concanavalin-a and Colonic-Neoplasms

Fluorescence resonance energy transfer (FRET) from a donor-labelled molecule to an acceptor-labelled molecule is a useful, proximity-based fluorescence tool to discriminate molecular states on the surface and in the interior of cells. Most microscope-based determinations of FRET yield only a single value, the interpretation of which is necessarily model-dependent. In this paper we demonstrate two new measurements of FRET heterogeneity using selective donor photobleaching in combination with synchronous donor/acceptor detection based on either (1) full kinetic analysis of donor-detected and acceptor-detected donor photobleaching or (2) a simple time-based ratiometric approach. We apply the new methods to study the cell surface distribution of concanavalin A yielding estimates of FRET and non-FRET population distributions, as well as FRET efficiencies within the FRET populations.

Topics: Animals; Cell Line, Tumor; Cell Membrane; Colonic Neoplasms; Concanavalin A; Fluorescence Recovery After Photobleaching; Fluorescence Resonance Energy Transfer; Mice; Microscopy, Fluorescence, Multiphoton; Protein Interaction Mapping

Expression of decay-accelerating factor (DAF, CD55), a complement-regulatory glycoprotein, is enhanced in colorectal-cancer (CC) cells and colonic epithelium in ulcerative colitis (UC), and stools from these patients contain increased amounts of DAF. Carbohydrate chains of glycoproteins are often altered during malignant transformation or inflammation. In this study, we investigated whether DAF molecules in patients with CC and those with UC differ with respect to oligosaccharide side chains. We analyzed DAF in stools and homogenates of colonic-tissue specimens obtained from patients with CC or UC using solid-phase enzyme-linked assay and Western blotting for reactivity with the lectins Ulex europaeus agglutinin I (UEA-I), wheat-germ agglutinin, peanut agglutinin, and concanavalin A. UEA-I bound to DAF in stools from patients with UC but not in that from the stools of CC patients, as demonstrated on the solid-phase enzyme-linked assay (P <.05, Mann-Whitney U test) and Western blotting. Binding of UEA-I was specifically inhibited by the addition of fucose. The difference in UEA-I reactivity with DAF was observed also in colonic-tissue homogenates from patients with UC and those with CC. DAF expressed in the mucosa and excreted into the stools of UC patients is different from that expressed in CC with regard to UEA-I reactivity. Future studies should be directed toward determining whether a qualitatively unique isoform of DAF is present, of which sugar chains are specific to CC in UC patients.

Topics: Adenocarcinoma; Adult; Aged; CD55 Antigens; Colitis, Ulcerative; Colon; Colonic Neoplasms; Colorectal Neoplasms; Concanavalin A; Feces; Female; Humans; Intestinal Mucosa; Kinetics; Lectins; Male; Middle Aged; Neoplasm Staging; Plant Lectins; Rectal Neoplasms

Concanavalin A (ConA), directly injected into mice, induces T cell-mediated liver injury. However, it remains unclear whether ConA injection can activate innate immune cells, including natural killer (NK) cells and natural killer T (NKT) cells, both of which exist abundantly in the liver. Here we report that ConA injection stimulated interferon (IFN)-gamma production from liver NKT cells as early as 2 hours after injection and augmented YAC-1 cytotoxicity of liver NK cells. ConA-induced NK cell activation required other types of immune cells and critically depended on IFN-gamma. Because a nonhepatotoxic low dose of ConA was capable of fully activating both NKT cells and NK cells, we next addressed the possibility of ConA injection displaying an antitumor effect in the liver without liver injury. A nonhepatotoxic low-dose ConA injection augmented the cytotoxicity of liver NK cells against Colon-26 colon cancer cells and suppressed hepatic metastasis of Colon-26 cells in a NK cell- and IFN-gamma-dependent manner. In conclusion, a nonhepatotoxic low dose of ConA might serve as an immunomodulator that can preferentially activate the innate immune cells to induce an antitumor effect against metastatic liver tumor.

Topics: Animals; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Colonic Neoplasms; Concanavalin A; Cytotoxicity, Immunologic; Dose-Response Relationship, Drug; Female; Immunologic Factors; Injections, Intravenous; Interferon-gamma; Killer Cells, Natural; Liver; Liver Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; T-Lymphocytes

