMP-2, in ovarian cancer invasion and metastasis. To characterize further the role of MMPs and their inhibitors in ovarian carcinoma, in this study the production and activation of MMPs by short-term primary cultures of human ovarian epithelial carcinoma cells were analyzed. We report that MMP-2 is the predominant gelatinolytic MMP secreted by primary ovarian cancer cells derived from both ovarian tumors and ascites fluid. Furthermore, zymographic analysis demonstrated that MMP-2 is present in conditioned media in both the latent and activated forms, indicating that primary ovarian cancer cells catalyze proMMP-2 activation. Presence of a proMMP-2 activator was confirmed by immunohistochemistry and immunoprecipitation studies which found membrane-type 1 MMP (MT1-MMP) in the membranes of unstimulated cells and levels of both MT1-MMP and tissue inhibitor of metalloproteinases-2 (TIMP-2) were enhanced by culturing cells in the presence of concanavalin A. In addition, interaction of MMP-2 with the ovarian carcinoma cell surface resulted in a 2.5- to 5-fold increase in invasiveness. These data suggest that MT1-MMP-catalyzed activation of proMMP-2 may play a physiologic role in intraperitoneal invasion of ovarian carcinoma cells.

Topics: Carcinoma; Collagen; Concanavalin A; Culture Media, Conditioned; Drug Combinations; Female; Gelatinases; Humans; Immunohistochemistry; Laminin; Matrix Metalloproteinase 2; Matrix Metalloproteinases, Membrane-Associated; Metalloendopeptidases; Neoplasm Invasiveness; Ovarian Neoplasms; Proteoglycans; Tumor Cells, Cultured

Proteolytic enzymes could be very important in spread of cancer, but the role of the body's natural inhibitors of these enzymes in this process is unknown. One such inhibitor is the serum glycoprotein, alpha-1-proteinase inhibitor (API). In previous studies we showed that the fucose-specific lectin, lotus tetragonolobus, extracted high amounts of API in cancer when patients were unresponsive to treatment. The aim of this study was to determine whether the carbohydrate structure of API is altered in cancer. API was isolated from the sera of healthy women and women with breast or ovarian cancer. By means of high-performance anion-exchange chromatography, cancer API was shown to contain more fucose and less N-acetylglucosamine than healthy API. Further investigation of the purified specimens using a lectin-binding assay suggested that the cancer API was less branched and contained more alpha 2-6 and less alpha 2-3 sialic acid. Observations from both methods were consistent with an increase in bi-antennary chains terminating in alpha 2-6 sialic acid and possibly more alpha 1-6 fucose in the core of the unit. These distinctive changes could have important consequences for the function of API in cancer and may help to develop more precise markers for monitoring pathological progression in this disease.

Topics: Adenocarcinoma, Mucinous; Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin; Breast Neoplasms; Carbohydrate Conformation; Carbohydrates; Concanavalin A; Female; Fucose; Glucans; Humans; Lectins; Middle Aged; Ovarian Neoplasms; Phytohemagglutinins; Plant Lectins; Ribosome Inactivating Proteins; Serine Proteinase Inhibitors; Sialic Acids

The concentration of haptoglobin in sera of healthy women and patients with non-malignant and malignant ovarian tumours was measured by two methods, i.e. complex formation with haemoglobin and complex formation with concanavalin A. High correlation (r = 0.74) between both the methods was found in the group of healthy women, but correlation coefficients were much lower in the group of non-malignant and malignant tumours (r = 0.42 and 0.37, respectively). Direct determinations of haptoglobin, and calculation of the ratio of haptoglobin bound to concanavalin A to haptoglobin bound to haemoglobin revealed statistically significant differences among the examined groups. Comparison of two methods of haptoglobin quantitation suggest that processes connected with ovarian disorders may alter the glycosylation of haptoglobin oligosaccharide chains.

