concanavalin-a and Neoplasms

Concanavalin A (ConA), the most studied plant lectin, has been known as a potent anti-neoplastic agent for a long time. Since initial reports on its capacity to kill cancer cells, much attention has been devoted to unveiling the lectin's exact molecular mechanism. It has been revealed that ConA can bind to several receptors on cancerous and normal cells and modulate the related signaling cascades. The most studied host receptor for ConA is MT1-MMP, responsible for most of the lectin's modulations, ranging from activating immune cells to killing tumor cells. In this study, in addition to studying the effect of ConA on signaling and immune cell function, we will focus on the most up-to-date advancements that unraveled the molecular mechanisms by which ConA can induce autophagy and apoptosis in various cancer cell types, where it has been found that P73 and JAK/STAT3 are the leading players. Moreover, we further discuss the main signaling molecules causing liver injury as the most significant side effect of the lectin injection. Altogether, these findings may shed light on the complex signaling pathways controlling the diverse responses created via ConA treatment, thereby modulating these complex networks to create more potent lectin-based cancer therapy. Video Abstract.

Topics: Concanavalin A; Humans; Lectins; Matrix Metalloproteinase 14; Neoplasms; Plant Lectins

Plant lectin, a class of highly diverse non-immune origin and carbohydrate-binding proteins, has been reported to specially induce cancer cell through programmed cell death (PCD) pathways (apoptosis and/or autophagy), shedding lights on screening promising anti-cancer candidate agent for further therapeutic trials. However, the complicated molecular mechanisms by which plant lectins induced the programmed death of tumor cells, have not yet been fully clarified. Here, we summarized a novel model, based on vast amount of research, by which plant lectins eliminate various types of cancer cells via three major pathways, including a) direct ribosome inactivating, b) endocytosis-dependent mitochondrial dysfunction and c) sugar-containing receptors binding. A better understanding of the role of plant lectins played and further elucidation of the strategies targeting PCD would provide a new clue for the applications and modifications of plant lectin as a potential anti-cancer agent from bench to clinic.

Topics: Animals; Antineoplastic Agents; Apoptosis; Autophagy; Concanavalin A; HT29 Cells; Humans; Jurkat Cells; Models, Molecular; Neoplasms; Plant Lectins; Signal Transduction

Plant lectins, a group of highly diverse carbohydrate-binding proteins of non-immune origin, are ubiquitously distributed through a variety of plant species, and have recently drawn rising attention due to their remarkable ability to kill tumour cells using mechanisms implicated in autophagy. In this review, we provide a brief outline of structures of some representative plant lectins such as concanavalin A, Polygonatum cyrtonema lectin and mistletoe lectins. These can target autophagy by modulating BNIP-3, ROS-p38-p53, Ras-Raf and PI3KCI-Akt pathways, as well as Beclin-1, in many types of cancer cells. In addition, we further discuss how plant lectins are able to kill cancer cells by modulating autophagic death, for therapeutic purposes. Together, these findings provide a comprehensive perspective concerning plant lectins as promising new anti-tumour drugs, with respect to autophagic cell death in future cancer therapeutics.

Topics: Antineoplastic Agents; Autophagy; Concanavalin A; Humans; Neoplasms; Plant Lectins; Polygonatum; Signal Transduction; Viscum album

Concanavalin A (ConA), a Ca(2+)/Mn(2+)-dependent and mannose/glucose-binding legume lectin, has drawn a rising attention for its remarkable anti-proliferative and anti-tumor activities to a variety of cancer cells. ConA induces programmed cell death via mitochondria-mediated, P73-Foxo1a-Bim apoptosis and BNIP3-mediated mitochondrial autophagy. Through IKK-NF-κB-COX-2, SHP-2-MEK-1-ERK, and SHP-2-Ras-ERK anti-angiogenic pathways, ConA would inhibit cancer cell survival. In addition, ConA stimulates cell immunity and generates an immune memory, resisting to the same genotypic tumor. These biological findings shed light on new perspectives of ConA as a potential anti-neoplastic agent targeting apoptosis, autophagy and anti-angiogenesis in pre-clinical or clinical trials for cancer therapeutics.

Topics: Amino Acid Sequence; Angiogenesis Inhibitors; Antineoplastic Agents; Apoptosis; Autophagy; Cell Survival; Concanavalin A; Humans; MAP Kinase Signaling System; Molecular Sequence Data; Neoplasms; Protein Tyrosine Phosphatase, Non-Receptor Type 11; ras Proteins

Plant lectins, carbohydrate-binding proteins distributed widely in a variety of plant species, have been well-known to possess a broad range of significant biological functions such as anti-tumor, anti-fungal and anti-viral activities. Amongst the seven major lectin families, legume lectins have been drawing a rising attention for cancer biologists due to their remarkable anti-tumor properties compared to other lectin families. In this review, we mainly focus on analyzing the anti-tumor activities of Concanavalin A (ConA), the first and most typical representative of legume lectin family, and its related mechanisms of cell death implicated in apoptosis and autophagy. We present the up-to-date experimental advancements that ConA is able to induce cancer cell apoptosis through mitochondria-dependent and p73-mediated pathways, as well as ConA can induce cancer cell autophagy through a mitochondria-dependent signaling pathway. In addition, we further discuss the pre-clinical studies of ConA for its potential cancer therapeutic applications. In conclusion, these findings may shed light on the complicated molecular mechanisms of ConA-induced cancer cell death, thereby opening a new perspective for plant lectins as potential anti-neoplastic drugs in future cancer therapeutics.