Human colon carcinoma cell fucosyltransferase (FT) in contrast to the FTs of several human cancer cell lines, utilized GlcNAcbeta1,4GlcNAcbeta-O-Bn as an acceptor, the product being resistant to alpha1,6-L-Fucosidase and its formation being completely inhibited by LacNAc Type 2 acceptors. Further, this enzyme was twofold active towards the asialo agalacto glycopeptide as compared to the parent asialoglycopeptide. Only 60% of the GlcNAc moieties were released from [14C]fucosylated asialo agalacto triantennary glycopeptide by jack bean beta-N-acetylhexosaminidase. These alpha1,3-L-fucosylating activities on multiterminal GlcNAc residues and chitobiose were further examined by characterizing the products arising from fetuin triantennary and bovine IgG diantennary glycopeptides and their exoglycosidase-modified derivatives using lectin affinity chromatography. Utilization of [14C]fucosylated glycopeptides with cloned FTs indicated that Lens culinaris lectin and Aleuria aurantia lectin (AAL) required, respectively, the diantennary backbone and the chitobiose core alpha1,6-fucosyl residue for binding. The outer core alpha1,3- but not the alpha-1,2-fucosyl residues decreased the binding affinity of AAL. The AAL-binding fraction from [14C]fucosylated asialo fetuin, using colon carcinoma cell extract, contained 60% Endo F/PNGaseF resistant chains. Similarly AAL-binding species from [14C]fucosylated TFA-treated bovine IgG using colon carcinoma cell extract showed significant resistance to endo F/PNGaseF. However, no such resistance was found with the corresponding AAL non- and weak-binding species. Thus colon carcinoma cells have the capacity to fucosylate the chitobiose core in glycoproteins, and this alpha1,3-L-fucosylation is apparently responsible for the AAL binding of glycoproteins. A cloned FT VI was found to be very similar to this enzyme in acceptor substrate specificities. The colon cancer cell FT thus exhibits four catalytic roles, i.e., alpha1,3-L-fucosylation of: (a) Galbeta1,4GlcNAcbeta-; (b) multiterminal GlcNAc units in complex type chain; (c) the inner core chitobiose of glycopeptides and glycoproteins; and (d) the nonreducing terminal chiotobiose unit.

Topics: Animals; Cattle; Chromatography, Affinity; Cloning, Molecular; Colonic Neoplasms; Concanavalin A; Disaccharides; Fucosyltransferases; Glycopeptides; Humans; Immunoglobulin G; Lectins; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; Plant Lectins; Substrate Specificity; Tumor Cells, Cultured

Direct intratumor injection of a disabled infectious single cycle HSV-2 virus encoding the murine GM-CSF gene (DISC/mGM-CSF) into established murine colon carcinoma CT26 tumors induced a significant delay in tumor growth and complete tumor regression in up to 70% of animals. Pre-existing immunity to HSV did not reduce the therapeutic efficacy of DISC/mGM-CSF, and, when administered in combination with syngeneic dendritic cells, further decreased tumor growth and increased the incidence of complete tumor regression. Direct intratumor injection of DISC/mGM-CSF also inhibited the growth of CT26 tumor cells implanted on the contralateral flank or seeded into the lungs following i.v. injection of tumor cells (experimental lung metastasis). Proliferation of splenocytes in response to Con A was impaired in progressor and tumor-bearer, but not regressor, mice. A potent tumor-specific CTL response was generated from splenocytes of all mice with regressing, but not progressing tumors following in vitro peptide stimulation; this response was specific for the gp70 AH-1 peptide SPSYVYHQF and correlated with IFN-gamma, but not IL-4 cytokine production. Depletion of CD8(+) T cells from regressor splenocytes before in vitro stimulation with the relevant peptide abolished their cytolytic activity, while depletion of CD4(+) T cells only partially inhibited CTL generation. Tumor regression induced by DISC/mGM-CSF virus immunotherapy provides a unique model for evaluating the immune mechanism(s) involved in tumor rejection, upon which tumor immunotherapy regimes may be based.

Topics: Animals; Colonic Neoplasms; Combined Modality Therapy; Concanavalin A; Cytokines; Cytotoxicity, Immunologic; Dendritic Cells; Epitopes; Female; Genetic Vectors; Granulocyte-Macrophage Colony-Stimulating Factor; Herpes Simplex; Herpesvirus 1, Human; Immunity, Active; Injections, Subcutaneous; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Spleen; T-Lymphocytes, Cytotoxic; Virus Activation

Tissue characterisation by fluorescence imaging, using exogenous fluorophores, is a promising method for cancer detection. Histochemical alterations in the composition of mucins, when neoplastic transformations occur, could be exploited to derive more selective fluoroprobes indicative of early malignant transformation. The aim of this work was to develop and examine tumour selective fluoroprobes for colon cancer diagnosis, as well as to determine the morphological components where selective dye accumulation has occurred. Two novel fluoroprobes: rhodamine B-L-leucine amide and rhodamine B-phenylboronic acid were synthesised and examined together with Mayer's mucicarmine, alexa 350-wheat germ agglutinin (WGA) and tetramethyl rhodamine-concanavalin A (ConA). Fluorescence microscopy studies were performed with deparaffinised human colon sections, using an epifluorescence microscope equipped with a colour CCD camera. The intense accumulation of the novel fluoroprobes was localised in the amorphous material in the lumen of neoplastic crypts. To gain insight into the localisation patterns, mucicarmine, alexa 350-WGA and tetramethyl rhodamine-ConA were used. Alexa 350-WGA reacted primarily with mucin secreted in the malignant crypt lumen suggesting that this material is rich in sialic acid and N-acetylglucosaminyl residues. These derivatives clearly and consistently distinguished non-neoplastic from neoplastic human colon tissue sections. The intense accumulation at the altered mucins indicates that they could be used as fluoroprobes of biochemical alterations for carcinoma detection.