Topics: Adult; Biomarkers, Tumor; Concanavalin A; Female; Haptoglobins; Humans; Middle Aged; Ovarian Diseases; Ovarian Neoplasms; Reference Values

The lectin histochemistry of ovarian mucinous cystadenoma was studied using a panel of lectins comprising Triticum vulgaris, Lotus tetragonolobus, Arachis hypogaea, Griffonia simplicifolia I, concanavalin A, and Dolichus biflorus. All 23 cases examined in this study stained extensively with T. vulgaris. Of all the lectins, D. biflorus was the least reactive. Concanavalin A stained mainly the perinuclear zone, whereas the other lectins frequently stained the cytoplasm, glycocalyx, and extracellular luminal mucin. This pattern of lectin reactivity resembles endocervical more than intestinal epithelium and suggests that ovarian mucinous cystadenomas are of müllerian nature. The lack of difference in lectin reactivity between cystadenomas with and without goblet/Paneth cells supports a common histogenetic origin of both groups.

Topics: Concanavalin A; Cystadenoma; Female; Histocytochemistry; Humans; Lectins; Mucins; Ovarian Neoplasms; Peanut Agglutinin; Plant Lectins; Staining and Labeling; Wheat Germ Agglutinins

To establish whether alpha-fetoprotein (AFP) produced in the early post-treatment phase of a patient with a germ cell tumour of the testis or the ovary originates from the tumour or is due to an underlying disturbance in liver function, the binding of AFP to concanavalin A (Con A) was investigated as a discriminative variable. A two-step assay is described that can distinguish the type of AFP produced at levels as low as 10 ng/ml. A Con A-binding ratio of 12-43% was found in the patients with disseminated germ cell tumours and in patients with AFP-positive gastrointestinal carcinomas. AFP from the liver gives ratios below 10%.

Topics: alpha-Fetoproteins; Biomarkers, Tumor; Carcinoma, Hepatocellular; Concanavalin A; Female; Gastrointestinal Neoplasms; Humans; Liver Neoplasms; Male; Methods; Neoplasms, Germ Cell and Embryonal; Ovarian Neoplasms; Protein Binding; Testicular Neoplasms

The sensitivity of freshly derived human ovarian tumors (FOT) to various allogeneic cytotoxic effector cells stimulated by recombinant interleukin 2 (rIL-2), recombinant interferon alpha 2 (rIFN-alpha 2), OK-432, and concanavalin A was examined using the 51Cr release assay. Peripheral blood lymphocytes (PBL) of normal female donors were used as source of effector cells. Incubation of PBL with these biological response modifiers for 24 h generated effector cells with high natural killer activity, and only 20% (1/5) of the FOT examined were susceptible to lysis. By contrast, 83% (5/6) of the FOT were sensitive to lymphokine-activated killer (LAK) cells generated by rIL-2. OK-432 and concanavalin A activation of PBL also generated cytotoxic cells, though the cytotoxic activity against FOT was much less than that obtained by LAK cells. The addition of OK-432 to LAK culture medium containing rIL-2 generated effector cells with higher cytotoxicity against FOT than cultures with IL-2 alone. However, the addition of rIFN-alpha 2 in LAK culture medium resulted in the generation of effector cells with lower cytotoxicity. The addition of rIL-2, rIFN-alpha 2, or OK-432 to LAK cells during the in vitro cytotoxicity assay had no significant effect. When FOT target cells were pretreated with OK-432 they became more sensitive to LAK than nontreated tumor cells. However, pretreatment with rIL-2 or rIFN-alpha 2 did not influence cytolysis. These results suggest that the generation of LAK cells in vitro using rIL-2 plus OK-432 may be a more effective way to prepare these cells for adoptive immunotherapy in the treatment of ovarian cancer.