Topics: Antineoplastic Agents; Apoptosis; Autophagy; Concanavalin A; DNA-Binding Proteins; Humans; Mitochondria; Neoplasms; Nuclear Proteins; Plant Lectins; Tumor Protein p73; Tumor Suppressor Proteins

Topics: Concanavalin A; HIV Infections; Humans; Immunization; Mitogens; Neoplasms; Phytohemagglutinins; Pokeweed Mitogens; Skin Tests

Interleukin-2 (IL-2) is a type of the lymphokine which Morgan et al. found in 1976 to be a specific growth factor of T lymphocytes. Initially, it was called the T cell growth factor (TCGF), but it was renamed interleukin-2 (IL-2) in 1979. IL-2 is an essential factor for the differentiation and growth of T cell and NK cell, and it is involved in the adaptive immune reaction through such cells. Thus, IL-2 can be expected to play an important role in the improvement of the immunologic adaptive function in various immune system failure type disease and in malignant tumors among others. With the recent production of recombinant IL-2, the latter's biological and immunological functions are gradually coming to light, and its clinical applicability is also being seen in terms of effectiveness. Thus, administration of IL-2 alone, the induction of IL-2-dependent, tumor-specific CTL (cytotoxic T lymphocyte), or the induction and growth by IL-2 of lymphokine-activated killer (LAK) cell, have paved the way for adoptive immunotherapy. The clinical phase I study of IL-2 (TGP-3) has been completed, and the second phase study is now in progress. A pilot study is also under way using LAK cells.

Topics: Animals; Antibody-Dependent Cell Cytotoxicity; Concanavalin A; Humans; Infusions, Parenteral; Interleukin-2; Killer Cells, Natural; Lymphocyte Activation; Neoplasms; T-Lymphocytes

Tumor promoters act on carcinogen-initiated tissues to cause phenotypic expression of malignancy. Phorbol ester tumor promoters, like hormones, act on cells and tissues at nanomolar concentrations, often producing "physiological" effects. These promoters also act on cells to produce what appear to be nonphysiologic or toxic effects. One of the major questions regarding these phenomena is, Which action or actions of promoters are important in phenotypic expression of malignancy?

Topics: Animals; Calcimycin; Calcium; Chromosomes; Concanavalin A; Cyclic GMP; DNA; Egtazic Acid; Humans; Myristates; Neoplasms; Potassium; Protein Kinase C; Protons; Sodium; Tetradecanoylphorbol Acetate

Topics: Cell Movement; Chemotactic Factors; Chemotaxis, Leukocyte; Concanavalin A; Dinoprostone; Humans; In Vitro Techniques; Lymphocytes; Neoplasms; Prostaglandins E; T-Lymphocytes; T-Lymphocytes, Regulatory

On exogenous stimulus immunocompetent leucocytes produce antigen-specific factors, which essentially determine the magnitude and duration of T-lymphocyte dependent immunoreactions. Interleukin 1 (IL-1, monokin) and interleukin 2 (IL-2, lymphokine) form a bimodal amplification system which may operate in vivo at the level of peripheral lymphoid organs, which interleukin 3 (IL 3, lymphokine) may function as a positive feed-back signal at the level of multipotential stem cells. The physiological IL-2 has wider importance, since the permanent expression of Il-2 gene due to the insertion of the viral promoter sequence (HTLV in human) led to uncontrolled proliferation of T-cells and to the development of lymphomas and leukemias of mature T-cell phenotype. On the afferent arc of immunoreactions endogenous factors mediate T-cell proliferation inhibitory effect probably via interaction with the interleukin system. Some practical aspects of these two antagonistic group of factors are presented and discussed.

Topics: Animals; Cell Line; Concanavalin A; Deltaretrovirus; DNA, Recombinant; Humans; Immune Tolerance; Interleukin-1; Interleukin-2; Interleukin-3; Leukemia; Lymphocyte Activation; Lymphokines; Lymphoma; Macrophages; Mice; Neoplasms; Phytohemagglutinins; T-Lymphocytes

Topics: Animals; Blood Platelets; Carbohydrate Metabolism; Concanavalin A; Erythrocytes; Fucose; Galactosamine; Galactose; Glucosamine; Glycoproteins; Humans; Lectins; Lymphocytes; Mannose; Membrane Proteins; Neoplasms; Neurons; Protein Binding

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

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

Topics: Agglutination; Animals; Antibodies, Neoplasm; Antibody Formation; Antigen-Antibody Complex; Antigens, Neoplasm; Cell Membrane; Concanavalin A; Ferritins; Fluorescent Antibody Technique; Immunoglobulin G; Ligands; Lymphocytes; Macromolecular Substances; Mice; Microscopy, Electron; Models, Biological; Neoplasms; Phosphatidylcholines; Pinocytosis; Receptors, Drug