Topics: Carmine; Colonic Neoplasms; Coloring Agents; Concanavalin A; Humans; Indicators and Reagents; Microscopy, Fluorescence; Rhodamines; Wheat Germ Agglutinins

Natural killer (NK) cells are non-T, non-B cell lymphocytes that lyse a variety of tumor and virus-infected cells. In this study, we demonstrated that phytohemagglutinin (PHA) rendered resistant autologous T cells extremely sensitive to natural-killer(NK)-cell-mediated lysis. The sensitization was very rapid and concentration-dependent (0.01-1 microg/ml); 62% and 95% of autologous T cells were lysed by interleukin-2-activated NK cells 5 min and 18 h respectively after treatment with PHA (1 microg/ml). The maximal decrease in the level of MHC class I molecules observed on T cells was 22%. Induction of susceptibility to NK-mediated lysis was correlated with the expression of activation markers on T cells treated for relatively long intervals (more than 18 h) with high concentrations of PHA (more than 0.1 microg/ml). Sensitization of T cells required RNA and protein synthesis, although DNA synthesis was not essential. We propose that this unique system is suitable for studying the mechanisms involved in recognition and killing of target cells by NK cells.

Topics: Adenocarcinoma; Colonic Neoplasms; Concanavalin A; Cycloheximide; Cytotoxicity, Immunologic; Dactinomycin; DNA Replication; Humans; Interleukin-2; K562 Cells; Killer Cells, Natural; Lymphocyte Activation; Mitomycin; Nucleic Acid Synthesis Inhibitors; Phytohemagglutinins; Protein Biosynthesis; Protein Synthesis Inhibitors; RNA; T-Lymphocyte Subsets; Tumor Cells, Cultured

An in vitro cytotoxic screening of extracts of Rhaphidophora korthalsii indicated cytotoxicity in the ether fraction. ED50 values of the extract against P388, Molt 4, KB and SW 620 were 12, 14, 8 and 13 micrograms/ml, respectively. The extract was relatively more toxic on P388 and Molt 4 cell lines at concentrations of 50 micrograms/ml and 100 micrograms/ml. Screening with mouse splenocytes showed that the hot water extract had splenocytes stimulating activity.

Topics: Adenocarcinoma; Adjuvants, Immunologic; Animals; Antineoplastic Agents, Phytogenic; Carcinoma, Squamous Cell; Cell Division; Colonic Neoplasms; Coloring Agents; Concanavalin A; Dose-Response Relationship, Drug; Female; Humans; Leukemia; Leukemia P388; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Nasopharyngeal Neoplasms; Plant Extracts; Plant Lectins; Plants, Medicinal; Spleen; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured; Water

In human intestinal malignancy, alterations occur in the expression of mucins defined by the MUC-2 gene. These changes include the unmasking of epitopes in the mucin protein core. In order to probe these modifications associated with mucins of the malignant phenotype, a monoclonal antibody (MAb) was developed against synthetic peptide with a sequence based upon that of the protein core of the MUC-2 mucin. The antibody (designated 996) was shown to recognize a high-molecular-weight glycoprotein from colonic carcinoma tissue. The material reacted uniformly with Concanavalin A but variably with other lectins, indicating heterogeneity in the associated oligosaccharide side chains. The protein core was accessible both to 996 antibody binding and to degradation with proteases. Immunization with the affinity-purified mucin-like material elicited antibodies reactive with both the immunogen and the synthetic peptides, confirming the immunogenic character of protein-core determinants. Epitope mapping studies, using synthetic peptides in solution and synthetic peptides tethered to the heads of plastic pins, indicated that the minimum epitope for the 996 antibody is a tetramer of T G T Q. Antibody interaction with the glutamine (Q) residue was determined to be of major importance in the antigen-antibody reaction. The findings illustrate the characterization of an anti-peptide antibody which may be used to probe alterations in MUC-2 mucin expression associated with human intestinal malignant disease.

Topics: Adsorption; Amino Acid Sequence; Antibodies, Monoclonal; Antibody Specificity; Chymotrypsin; Colonic Neoplasms; Concanavalin A; Epitopes; Glutamine; Humans; Immunoblotting; Molecular Sequence Data; Mucin-2; Mucins; Neoplasm Proteins; Trypsin

A molecular form of CEA non-binding Con A (CEAnC) was isolated from colon adenocarcinoma metastases in liver as a fraction of CEA having no affinity to Con-A-Sepharose. CEAnC was shown to be immunochemically identical to CEA, but to differ substantially with regard to the amino acid and sugar composition, and structure of the sugar moiety, possibly containing non only N-, but also O-glycosyl carbohydrate chains. The antigens studied were also found to possess different spatial structures. The differences between CEA and CEAnC suggest CEAnC to be a new molecular form of CEA.