Topics: Concanavalin A; Cytotoxicity, Immunologic; Drug Combinations; Female; Humans; Immunologic Factors; Interferon Type I; Interleukin-2; Killer Cells, Lymphokine-Activated; Killer Cells, Natural; Lymphocyte Activation; Ovarian Neoplasms; Picibanil; Recombinant Proteins; T-Lymphocytes; Tumor Cells, Cultured

Human ovarian clear cell carcinoma cell line (transferrin (Tf)-non-producer), HAC 2, cells were adapted to grow in chemically defined synthetic medium when the cells were cultured with medium containing 10 micrograms/ml of insulin at least for 6 months. They synthesized and secreted constantly the 80 kDa protein immunologically similar to human serum Tf (15 +/- 12 ng/ml/10(7) cells/3 days). By sensitive lectin-affinity electrophoresis followed by antibody-affinity blotting technique, a concanavalin A weakly bound or unbound, lentil lectin, a strongly reactive abnormal band, which was rarely found in human serum Tf, was detectable in the Tf synthesized by HAC 2 cells (HACTf). These findings suggest that the HACTf may act as one of the autocrine growth factors and that this heterogeneity of HACTf for lectin affinity is ascribed to differences in the carbohydrate moiety of the Tf.

Topics: Adenocarcinoma; Chromatography, Affinity; Concanavalin A; Female; Humans; Immunoelectrophoresis; Lectins; Molecular Weight; Neoplasm Proteins; Ovarian Neoplasms; Plant Lectins; Precipitin Tests; Transferrin; Tumor Cells, Cultured

The placental alkaline phosphatase was purified by immunoaffinity chromatography from ovarian epithelial tumours to homogeneity. Up to 40% of the catalytical phosphatase activity in these tumours was derived from this placental type alkaline phosphatase (PLAP). The purified enzyme were similar to those of PLAP, whereas the PLAP-like isozyme was more heat-stable and resistant to 8 M urea than PLAP. The amino terminal sequence of the PLAP-like enzyme demonstrated heterogeneity at position three in the N-terminal end compared with PLAP. Phenyl-Sepharose affinity chromatography and different lectin chromatographies demonstrated the tumour-derived enzyme to be microheterogeneous, both with regard to concanavalin A binding and hydrophobicity properties.

Topics: Alkaline Phosphatase; Biomarkers, Tumor; Chromatography, Affinity; Concanavalin A; Cystadenoma; Electrophoresis; Female; GPI-Linked Proteins; Humans; Isoenzymes; Ovarian Neoplasms

In order to differentiate yolk sac-type alpha-fetoprotein (AFP) from hepatic-type AFP, 5 yolk sac tumors (YSTs) and 6 hepatocellular carcinomas (HCCs) were examined immunohistochemically by the peroxidase-antiperoxidase (PAP) method for AFP, and paradoxical concanavalin A (P-Con A) staining, which has been reported to detect glycoprotein including AFP. In all 5 YSTs, AFP was negative for P-Con A staining. On the other hand, AFP was strongly positive for the same staining in all 6 HCCs. A similar staining pattern for AFP was observed in human yolk sac endodermal cells and embryonal hepatocytes. Thus, it was clarified that yolk sac-type AFP was unable to bind with Con A, in contrast with hepatic-type AFP, on tissue sections. It was concluded that the PAP method for AFP and P-Con A staining might facilitate the immunohistochemical differentiation of these two types of AFP, and that it would be useful for clarifying the histogenesis of various AFP-secreting tumors.

Topics: alpha-Fetoproteins; Carcinoma, Hepatocellular; Concanavalin A; Female; Humans; Immunohistochemistry; Liver; Liver Neoplasms; Mesonephroma; Ovarian Neoplasms; Staining and Labeling; Yolk Sac

A sensitive new technique for lectin-affinity immunoelectrophoresis was applied to samples from 28 infants and children in order to distinguish the origin of elevated alpha-fetoprotein (AFP) in sera. This new immunoelectrophoresis was successfully performed within 24 hours in sera with AFP as small as 910 ng/mL. With combined use of concanavalin A (Con A) and lentil agglutinin (LCH) binding tests, AFPs were classified into three subtypes: benign hepatic condition type (six patients), hepatocellular carcinoma type (nine patients) and yolk sac type (12 patients). AFP was of hepatocellular carcinoma type in all seven patients with hepatoblastoma, and of benign hepatic condition type in six of seven patients with elevated AFP due to conditions such as hepatitis, biliary atresia, and normal newborn. The question as to whether AFP produced in "hepatoblastoma" is of benign hepatic condition type or hepatocellular carcinoma type was first answered by the information in this present report. The differentiation between yolk sac and general hepatic AFPs was completed with the Con A binding test.