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

Autoimmune hepatitis (AIH), characterized by immune-driven liver destruction and cytokine production, is a progressive inflammatory liver condition that may progress to hepatic cirrhosis or tumors. However, the underlying mechanism is not well understood, and the treatment options for this disease are limited. Pemetrexed (PEM), a clinically used anti-folate drug for treating various tumors, was found to inhibit the nuclear factor (NF)-κB signaling pathways that exert an important role in the development of AIH. Here, we investigated the impact of PEM on immune-mediated hepatic injuries using a murine model of Concanavalin A (Con A)-induced hepatitis, a well-established model for AIH. Mice received intraperitoneal PEM injections 3 times at 12-hour intervals, and two hours later, they were challenged with Con A. Liver samples and serum were collected after 10 h. The results indicate that PEM significantly improved mouse survival rates and lowered serum transaminase levels. Moreover, PEM effectively alleviated oxidative stress, reduced histopathological liver damage, and mitigated hepatocyte apoptosis. Notably, it reduced the activation of M1-type macrophages in the liver. The expression of proinflammatory cytokines and genes associated with M1 macrophages, such as tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-12, IL-1β, and inducible nitric oxide synthase (iNOS), was also decreased. Finally, the results indicated that PEM regulates M1 macrophage activation by modulating the NF-κB signaling pathways. Overall, these results demonstrate that PEM effectively guards against immune-mediated hepatic injuries induced by Con A by inhibiting M1 macrophage activation through the NF-κB signaling pathways and indicate the potential of PEM as a practical treatment option for AIH in clinical settings.

Topics: Animals; Concanavalin A; Cytokines; Hepatitis, Autoimmune; Interleukin-6; Macrophage Activation; Mice; Neoplasms; NF-kappa B; Pemetrexed

Topics: Cell Line, Tumor; Concanavalin A; Drug Carriers; Drug Delivery Systems; Drug Liberation; Gelatin; Nanoparticles; Neoplasms

Receptor tyrosine kinases (RTK) have been the most prevalent therapeutic targets in anti-cancer drug development. However, the emergence of drug resistance toward single target RTK inhibitors remains a major challenge to achieve long-term remissions. Development of alternative RTK inhibitory strategies that bypass drug resistance is much wanted. In the present study, we found that selected cell surface RTKs were inhibited and crosslinked into detergent resistant complexes by oligomeric but not monomeric concanavalin A (ConA). The inhibition of RTKs by ConA led to suppression of pro-survival pathways and induction of apoptosis in multiple cancer cell lines, while overexpression of constitutively activated protein kinase B (AKT) reversed the apoptotic effect. However, major cell stress sensing checkpoints were not influenced by ConA. To our knowledge, selective crosslinking and inhibition of cell surface receptors by ConA-like molecules might represent a previously unidentified mechanism that could be potentially exploited for therapeutic development.

Topics: Apoptosis; Cell Line, Tumor; Concanavalin A; Drug Resistance, Neoplasm; Humans; Neoplasms; NF-kappa B; Protein Conformation; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Receptor Protein-Tyrosine Kinases; Signal Transduction

In this study, we investigated the chemical and biological profile of lectin isolated from Japanese red sword beans (Canavalia gladiata; RSBs). RSB lectin was purified using maltamyl-Sepharose 4B and subjected to amino acid composition and partial amino acid sequencing analyses, and evaluated for blood and carbohydrate specificity, mitogenic activity, splenic natural killer (NK) cell activity, and its effect on B16 melanoma cell proliferation, compared with Concanavalin A (Con A). The amino acid composition and sequences of RSB lectin were similar to those of Con A. RSB lectin showed specificity to mannose, glucose, maltose, methyl-D-mannoside, and thyroglobulin, but not rhamnose, using mouse, sheep, and rabbit erythrocytes. Compared with Con A, RSB lectin showed low resistance to proteases and to temperatures greater than 70 °C, but high mitogenic activity for mouse splenic cells. Notably, while treatment with RSB lectin and Con A (0.01 and 0.1 μg/mL) promoted similar levels of splenic NK cell activity, which were higher than that observed in the control (0 μg/mL) and interleukin 2 (IL-2) (25 U)-treated populations, RBS lectin exerted a significantly stronger anti-proliferative effect than Con A at a concentration of 125.0 μg per well. Overall, our results show that RSB lectin might exert immunological effects on mouse splenic cells and could thus be used as a potential cancer chemopreventive agent.. Japanese red sword bean (RSB) is a tropical perennial legume consumed in many Asian countries. RSB lectin shows specificity to mannose, glucose, maltose, methyl-d-mannoside, and thyroglobulin, but not to rhamnose, using mouse, sheep, and rabbit erythrocytes. RSB lectin exhibits similarities to Concanavalin A in amino acid composition and sequence, shows mitogenic activity for mouse splenic cells and strong anti-proliferative activity for B16 melanoma cells, and also enhances the activity of splenic natural killer (NK) cells against YAC-1 cells. Thus, RSB lectin has the potential to be used as a bioactive protein in medical research.