Topics: Adenocarcinoma; Amino Acids; Carbohydrate Sequence; Carcinoembryonic Antigen; Chromatography, Gel; Chromatography, Ion Exchange; Circular Dichroism; Colonic Neoplasms; Concanavalin A; Humans; Immunochemistry; Immunodiffusion; Liver Neoplasms; Molecular Sequence Data

This study was undertaken to determine whether infusion of a unique ZCE/CHA bifunctional antibody (BFA, 5-40 mg) could alter the composition and functions of peripheral blood leucocytes in 18 patients with colon cancer. The BFA is made by combining chemically the Fab' fragments of two murine monoclonal antibodies. One fragment (ZCE 025) binds to the carcino-embryonic antigen (CEA) and the other (CHA 225) to an epitope, present on an 111In-benzyl EDTA analog of bleomycin (BLEDTA IV) and on 111In-hydroxy-ethyl-thiourea benzyl EDTA (EOTUBE). The radiolabelled epitope (111In-BLEDTA IV or 111In-EOTUBE) was given 4 days after prelocalization with BFA. Peripheral blood samples were tested before BFA infusion, at the end of infusion (1 h later), and at 4 and 7 days post-infusion. A 50% or greater suppression in lymphocyte responsiveness to phytohaemagglutinin (PHA) and concanavalin A (Con A) was seen in 13 out of 18 and 12 out of 18 subjects, respectively, at some time after BFA infusion; this was especially evident in those patients with pre-infusion stimulation indices of greater than 50 (PHA) and/or greater than 10 (Con A). In contrast, natural killer (NK) cell cytotoxicity and oxygen radical production increased in five out of 15 and in seven out of 18 subjects, respectively. Little or no change was observed in CD3, CD4, CD8, CD16, and CD19 markers on lymphocyte subpopulations as determined by flow cytometry. These data suggest that significant changes in mitogen-induced lymphoproliferation. NK cell cytotoxicity, and oxygen radical production can occur in a substantial proportion of cancer patients after infusion of the ZCE/CHA bifunctional antibody system. The immunomodulation was unrelated to initial BFA dose, dose of BFA as a carrier, or to subsequent infusion of either form of the 111In epitope. The clinical significance of these phenomena, if any, remains to be determined.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Antibodies, Monoclonal; Antigens, CD; Colonic Neoplasms; Concanavalin A; Cytotoxicity, Immunologic; Dose-Response Relationship, Drug; Female; Flow Cytometry; Humans; Immunotherapy; Killer Cells, Natural; Leukocytes; Lymphocyte Activation; Male; Middle Aged; Phytohemagglutinins; Time Factors

Peripheral blood samples from six cancer patients (five colon cancer, one lung cancer) and six healthy volunteers were tested in vitro for oxygen radical production by phagocytic cells and in assays of mitogen-induced lymphoblastogenesis at physiologic and pharmacologic concentrations of pyridoxine (PN, 1.8-96 nmol/ml) or pyridoxal (PL, 0.08-90 nmol/ml). Plasma levels of pyridoxal-5'-phosphate (PLP), 4-pyridoxic acid (4PA), pyridoxamine phosphate (PMP), and PL were also determined. Phagocytic cells from three patients showed significantly increased capacity for oxygen radical production when incubated in PL-, but not PN-supplemented media. Oxygen burst capacity of cells from healthy subjects was significantly enhanced by PN-, but not PL-enriched media. Lymphocyte responsiveness to phytohemagglutinin or pokeweed mitogen (PWM) stimulation showed a modest increase in cell activation in three patients as the concentration of PN was increased; with concanavalin A, two showed enhanced responsiveness. On the other hand, PL-supplementation resulted in greater cell proliferation only with PWM. The cancer patients had significantly lower plasma PLP, 4PA, and PMP levels when compared with the healthy volunteers. These data indicate that in the cancer patients and in a majority of the healthy volunteers, vitamin B-6 status was marginal or deficient and suggest that increasing PN or PL in vivo levels may augment functions related to cell-mediated immunity.

Topics: Adult; Aged; Colonic Neoplasms; Concanavalin A; Female; Free Radicals; Humans; Leukocytes; Lung Neoplasms; Lymphocyte Activation; Male; Middle Aged; Oxygen; Phytohemagglutinins; Pokeweed Mitogens; Pyridoxal; Pyridoxine

We studied the effects on platelet function of cells isolated from freshly dissociated human tumor tissues (11 breast carcinomas, 9 colon carcinomas and 1 lymph node metastasis from melanoma) obtained at surgery as compared with cultured human tumor cells: namely, human melanoma 1402 cell line derived from a primary tumor and two lines derived from lymph node metastases (ME 7110/2 and Me 665/1) as well as a human hepatoma cell line (Hep G2). The three melanoma cell lines activated platelets by producing ADP, as evidenced by the inhibitory effect of apyrase and by the direct measurement of the agonist in the supernatants of tumor cell suspensions; this production was much greater by the cells derived from metastases than by the cells derived from the primary tumor. On the other hand, aggregation induced by Hep G2 hepatoma cells was unaffected by apyrase and was inhibited by hirudin or concanavalin A, suggesting that the cells aggregate platelets by producing thrombin, probably through tissue factor activity of the cells themselves. Cells isolated from 16 of the 21 human tumor tissues possessed a potent platelet-aggregating effect, which was not inhibited by apyrase, hirudin or concanavalin A, but was virtually abolished by the cysteine protease inhibitors iodoacetic acid or p-hydroxymercuri-phenylsulfonate. Collectively, our data demonstrate that cells isolated from freshly dissociated tumor tissues activate platelets through tumor-associated cysteine proteinases rather than by the ADP- or thrombin-dependent mechanisms characteristic of cultured human tumor cell lines.