Topics: Adolescent; alpha-Fetoproteins; Carcinoma, Hepatocellular; Concanavalin A; Female; Hepatitis; Humans; Immunoelectrophoresis; Infant; Lectins; Liver Neoplasms; Mesonephroma; Neoplasms; Ovarian Neoplasms; Plant Lectins

Profiles of concanavalin A (Con A) and lentil agglutinin (LCH) affinity immunoelectrophoresis were compared for serum alpha-fetoprotein (AFP) from patients with yolk sac tumors and carcinomas of the gastrointestinal tract, in order to find some correlations between peaks of AFP subfractions detectable by two different lectins, and to investigate whether or not it is possible to prove that the binding of AFP to LCH is weakened to some extent if a fucosylated sugar chain has, in addition, a bisect N-acetylglucosamine (GlcNAc) attached to the beta-linked mannose. The results obtained with our improved techniques tend to indicate that a Con A-reactive AFP subfraction (peak a) corresponds to an LCH strongly reactive AFP (peak A), while a Con A-nonreactive AFP (peak b) corresponds to an LCH weakly reactive AFP (peak B). the authors consider the present data sufficient to support the above explanation.

Topics: alpha-Fetoproteins; Concanavalin A; Dysgerminoma; Female; Humans; Immunoelectrophoresis; Lectins; Mesonephroma; Ovarian Neoplasms; Pancreatic Neoplasms; Plant Lectins; Stomach Neoplasms

A macrophage migration inhibition factor (OC-MIF) has been isolated from ascites fluid of ovarian cancer patients by affinity chromatography on L-fucose-Sepharose 6B, and characterized biochemically. OC-MIF activity was purified approximately 10 000-fold as compared to the starting material. It exhibits molecular heterogeneity with respect to net charge and molecular weights. Compared to it, purified and radioiodinated OC-MIF is fairly homogeneous and contains a major protein component with a molecular mass of about 45 kD, and two isoelectric points of 3.0-4.0 and about 5.0. Rabbits were immunized with the highly purified MIF material and an antiserum was prepared and was used to prepare immunoadsorbent beads. Beads made with anti-OC-MIF antiserum, but not with rabbit control serum, could remove specifically OC-MIF activity and showed weak reactivity towards Con A induced MIF. Using a radioimmunoassay (RIA) anti-OC-MIF antiserum reacts with OC-MIF and also with Con A induced MIF. This antigenic relationship between conventional MIF and OC-MIF and common biochemical properties suggest that the two mediator substances are very similar and may, perhaps, be identical. Furthermore, the possibility to determine various MIF activities by means of RIA was investigated.

Topics: Antibodies; Ascites; Chromatography, Affinity; Chromatography, Gel; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Female; Humans; Immunosorbent Techniques; Isoelectric Focusing; Macrophage Migration-Inhibitory Factors; Macrophages; Ovarian Neoplasms; Radioimmunoassay

Using a modified method of Con A, LCH or PHA-E affinity crossed-line immunoelectrophoresis, we studied AFP subfractions in 78 sera including 58 from patients with primary hepatoma, 11 from patients with hepatic metastasis of gastric cancer and 9 from patients with germ cell tumors of the gonads (yolk sac tumor, immature solid teratoma or mature solid teratoma). It was found that AFP in primary hepatoma, metastatic hepatoma or germ cell tumors of the gonads were differently glycosylated, and different patterns of AFP subfractions identified by Con A, LCH or PHA-E affinity crossed-line immunoelectrophoresis facilitated a differential diagnosis of such AFP related malignancies.