Topics: Animals; Canavalia; Cell Line, Tumor; Cell Proliferation; Chemoprevention; Concanavalin A; Erythrocytes; Fabaceae; Glucose; Killer Cells, Natural; Lectins; Maltose; Mannose; Methylmannosides; Mice; Neoplasms; Plant Extracts; Rabbits; Rhamnose; Sheep; Thyroglobulin

Multifunctional nanocomposites containing intrinsic property for serving as the sensing elements as well as targeted nanoconjugates are highly preferred in various therapeutic applications. In this work, nanocomposites of graphene quantum dots (GQDs) and Fe

Topics: Cell Survival; Concanavalin A; Doxorubicin; Drug Delivery Systems; Electrodes; Endothelial Cells; Ferrosoferric Oxide; Graphite; HeLa Cells; Humans; Limit of Detection; Nanocomposites; Neoplasms; Platinum; Quantum Dots

The successful development of selective detection of cancer cells from normal cells is a highly demanded but challenging task. Herein, we have developed a rapid and label-free impedimetric biosensor for quantitative determination of cancer cells with high glycoprotein expression. Homogenously distributed 2-9 nm graphene quantum dots (GQDs) was anchored on the surface Fe

Topics: Biosensing Techniques; Carbohydrates; Cell Line; Concanavalin A; Electric Impedance; Electrodes; Glycoproteins; Graphite; HeLa Cells; Humans; Immobilized Proteins; MCF-7 Cells; Neoplasms; Quantum Dots

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

Tumor-marker-imprinted hydrogels having lectin and antibody molecules as ligands for a tumor-specific marker glycoprotein were strategically prepared by biomolecular imprinting using minute amounts of low-molecular-weight or high-molecular-weight cross-linkers. The tumor-marker-imprinted hydrogels shrank gradually in response to a target glycoprotein, because their apparent cross-linking density increased owing to simultaneous complex formation of lectin and antibody ligands with a target glycoprotein after their ligands dynamically recognized the glycoprotein. The swelling ratio of the tumor-marker-imprinted hydrogel using high-molecular-weight cross-linker with an optimal chain length decreased more drastically than that using a low-molecular-weight cross-linker, but the hydrogel using the cross-linker with a chain that was too long did not exhibit tumor-marker responsive behavior. This paper focuses on the effect of the molecular weight of cross-linkers on the responsive behavior of tumor-marker-imprinted hydrogels having lectin and antibody molecules as ligands. The cross-linker chain length was an important factor in determining the dynamic glycoprotein recognition and responsive behavior of the biomolecule-imprinted hydrogels.

Topics: Acrylamides; Adsorption; alpha-Fetoproteins; Antibodies, Neoplasm; Biomarkers, Tumor; Compressive Strength; Concanavalin A; Cross-Linking Reagents; Humans; Hydrogels; Methacrylates; Models, Molecular; Molecular Imprinting; Molecular Weight; Neoplasms; Polyethylene Glycols

In this article, we report a novel lectin-based biosensor for electrochemical assay of cancer-associated glycosylation by comparative study of mannose and sialic acid expression on normal and cancer cells derived from human lung, liver, and prostate. Using a sandwich format, high sensitivity and selectivity were achieved by combining the lectin-based biosensor with the {lectin-Au-Th} bioconjugates featuring lectin and thionine (Th) labels linked to gold nanoparticles (AuNPs) for signal amplification. The proposed strategy demonstrated that mannose exhibited high expression levels in both normal and cancer cells, while sialic acid was more abundant in cancer cells as compared to normal ones. The results were in good agreement with those from fluorescent microscopy studies. The differences in the two glycan expression indicated that sialic acid could serve as a potential biomarker for early cancer detection. The lectin-based biosensor was also successfully used to quantify cancer cells and evaluate the average amount of sialic acid on single cell surface, which could supply significant information on glycan functions in cancer progression. Overall, the lectin-based electrochemical biosensor provides an effective pathway to analyze glycan expression on living cells and may greatly facilitate the medical diagnosis and treatment in early process of cancer.

Topics: Biosensing Techniques; Cell Line, Tumor; Cell Survival; Concanavalin A; Electrochemistry; Electrodes; Glass; Glycosylation; Gold; Humans; Immobilized Proteins; Mannose; Metal Nanoparticles; Microscopy, Fluorescence; N-Acetylneuraminic Acid; Nanotubes, Carbon; Neoplasms; Phenothiazines; Plant Lectins; Protein Stability; Ribosome Inactivating Proteins

Concanavalin A (Con A), a mannose/glucose-binding legume lectin, can induce cancer cell death through a mitochondria-mediated autophagic pathway; however, the precise mechanisms by which it mediates cell death are still only rudimentarily understood. In the present study, Con A possesses a remarkable antiproliferative effect on human melanoma A375 cells. Also, there is a link between the antiproliferative activity of Con A and its sugar-binding activity. Subsequently, Con A can induce human melanoma A375 cell apoptosis in a caspase-dependent manner. In addition, the treatment with Con A can cause mitochondrial transmembrane potential collapse, leading to cytochrome c release and caspase-9-caspase-3 activation. In conclusion, we demonstrate that there may be a close correlation between the antiproliferative activity of Con A and its sugar-binding activity. More importantly, we report for the first time that Con A can induce human melanoma A375 cell death in a caspase-dependent manner as well as via a mitochondrial apoptotic pathway.