Topics: Adenosine Diphosphate; Adult; Apyrase; Blood Platelets; Breast Neoplasms; Cell Communication; Cell Separation; Colonic Neoplasms; Concanavalin A; Cysteine Endopeptidases; Cysteine Proteinase Inhibitors; Hirudins; Humans; Melanoma; Platelet Aggregation; Tumor Cells, Cultured

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

FITC-Con A staining as a rapid diagnostic method for tumor cells was applied to the tumors smeared on glass slide and section specimens to evaluate the reactivity with FITC-Con A. Good staining results were obtained in smear specimens with clear fluorescence on the membrane of tumor cells. Con A and LCH lectins bound well with tumor cells to produce strong fluorescence in comparison with PEA and DBA. It indicates that tumor cells expressed dominantly the receptors of alpha-D-glucose and alpha-D-mannose sugar chain on the membrane of tumor cells. From these results it was concluded that FITC-Con A staining method applied to smear specimens is more advantageous in the rapidity and the simplicity for tumor cell diagnosis than section specimen method.

Topics: Animals; Colonic Neoplasms; Concanavalin A; Fluorescein-5-isothiocyanate; Fluoresceins; Mice; Mice, Inbred BALB C; Microtomy; Specimen Handling; Staining and Labeling; Tumor Cells, Cultured

Topics: Adenocarcinoma; Animals; Colonic Neoplasms; Concanavalin A; Cyclic AMP; Heat-Shock Proteins; Humans; Hyperthermia, Induced; Lysine; Membrane Glycoproteins; Neoplasm Proteins; Predictive Value of Tests; Protein Biosynthesis

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

To establish a method for evaluation of immunological parameters in small blood samples, a whole blood technique was developed for the estimation of mitogen- or antigen-induced proliferation. Studies regarding cellular immunity in patients with colon cancer were done with 108 patients in all tumor stages, aged 32 to 80 years. They were studied before surgery and 10 days after operation. A group of 35 patients were further tested 3 months after surgical treatment. In patients with colon cancer the proliferative responses of peripheral blood lymphocytes to mitogens were significantly lower in comparison to healthy controls. These results were found when the response to concanavalin A, phytohemagglutinin, OKT 3, and pokeweed mitogen were analyzed preoperatively and 10 days postoperatively. There was no relation to the stage of disease. The marked reduction of mitogen responses was followed by a gradual return toward normal values 3 months after surgical resection of neoplastic growth in 80% of the patients. Our studies indicate that the defects were largely restored when testing was performed 3 months after operation. Using this result, it will be possible to perform long-term studies in order to establish if there is a correlation between the return to normal immune reactivity and the survival of individual patients.

Topics: Adult; Aged; Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; Colonic Neoplasms; Concanavalin A; Humans; Lymphocyte Activation; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; Time Factors; Tuberculin

Topics: 1,2-Dimethylhydrazine; Adenocarcinoma; Animals; Colonic Neoplasms; Concanavalin A; Dimethylhydrazines; Female; Glycoproteins; Lectins; Mice; Mice, Inbred ICR

Topics: 1,2-Dimethylhydrazine; Animals; Colon; Colonic Neoplasms; Concanavalin A; Dimethylhydrazines; Female; Glucose; Mannose; Mice

A lectin reactivity specific to human bowel carcinoma is reported. Twenty-six cases of carcinoma of the large intestine were examined. Normal as well as transitional mucosa and carcinoma tissues were removed from surgical specimens, and paraffin sections were stained with a battery of histochemical methods to characterize glycoconjugates, including high iron diamine-Alcian blue pH 2.5, modified PAS reaction to detect various sialic acids, paradoxical concanavalin A (Con A) staining, and stainings with 10 species of lectins labeled with horseradish peroxidase (HRP). Among the techniques employed, only Griffonia simplicifolia agglutinin-II (GS-II, specific to glucosamine)-HRP staining revealed highly selective affinity to the carcinoma tissues; the apical surface of the carcinoma cells stained most intensely. GS-II reactivity of the cells persisted after prior periodate oxidation, but was significantly enhanced by neuraminidase digestion. Comparison with two other lectin stainings with the same sugar specificity, viz. paradoxical concanavalin A staining and wheat germ agglutinin (WGA)-HRP staining, showed that the GS-II reactive sites lacked class III Con A reactivity but were possibly included in WGA reactive sites. The GS-II-HRP staining should be helpful in the identification of carcinoma tissue and for analysis of carcinoma-associated antigens.

Topics: Antigens, Neoplasm; Carcinoma; Colonic Neoplasms; Concanavalin A; Humans; Intestinal Mucosa; Lectins; Mucins; Neuraminidase; Oxidation-Reduction; Periodic Acid; Plant Lectins; Staining and Labeling; Wheat Germ Agglutinins

The nature of the inhibitory activity of sera of patients with metastatic cancer on in vitro immunoassays remains unclear. Serum glycoprotein levels in cancer patients show a reasonable correlation with the clinical status, especially with progressive metastatic disease. Glycoproteins like acute phase reactants have been connected with immunosuppressive activity in cancer patients' sera. In this study, we examined the influence of glycoprotein fractions of normal and cancer sera, separated by Con A immunoadsorption, on the mixed lymphocyte culture as a reference system for suppressive activity. Glycoprotein rich fractions with the utmost recovery of the acute phase reactants inhibited the mixed lymphocyte culture in a dose-dependent manner. This effect was more pronounced in patients sera as compared to control sera. But there is evidence of additional blocking activity in the glycoprotein poor serum fraction, indicating blocking factors still to be identified.