Topics: alpha-Fetoproteins; Carcinoma, Hepatocellular; Concanavalin A; Female; Humans; Immunoelectrophoresis; Immunoelectrophoresis, Two-Dimensional; Lectins; Liver Neoplasms; Male; Mesonephroma; Neoplasms, Germ Cell and Embryonal; Ovarian Neoplasms; Teratoma; Testicular Neoplasms

We used affinity chromatography on concanavalin A Sepharose to study the serum alpha-fetoprotein of 10 patients with histologically proven germ-cell tumors and 12 patients with primary liver cancer. Less than 50% of the fetoprotein from germ-cell tumors bound to concanavalin A, as compared with more than 80% of the alpha-fetoprotein from primary liver cancers.

Topics: Adult; Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Chromatography, Affinity; Concanavalin A; Female; Humans; Liver Neoplasms; Male; Mesonephroma; Middle Aged; Ovarian Neoplasms; Sepharose; Testicular Neoplasms

We have tested 74 teratogenic and 28 nonteratogenic agents in a recently developed in vitro teratogen assay system. The assay identifies teratogens by their ability to inhibit attachment of ascites tumor cells to plastic surfaces coated with concanavalin A. There is a qualitative agreement between in vivo animal data and in vitro activity for 81 of the 102 agents (79%). Quantitative analysis shows a highly significant correlation coefficient of 0.69 between the inhibitory in vitro dose and the lowest reported teratogenic dose for 54 of the 60 inhibitory teratogens. The doses analyzed ranged over 5 orders of magnitude. We interpret these results to mean that attachment inhibition in concert with other, complementary, in vitro assay systems can become a useful method for the assessment of the teratogenic potential of environmental agents.

Topics: Animals; Cell Adhesion; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Drug; Female; Mice; Ovarian Neoplasms; Teratogens

Topics: Animals; Cell Adhesion; Cells, Cultured; Concanavalin A; Female; Mice; Mice, Inbred C3H; Microsomes, Liver; Neoplasms, Experimental; Ovarian Neoplasms; Teratogens; Thalidomide

In view of the reported disagreement in the physicochemical properties of ovarian carcinoembryonic antigen (CEA), this study was undertaken to compare the properties of CEA obtained from extracts of ovarian tumour tissue, ascitic fluid and cyst fluid. On the basis of molecular weight estimation and binding properties with Concanavalin A and wheat germ lectin, ovarian CEA from these three sources appeared similar, and also possessed similar properties to those of colonic CEA. On isoelectric focusing, however, it was found that the isoelectric point of CEA from tumour tissue and cyst fluid differed from that from ascitic fluid. It is most likely that this is due to a loss of sialic acid from the CEA released into ascitic fluid.

Topics: Ascites; Carcinoembryonic Antigen; Concanavalin A; Cystadenocarcinoma; Female; Humans; Ovarian Cysts; Ovarian Neoplasms; Radioimmunoassay

The HPRS line 1 lymphoblastoid cells derived from an ovarian lymphoma induced by Marek's disease virus in chickens displayed increased agglutinability and haemadsorption when incubated with Con A. The increase was proportional to the concentration of Con A used. The same Con A-untreated cells and normal chicken lymphocytes incubated with Con A exhibited no increase in agglutinability and haemadsorption, however.