Topics: Apoptosis; Carbohydrates; Caspases; Cell Line, Tumor; Cell Proliferation; Concanavalin A; Cytochromes c; Enzyme Activation; Humans; Mitochondria; Mitogens; Neoplasms

The majority of patients with advanced cancer are recognised by impaired immune competence influenced by several factors, including the type and stage of the tumour and the presence of cachexia. Recently, a specific nutritional combination containing fish oil, specific oligosaccharide mixture, high protein content and leucine has been developed aimed to support the immune system of cancer patients in order to reduce the frequency and severity of (infectious) complications. In a recently modified animal model cachexia is induced by inoculation of C26 tumour cells in mice. In a pre-cachectic state, no effect was observed on contact hypersensitivity, a validated in vivo method to measure Th1-mediated immune function, after adding the individual nutritional ingredients to the diet of tumour-bearing mice. However, the complete mixture resulted in significantly improved Th1 immunity. Moreover, in a cachectic state, the complete mixture reduced plasma levels of pro-inflammatory cytokines and beneficially affected ex vivo immune function. Accordingly, the combination of the nutritional ingredients is required to obtain a synergistic effect, leading to a reduced inflammatory state and improved immune competence. From this, it can be concluded that the specific nutritional combination has potential as immune-supporting nutritional intervention to reduce the risk of (infectious) complications in cancer patients.

Topics: Animal Feed; Animals; Cachexia; Cell Line, Tumor; Cell Proliferation; Concanavalin A; Cytokines; Dinoprostone; Disease Models, Animal; Fish Oils; Lipopolysaccharides; Male; Mice; Neoplasms; Oligosaccharides; Spleen; T-Lymphocytes

Combinations of cellular immune-based therapies with chemotherapy and other antitumour agents may be of significant clinical benefit in the treatment of many forms of cancer. Gamma delta (gammadelta) T cells are of particular interest for use in such combined therapies due to their potent antitumour cytotoxicity and relative ease of generation in vitro. Here, we demonstrate high levels of cytotoxicity against solid tumour-derived cell lines with combination treatment utilizing Vgamma9Vdelta2 T cells, chemotherapeutic agents and the bisphosphonate, zoledronate. Pre-treatment with low concentrations of chemotherapeutic agents or zoledronate sensitized tumour cells to rapid killing by Vgamma9Vdelta2 T cells with levels of cytotoxicity approaching 90%. In addition, zoledronate enhanced the chemotherapy-induced sensitization of tumour cells to Vgamma9Vdelta2 T cell cytotoxicity resulting in almost 100% lysis of tumour targets in some cases. Vgamma9Vdelta2 T cell cytotoxicity was mediated by perforin following TCR-dependent and isoprenoid-mediated recognition of tumour cells. Production of IFN-gamma by Vgamma9Vdelta2 T cells was also induced after exposure to sensitized targets. We conclude that administration of Vgamma9Vdelta2 T cells at suitable intervals after chemotherapy and zoledronate may substantially increase antitumour activities in a range of malignancies.

Topics: Adenocarcinoma; Antineoplastic Agents; Apoptosis; Breast Neoplasms; Burkitt Lymphoma; Carcinoma; Cell Line, Tumor; Cisplatin; Colorectal Neoplasms; Concanavalin A; Cytotoxicity, Immunologic; Diphosphonates; Doxorubicin; Drug Screening Assays, Antitumor; Drug Synergism; Etoposide; Female; Genes, T-Cell Receptor delta; Genes, T-Cell Receptor gamma; Humans; Imidazoles; Interferon-gamma; Lovastatin; Lung Neoplasms; Male; Membrane Glycoproteins; Neoplasms; Perforin; Pore Forming Cytotoxic Proteins; Prostatic Neoplasms; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocyte Subsets; Urinary Bladder Neoplasms; Vincristine; Zoledronic Acid

In this study, we investigated the effect of Rauwolfia radix on heat shock protein (HSP) 70 expression and cytotoxicity against tumor cells in activated human T cells. When activated T cells were cultured with Rauwolfia radix for 18 h, HSP70 expression after heat shock was remarkably increased, and cytotoxicity against T98G tumor cells was augmented. Moreover, Rauwolfia radix also enhanced the cytotoxicity of heat shocked activated T cells against Molt-4 and T98G tumor cells. Secretions of interferon-gamma (IFN-gamma) and tumor necrosis alpha (TNF-alpha), due to Concanavalin A (Con A) stimulation, were increased by Rauwolfia radix in activated T cells. To investigate the antitumor effect in vivo, EL-4 tumor-bearing mice were administered with Rauwolfia radix in drinking water. The survival period of the Rauwolfia radix treatment group was significantly prolonged compared with that of the control group. Reserpine, the major active ingredient of Rauwolfia radix, also enhanced the cytotoxicity of activated T cells against Molt-4 and T98G tumor cells, and prolonged the survival period of EL-4 tumor-bearing mice. Taken together, our results suggest that Rauwolfia radix can enhance the activity of immune cells against tumor cells.