Topics: Blood Proteins; Breast Neoplasms; Chromatography, Affinity; Colonic Neoplasms; Concanavalin A; Dose-Response Relationship, Immunologic; Electrophoresis, Polyacrylamide Gel; Glycoproteins; Humans; Immune Tolerance; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Neoplasm Proteins; Neoplasms; Rectal Neoplasms; Suppressor Factors, Immunologic

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

In the adenocarcinoma cell line HT-29 receptor-bound insulin is substrate for a proteolytic process leading to the release of about half of the cell-associated [125I]monoiodoinsulin in the form of [125I]iodide and [125I]monoiodotyrosine. Classical lysosomal inhibitors (NH+4, methylamine, leupeptin) did not inhibit this proteolysis. Inhibitors of membrane traffic (chloroquine and monensin) and of metabolism (CN-) inhibited the fractional receptor-mediated degradation. The former led to an increased cell-associated 125I activity whereas the latter reduced the uptake. Sulphydryl reagents inhibited the receptor-mediated degradation. The data are not compatible with a quantitatively major role of lysosomes in the receptor-mediated insulin degradation. However, since the process requires energy it is suggested that the receptor-mediated degradation takes place in vesicles other than secondary lysosomes. The responsible enzyme(s) may belong to the thiol group of proteases. Both insulin and the insulin receptor are internalized as a consequence of incubation of HT-29 cells with insulin.

Topics: Adenocarcinoma; Adipose Tissue; Animals; Cell Line; Colonic Neoplasms; Concanavalin A; Fibroblasts; Humans; Hydrogen-Ion Concentration; Insulin; Iodine Radioisotopes; Lymphocytes; Rats; Receptor, Insulin; Sulfhydryl Reagents; Temperature

The immune regulation of phytohemagglutinin (PHA) and concanavalin A (Con A) mitogen responses by prostaglandin (PG)-producing suppressor monocytes was examined in 57 patients with colorectal cancer and 55 normal individuals. The blood lymphocyte responses to either PHA or Con A were significantly depressed in 74% of patients compared to normal controls. The mean PHA response for the patients was significantly lower than that for controls (17,649 versus 25,549 cpm, P = 0.02), while the mean Con A response for the patients was also depressed but not as significantly (13,551 versus 18,623 cpm, P = 0.09). The depression of immune competence was greatest in older patients and those with metastatic disease. The addition of indomethacin (1 microgram/ml) to cell cultures of both patients and normal individuals enhanced the mitogen response, suggesting that PGE-producing suppressor cells were operative in both groups. Among the patient group, however, a differential modulation of the immune response by indomethacin was observed. Thus, the addition of indomethacin restored the PHA response in patients almost to normal levels, while the Con A increase was less pronounced. Even after indomethacin treatment, the Con A proliferative response by lymphocytes was significantly depressed in patients as compared to controls (P = 0.002). To prove that indomethacin was blocking excessive PG production by suppressor monocytes in colon cancer patients, we directly measured PGE2 production by peripheral blood mononuclear cells (PBMCs) using a radioimmunoassay. PBMCs from the patients produced significantly greater amounts of PGE2 compared to controls (10.1 versus 5.1 ng/ml, P = 0.0001). This comparison was still significant after adjustment for age and sex. The increased PGE2 production appeared to be selective, since the levels of two other arachidonic acid metabolites, PGF1 alpha and thromboxane B2, were the same or less than control levels. PG-mediated immune suppression of mitogenesis thus appears to be abnormally increased in colon cancer patients, particularly for the PHA response. This abnormality was partially corrected in vitro by incubation of the PBMCs with indomethacin, a prostaglandin synthetase inhibitor.

Topics: Adult; Aged; Colonic Neoplasms; Concanavalin A; Dinoprostone; Female; Humans; Immunity, Cellular; Indomethacin; Male; Middle Aged; Mitosis; Phytohemagglutinins; Prostaglandins E; Prostaglandins F; Radioimmunoassay; T-Lymphocytes, Regulatory; Thromboxane B2

Two variants of gamma-glutamyltransferase were demonstrated in colorectal carcinomas. When compared to the enzyme in normal large bowel mucosa, one of the variants showed reduced affinity to concanavalin A and considerable charge heterogeneity largely due to variable sialic acid content. The other variant appeared to be an asialo form with no affinity to concanavalin A. The two novel forms were of identical size and antigenicity compared to the normal enzyme. They might therefore reflect different post-translational modifications of the carbohydrate moiety of the enzyme.