Topics: Agglutination; Animals; Cell Line; Cell Membrane; Chickens; Concanavalin A; Erythrocytes; Female; Hemadsorption; Herpesvirus 2, Gallid; Humans; Lymphoma; Ovarian Neoplasms; Receptors, Concanavalin A

Human AFP purified from fetal serum and amniotic fluid was separated into three different variants by chromatography on concanavalin A insolubilized on Sepharose (Con A--Sepharose). The three variants were indistinguishable in immunodiffusion and radioimmunoassay. Sera from patients with yolk-sac tumor and amniotic fluid from early pregnancy were found to contain a high proportion (15-45%) of AFP which does not bind to Con A, while AFP in fetal and newborn sera, and in amniotic fluid from late pregnancy, contained less (2-6%) of this variant. The use of a large excess of Con A--Sepharose and the fact that the non-bound AFP consistently eluted as non-bound in rechromatography showed that this AFP is non-reactive with Con A. Fractionation of radiolabelled AFP from cord serum in a mixture with amniotic fluid verified the difference in the amount of the Con-A nonreactive variant in AFP from these two sources. These results suggest that AFP synthesized by the yolk-sac tissue and by the liver are glycosylated differently. The variant may prove to be diagnostically useful.

Topics: alpha-Fetoproteins; Amniotic Fluid; Carbohydrates; Chromatography, Affinity; Concanavalin A; Female; Fetal Blood; Humans; Immunodiffusion; Infant, Newborn; Liver; Liver Neoplasms; Mesonephroma; Ovarian Neoplasms; Pregnancy; Pregnancy Trimester, Second; Pregnancy Trimester, Third; Radioimmunoassay; Teratoma

Human ovarian tumor-associated antigen (TAA) has been purified from ovarian tumor tissue by affinity chromatography on concanavallin A-Sepharose and three different gamma globulin-Sepharose columns. The resulting ovarian TAA appears to be contaminated by one normal antigen or family of antigens. Rabbit antiserum prepared against this purified ovarian TAA (antiserum 404) was coupled to CNBr-activated Sepharose 4B. This coupled Sepharose was added to fractionated serum from ovarian cancer patients with Stage III and IV malignancy. Bound protein was eluted with 0.2M glycine buffer and tested against antiserum 404. The bound protein contained TAA identical to the TAA isolated from ovarian tumor tissue.

Topics: Animals; Antigens, Neoplasm; Chromatography, Affinity; Concanavalin A; Cyanogen Bromide; Female; gamma-Globulins; Humans; Immune Sera; Immunodiffusion; Immunoelectrophoresis; Mannose; Ovarian Neoplasms; Ovary; Rabbits; Sepharose

This study was designed to answer the question, do molecules with carcinoembryonic antigen (CEA) activity from colon, breast, and ovarian cancer differ? Extracts of two breast and three ovarian cancers with CEA activity were compared to three colon cancer CEA preparations and to the related antigen, colon carcinoma antigen-III, in terms of lectin- and antiserum-binding properties. With the use of Farr-type radioimmunoassays with the lectins, concanavalin A and wheat germ agglutinin, the iodinated colon CEA and CEA-like preparations from breast and ovarian cancer all showed distinctly different patterns of binding. Specificity of binding was confirmed by inhibition studies with the relevant monosaccharides. Similarly, with antisera prepared against colon CEA, colon carcinoma antigen-III, or breast CEA, it was shown that, although all preparations shared some antigens, unique antigenic determinants were also present on all preparations. These data are consistent with the concept of a series of closely related CEA and CEA-like molecules with distinct characteristics for each tissue source of CEA.

Topics: Antibodies, Neoplasm; Antibody Specificity; Antigens, Neoplasm; Binding Sites; Binding Sites, Antibody; Binding, Competitive; Breast Neoplasms; Carcinoembryonic Antigen; Chromatography; Colonic Neoplasms; Concanavalin A; Epitopes; Female; Humans; Immunosorbent Techniques; Lectins; Ovarian Neoplasms; Radioimmunoassay

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: Agglutination; Animals; Antigens, Viral; Cattle; Cell Line; Cell Membrane; Chick Embryo; Chickens; Chromosome Aberrations; Clone Cells; Complement Fixation Tests; Concanavalin A; Culture Media; Ducks; Female; Fluorescent Antibody Technique; Herpesviridae; Marek Disease; Microscopy, Electron; Ovarian Neoplasms; Spleen; Splenic Neoplasms; Viral Proteins