Topics: Animals; Blotting, Western; Cell Survival; Concanavalin A; Cytotoxicity, Immunologic; Female; Gene Expression Regulation; Heat-Shock Response; HSP70 Heat-Shock Proteins; Humans; Interferon-gamma; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Neoplasms; Plant Extracts; Plant Roots; Rauwolfia; Reserpine; T-Lymphocytes; Time Factors; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

The specificity of monoclonal antibodies against gastric mucins (designated as HIK1083, PGM 36, and PGM 37) was studied immunohistochemically in normal, metaplastic, and neoplastic human tissues. These antibodies labeled class III mucin-producing cells identified by paradoxical concanavalin A staining in normal stomach, duodenum (Brunner gland), biliary tract, and main pancreatic duct; in mucinous metaplasia of pancreas and gallbladder; and in adenocarcinomas of stomach (90%), bile duct (80%), gallbladder (100%), pancreas (80%), lung (100% of goblet cell type adenocarcinomas), ovary (67% of mucinous carcinomas), and uterine cervix (100% of adenoma malignum tumors). Normal and neoplastic cells of esophagus, colon, salivary gland, kidney, endometrium, breast, prostate, and liver, as well as normal small intestine, lung, and uterine cervix, were all negative. The antibodies used should be valuable for the detection of class III mucin and class III mucin-producing cells in normal, metaplastic, and neoplastic tissues.

Topics: Antibodies, Monoclonal; Cell Line; Concanavalin A; Female; Gastric Mucosa; Humans; Male; Metaplasia; Mucins; Neoplasms; Reference Values; Staining and Labeling

The anti-tumor activity of human peripheral blood mononuclear cells (PBMC) against various tumor cell line cells (K562, Daudi, KMG-2, and KATOIII) was enhanced by coculture with irradiated BALL-1, but not with other irradiated B cell line cells (NALM-1, Namalwa, and Daudi). PBMC cocultured with BALL-1, however, failed to exhibit evident cytotoxicity against autologous concanavalin A-induced lymphoblasts. The enhancement of the anti-tumor activity seemed not to be correlated with EBNA and HLA-DR expression on B cell line cells. Monoclonal antibodies (mAbs) against interleukin (IL)-2, interferon-gamma, IL-12, IL-15, tumor necrosis factor-alpha and lymphotoxin showed little or no suppression of the anti-tumor activity of PBMC treated with irradiated BALL-1. Furthermore, the culture supernatants of BALL-1 failed to enhance the anti-tumor activity of PBMC, suggesting no involvement of soluble factors in the induction of the anti-tumor activity. The anti-tumor activity of PBMC treated with BALL-1 was synergistically enhanced by an additional IL-2 stimulation. Periodate-lysine-paraformaldehyde-fixed, but not ethanol- or acetone-fixed, BALL-1 could significantly enhance the anti-tumor activity. Furthermore, BALL-1-derived membrane fraction, but not that of Daudi, enhances the anti-tumor activity. It was thus suggested that some membrane glycoproteins on the cell surface of BALL-1 play a crucial role in the induction of the anti-tumor activity. By analysis using mAbs against human leukocytes, we found that depletion of CD11b, CD16, and CD56-positive cells resulted in decreased anti-tumor activity, suggesting that the main effector cells in the BALL-1-induced anti-tumor activity were natural killer (NK) cells. The present results thus raise the possibility that BALL-1, probably via membrane glycoproteins, modulates NK cell-mediated anti-tumor activity.

Topics: Antibodies, Monoclonal; Coculture Techniques; Concanavalin A; Cytotoxicity, Immunologic; Humans; Immunologic Factors; Interferon-gamma; Interleukin-2; Killer Cells, Natural; Lymphocyte Subsets; Lymphocytes; Lymphoma, B-Cell; Neoplasms; Tissue Fixation; Tumor Cells, Cultured

ICR mice were treated with a carcinogen, N-butyl-N'-butanolnitrosoamine BBN), every day for 8 consecutive weeks and the effects of oral administration of edible mushrooms on the induction of urinary bladder carcinoma and on the activities of macrophages and lymphocytes were studied. Bladder carcinoma were found in all 10 mice (100%) treated with BBN alone, while we observed carcinoma only in 9 of 17 mice (52.9%), in 7 of 15 mice (46.7%) and 13 of 20 mice (65.0%) treated with Lentinus edodes, Grifola frondosa and Pleurotus ostreatus, respectively. Chemotactic activity of macrophages was suppressed in mice treated with BBN alone but maintained almost the normal level in mice treated with BBN plus Lentinus, Grifola or Pleurotus. Lymphocytes collected from mice treated with BBN plus each mushroom showed almost normal blastogenic response against concanavalin A, although those from mice treated with BBN alone completely retarded their response. Cytotoxic activity of lymphocytes against Yac-1 cells was also maintained at a normal level in mice treated with BBN plus each mushroom. Whereas in mice treated with BBN alone significant depression of NK cell activity occurred. Significantly higher cytotoxic activity against P-815 cells was observed in lymphocytes from mice treated with BBN plus each mushroom than that in lymphocytes from normal mice or mice treated with BBN alone.