Topics: Aged; Chromatography, Affinity; Colonic Neoplasms; Concanavalin A; Electrophoresis; Epitopes; Female; gamma-Glutamyltransferase; Genetic Variation; Humans; Male; Middle Aged; Rectal Neoplasms

The present study confirms previous observations that in vitro T lymphocyte proliferative response to mitogens is depressed in only some untreated patients with advanced or metastatic breast and colorectal cancer. Indomethacin, a prostaglandin synthetase inhibitor, at 1.0 microgram or 0.1 microgram/ml concentration significantly enhances the PHA, Con A or PWM response in these patients with breast and colorectal cancer (P less than 0.05 - P less than 0.01). Indomethacin has no mitogenic activity. Ethyl alcohol (0.01%), in which indomethacin is dissolved, also has no mitogenic or cytotoxic activity. Although the in vitro effect of indomethacin has been well-demonstrated, the in vivo effect of this agent on cell-mediated immunity in man has not yet been thoroughly investigated and thus, further studies of the effect of indomethacin administration on in vivo and in vitro cellular immunity seem warranted.

Topics: Adult; Aged; Breast Neoplasms; Colonic Neoplasms; Concanavalin A; Ethanol; Female; Humans; Indomethacin; Lymphocyte Activation; Male; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; Rectal Neoplasms; Stimulation, Chemical; T-Lymphocytes

In the present study we have evaluated monocyte phagocytosis and T lymphocyte in vitro reactivity to mitogens in peripheral blood samples from 14 colorectal cancer patients, from 21 nonneoplastic control patients, and from 22 normal donors. Monocyte and lymphocyte functions were tested before and 1 and 6 months after surgery. Our results indicate the existence of increased monocyte phagocytosis and decreased mitogen reactivity in untreated patients with advanced tumors. These abnormal responses persisted and were even more pronounced after surgery and chemotherapy.

Topics: Adenocarcinoma; Adult; Aged; Colonic Neoplasms; Concanavalin A; Female; Humans; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Monocytes; Phagocytosis; Phytohemagglutinins; Rectal Neoplasms; Rosette Formation

Blood-, lymph node-, and tumour-infiltrating lymphocytes (PBL, LNC, and TIL, respectively) from patients with colonic neoplasms were tested for responsiveness to phytohaemagglutinin (PHA). All populations responded, with LNC and PBL showing comparable reactivities while TIL were less reactive as assessed by incorporation of 3H-thymidine. Increased mitogen responsiveness was observed for T cells enriched by SRBC rosette formation or passage through nylon columns. Mitomycin C-treated LNC and TIL inhibited PHA induced 3H-thymidine incorporation of admixed autologous PBL, suggesting the presence of suppressor cells. Suppressor activity resided primarily in the SRBC rosetting population and was dose-dependent, with increasing numbers of LNC giving greater diminution of PHA response. Suppression by LNC was apparent only when they were added to PBL responders within 6 h of the initiation of stimulation assays, in common with the effects of Concanavalin A (Con A)-induced suppressors on PBL phytomitogen responsiveness. Con A-induced and LNC-suppressor activity could be reversed by addition of lymphocyte-conditioned medium (CM) containing T cell growth factor (TCGF; interleukin IL-2). These data provide further evidence that the suppressor phenomena observed in this system are a function of activated T cells present both in drainage lymph nodes and at the tumour site.

Topics: Cells, Cultured; Colonic Neoplasms; Concanavalin A; DNA Replication; Humans; Lymph Nodes; Lymphocyte Activation; Phytohemagglutinins; Rosette Formation; T-Lymphocytes; T-Lymphocytes, Regulatory

The histochemistry of mucin in normal large intestine and in experimental colorectal cancers induced in Wistar rats with 1,2-dimethylhydrazine was analyzed by modifications of the concanavalin A-horseradish peroxidase method (paradoxical concanavalin A-staining) and high-iron diamine-Alcian blue (pH 2.5) staining (HID-AB). Quantitative analysis of each mucin reaction was done by the use of an image analyzer. Regional variations in the percentage area of "labile class III mucin" were 1 to 50% in the distal colon and 50 to 90% in the proximal colon. Regional variations in the percentage area of sulfated sialomucin were 80 approximately 100% in the distal colon and under 20% in the proximal colon. The percentage area of "labile class III mucin" in primary colorectal cancers showed region-specific variation and that in metastatic colorectal cancers also showed similar region-specific variation of colorectal mucosa where primary tumors were located. In contrast, there were no region-specific mucin reactions for sulfated sialomucin in almost all primary and metastatic colorectal cancers.

Topics: 1,2-Dimethylhydrazine; Animals; Colon; Colonic Neoplasms; Concanavalin A; Dimethylhydrazines; Epithelium; Histocytochemistry; Intestinal Mucosa; Male; Mucins; Neoplasms, Experimental; Rats; Rats, Inbred Strains; Rectal Neoplasms

Lamina proprial lymphocytes (LPL), isolated by an EDTA-collagenase technique from patients with various colonic diseases, were investigated for LMIF release in vitro. On stimulation with the preparation of Kunin antigen, macrophage-depleted LPL from patients with severely or moderately active ulcerative colitis showed LMIF release which was significantly greater than that observed using LPL from patients with mild colitis or from those with other diseases of the large bowel, including Crohn's disease. Results similar to those obtained with LPL were found with the corresponding peripheral blood lymphocytes (PBL) stimulated by the preparation of Kunin antigen. In contrast, nonspecific stimulation in vitro with Concanavalin A showed no differences in LMIF releases by the LPL or PBL in the various disease groups. It is suggested that hypersensitivity to Kunin antigen may have pathogenic significance in ulcerative colitis.