Topics: Animals; Antibiotics, Antineoplastic; Basidiomycota; beta-Glucans; Butylhydroxybutylnitrosamine; Carcinogens; Cells, Cultured; Chemotaxis; Concanavalin A; Cytotoxicity, Immunologic; Disease Outbreaks; Eating; Female; Glucans; Lentinan; Lymphocyte Activation; Lymphocytes; Macrophages; Medicine, Traditional; Mice; Mice, Inbred ICR; Neoplasms; Polyporaceae; Polysaccharides; Spleen; Urinary Bladder Neoplasms

Molecular size is one of the key determinants of transvascular transport of therapeutic agents in tumors. However, there are no data in the literature on the molecular size dependence of microvascular permeability in tumors. Therefore, we measured microvascular permeability to various macromolecules in the human colon adenocarcinoma LS174T transplanted in dorsal skin chambers in severe combined immunodeficient mice. These molecules were fluorescently labeled and injected i.v. into mice. The microvascular permeability was calculated from the fluorescence intensity measured by the intravital fluorescence microscopy technique. The value of permeability varied approximately 2-fold in the range of molecular weight from 25,000 to 160,000. These data indicate that tumor vessels are less permselective than normal vessels, presumably due to large pores in the vessel wall. The transport of macromolecules appears to be limited by diffusion through these pores. The cutoff size of the pores was estimated by observations of transvascular transport of sterically stabilized liposomes of 100-600 nm in diameter. We found that tumor vessels in our model were permeable to liposomes of up to 400 nm in diameter, suggesting that the cutoff size of the pores is between 400 and 600 nm in diameter.

Topics: Animals; Capillary Permeability; Concanavalin A; Diffusion; Humans; Immunoglobulin Fab Fragments; Immunoglobulin Fc Fragments; Immunoglobulin G; Liposomes; Mice; Mice, SCID; Molecular Weight; Neoplasms; Ovalbumin; Transplantation, Heterologous

Lipogenesis is essential for the rapid proliferation of cells and it is established that the biosynthesis of selected lipids precedes S-phase, DNA synthesis and the initiation of cell division. Pyruvate was previously shown to be an important lipid precursor for lymphocytes, macrophages and tumour cells. This study now reports the role of prostaglandins (PG) on the regulation of lipogenesis from [3-14C] pyruvate in 24-h cultured lymphocytes. It is shown that indomethacin (10 microM) and ibuprofen (10 microM), both inhibitors of PG biosynthesis, increased [3-14C] pyruvate incorporations into phospholipid and cholesterol fractions in resting lymphocytes but reduced its incorporation into these fractions in concanavalin A (Con A)-stimulated lymphocytes and tumour cells. These two agents also affected [2-14C] thymidine incorporation into the DNA of these cells in the same manner. Lipids produced from [3-14C] pyruvate and exported into the cell culture medium were also measured. The PG biosynthesis inhibitions reduced the transfer to culture medium of arachidonic acid, phospholipids, cholesterol and fatty acids higher than C20 by lymphocytes and tumour cells. Although macrophages are not proliferative cells, the cytoplasmic export measurement is important because these cells have a high capacity for lipid secretion. The results show that the PG biosynthesis inhibitors do not affect the export of phospholipids and cholesterol in macrophages. They do however markedly change the export of arachidonic acid and fatty acids higher than C20 produced from [3-14C] pyruvate in macrophages. It is also reported that a cell-stimulating response diminished the above fatty acid outcome in resting cells and augmented it in thioglycollate-stimulated macrophages. The findings suggest that PG modulation of lipogenesis may depend on cell cycle phase as well as the intrinsic lipid metabolic diversity and capacity.

Topics: Acetates; Animals; Arachidonic Acid; Carbon Radioisotopes; Cells, Cultured; Concanavalin A; DNA; DNA, Neoplasm; HeLa Cells; Humans; Ibuprofen; Indomethacin; KB Cells; Lipids; Lymphocyte Activation; Lymphocytes; Macrophages, Peritoneal; Neoplasms; Neoplasms, Experimental; Prostaglandins; Pyruvates; Pyruvic Acid; Rats; Thymidine; Tumor Cells, Cultured

We developed an assay for the direct determination of selenium in serum with a Perkin-Elmer Model 4100ZL Zeeman atomic absorption spectrometer and Ag-Cu-Mg modifier. We used this assay to analyze concanavalin A-bound selenoprotein (CABSP) in human serum after concanavalin A (ConA) affinity chromatography. The CABSP was identified as a single-chain glycoprotein of 57.3 kDa. Carbohydrate units were N- and O-linked to the protein. The selenium moiety was selenocysteine. Total selenium, glutathione peroxidase (GPX; EC 1.11.1.9), ConA-bound selenium (CABS), and alpha 1-acid glycoprotein (AAG) were determined in normal subjects and patients with various pathological conditions. CABS accounted for 44.1% +/- 6.3% of total selenium in sera from normal subjects and 46.5% +/- 3.9% to 55.1% +/- 8.1% in sera from patients with a variety of diseases. Total selenium in serum was well correlated with serum CABS (r = 0.860), but not with serum GPX activity (r = 0.117), for all patients studied. Serum CABS increased in normal subjects after selenium supplementation. Serum CABSP did not behave as an acute-phase reactant, compared with AAG.