Topics: Adolescent; Adult; Aged; Appendicitis; Colitis, Ulcerative; Colonic Diseases; Colonic Neoplasms; Concanavalin A; Crohn Disease; Edetic Acid; Female; Humans; Intestinal Mucosa; Leukocyte Migration-Inhibitory Factors; Lymphocytes; Lymphokines; Male; Microbial Collagenase; Middle Aged; Rectal Neoplasms; Rectal Prolapse

The response to mitogen (Con A) of the normal and cancer patient non-adherent cells (NAC) was studied. These cells were added to adherent cells (AC) monolayer in an autologous and a homologous combination and cultured with absence or presence of indomethacin. The mitogen response of patient autologous cells (NAC + AC) was poor and indomethacin did not cause any changes. The mitogen response of normal autologous cells (NAC + AC) was increased by indomethacin, and was dependent on the number of AC in the culture. NAC from patient and from control blood cultured alone did not show an increase of response to mitogen when indomethacin was in the culture. The patient NAC cultured with normal AC showed a low response and a slight increase of response in the presence of indomethacin. The suggestion that prostaglandins are not involved in the suppression of mitogen response of patient lymphocyte and that the patient lymphocytes are hyporesponsive to PGs is discussed.

Topics: Adult; Aged; Breast Neoplasms; Cell Adhesion; Colonic Neoplasms; Concanavalin A; Humans; Indomethacin; Lymphocyte Activation; Middle Aged; Neoplasms; Stomach Neoplasms

Topics: Carcinoembryonic Antigen; Colon; Colonic Neoplasms; Concanavalin A; Electrophoresis, Agar Gel; Glycoproteins; Hot Temperature; Humans; Immunodiffusion

Topics: Adenocarcinoma; Aged; Cell Adhesion; Colonic Neoplasms; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Male; Middle Aged; Phytohemagglutinins; Rectal Neoplasms; T-Lymphocytes

A methotrexate/concanavalin-A conjugate (MTX/Con-A) was prepared by covalent cross-linking. The extent of substitution was approximately 5 moles methotrexate (MTX) per mole concanavalin A (Con-A). Inhibition of dihydrofolate reductase by the conjugate was only marginal when compared to MTX. However, MTX/Con-A showed a 7- to 116 times higher cytocidal activity than MTX against cultured KB cells and various established cell lines from mouse colon adenocarcinomas 26 and 38, and Lewis lung carcinoma. The effect of MTX/Con-A was diminished when the conjugate was preincubated with alpha-methyl-D-mannoside, a specific binding sugar to Con-A. MTX/Con-A efficiently incorporated into cells and retained in them for longer periods of time than was MTX. The strong cytocidal action of the conjugate could be explained by the higher incorporation rate and the longer retention time of the conjugate in the cells.

Topics: Adenocarcinoma; Animals; Cell Line; Colonic Neoplasms; Concanavalin A; Methotrexate; Mice; Neoplasms, Experimental

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

A method is reported for the preparation of glutaraldehyde cross-linked horseradish peroxidase conjugates for the visualization of concanavalin A binding to frozen sections. The binding of concanavalin A to tissue sections of human colonic tumor is used to compare staining by horseradish peroxidase conjugates and monomers. Without decreasing specificity, the use of progressively larger molecular weight conjugates provides progressively greater intensity of staining of the epithelial components of colonic carcinomas and normal colons.

Topics: Aldehydes; Colonic Neoplasms; Concanavalin A; Glutaral; Horseradish Peroxidase; Humans; In Vitro Techniques; Molecular Weight; Protein Binding; Serum Albumin, Bovine; Staining and Labeling

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

Topics: Colon; Colonic Neoplasms; Concanavalin A; Epithelial Cells; Humans; In Vitro Techniques; Receptors, Drug

Active-specific immunotherapy with concanavalin A or with neuraminidase-modified syngeneic tumor cells has been studied in an experimental model of colon cancer. Systemic immunotherapy with concanavalin A-modified tumor cells or with neuraminidase-modified tumor cells has resulted in up to 70 percent cure of rats receiving a lethal inoculum of tumor in a model tumor system which otherwise proved 80 percent lethal to untreated hosts. The possible mechanisms whereby neuraminidase or concanavalin A are effective cell-surface modifiers for active-specific immunotherapy are discussed. In vitro studies suggest markedly heightened antigenic recognition following immunization with concanvalin A-modified syngeneic tumor cells. These studies represent the first apparent evidence for the definitive value of systemic active-specific immunotherapy for the adjuvant treatment of large bowel cancer.

Topics: Animals; Antigens, Neoplasm; Colonic Neoplasms; Concanavalin A; Disease Models, Animal; Immunization; Immunotherapy; Neoplasms, Experimental; Neuraminidase; Rats

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: 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 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

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

Topics: Acetamides; Adenocarcinoma; Agglutination; Animals; Carcinogens; Cell Differentiation; Cell Membrane; Colonic Neoplasms; Concanavalin A; Epithelium; Galactose; Glucosamine; Glucosyltransferases; Hydrazines; Intestinal Neoplasms; Intestines; Lectins; Neoplasms, Experimental; Rats; Rats, Inbred Strains; Transferases

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