Topics: Adult; Arthritis, Rheumatoid; Chromatography, Affinity; Chromatography, Gel; Concanavalin A; Diabetes Mellitus; Glutathione Peroxidase; Glycosylation; Humans; Kidney Diseases; Middle Aged; Neoplasms; Orosomucoid; Proteins; Reference Values; Selenium; Selenoproteins; Spectrophotometry, Atomic

We studied in a homologous system the procoagulant activity of human tumor cells cultured "in vitro" (1402 primary melanoma, Me 7110/2 metastatic melanoma, Hep G2 hepatoma and GLC1 small cell lung carcinoma) or of cells freshly isolated from different human tumor tissues. Tumor cells cultured "in vitro" possessed and released a factor VII dependent procoagulant activity, which was inhibited by concanavalin A and unaffected by iodoacetamide or HgCl2. The activity released by the cells of metastatic melanoma was higher than that released by the cells of the primary tumor. On the contrary, cancer cells isolated from tumor tissues possessed and released a factor VII independent activity which was inhibited by iodoacetamide of HgCl2 and was not modified by concanavalin A. Therefore, different methods for the preparation of tumor cell suspensions have to be used for the study of tumor procoagulants, since their expression depends very largely on the source of tumor cells. Furthermore, cultured human tumor cells are not an appropriate model for the "in vivo" procoagulant effect of tumor cells.

Topics: Blood Coagulation Factors; Concanavalin A; Cysteine Endopeptidases; Factor VII; Factor X; Humans; Iodoacetamide; Melanoma; Mercuric Chloride; Neoplasm Proteins; Neoplasms; Neoplastic Stem Cells; Tumor Cells, Cultured

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

Tumor necrosis factor (TNF), a protein predominantly produced by activated macrophages/monocytes, is presently available in recombinant, purified form for clinical trials. Intensive studies in many laboratories have shown that besides the tumorcytotoxic effects, TNF acts on a large array of different cells and has potent immunomodulatory activities. In a clinical phase I study, some immunologic functional parameters of blood cells from patients who received 24-hour infusions of recombinant human TNF (rhTNF) were analyzed. Natural killer (NK) cell activity, TNF production, interleukin-1 (IL-1) production and mitogen-induced proliferation were measured either in whole blood samples or in cultures of peripheral mononuclear leukocytes of the patients directly before and after rhTNF infusion. NK cell activity, TNF and IL-1 production capacity and proliferative responses to concanavalin A (Con A) were significantly reduced after rhTNF application. We conclude from these observations that rhTNF in vivo acts directly or indirectly on NK cells and monocytes by either inactivating their functional capacity or by absorbing the relevant cells to the endothelial cell layer, thus removing them from circulation.

Topics: Adult; Concanavalin A; Cytotoxicity, Immunologic; Drug Evaluation; Female; Humans; Immunosuppressive Agents; Infusions, Intravenous; Interleukin-1; Killer Cells, Natural; Lymphocyte Activation; Male; Middle Aged; Neoplasms; Phytohemagglutinins; Receptors, Fc; Receptors, IgG; Recombinant Proteins; Tumor Necrosis Factor-alpha

Mediated agglutination of erythrocytes was studied in the presence of concanavalin A (Con A). Agglutinability of erythrocytes from cancer patients was found to be higher than that of normal cells as a result of chemical factors present in the blood plasma which transform the cell surface, the number of Con A receptors remaining unchanged. It is concluded that increased agglutinability of erythrocytes from cancer patients is mainly due to a decrease in the negative charge of cells and the appearance of echinocytic forms.

Topics: Agglutination; Concanavalin A; Erythrocytes; Humans; Neoplasms

Low dose cyclophosphamide (CY) can augment the development of delayed-type hypersensitivity to primary antigens in patients with advanced cancer. In this paper, we have considered the hypothesis that the immunopotentiation is related to reduction of T-suppressor activity. Peripheral blood lymphocytes were collected and cryopreserved from 45 patients with metastatic malignancy before and then 3, 7, and 19 days after administration of CY, 300 mg/m2 i.v. The peripheral blood lymphocytes were tested for generation of concanavalin A-inducible suppressor activity, proliferative response to phytohemagglutinin, and phenotype using monoclonal antibodies to CD4 and CD8. Concanavalin A-inducible suppression was significantly reduced by day 3 and declined progressively through day 19. The mean percentage changes in suppression were: day 3, -23.4 +/- 6.8 (SE) (P less than 0.01); day 7, -33.1 +/- 14.3 (P = 0.052); day 19, -43.1 +/- 10.7 (P less than 0.01). In contrast, CY caused no significant changes in phytohemagglutinin proliferation (mean percentage changes: day 3, -4.7 +/- 6.1; day 7, -15.6 +/- 7.5; day 19, -5.5 +/- 8.1), indicating that the reduction in concanavalin A-inducible suppression was not merely a reflection of a general reduction in peripheral blood lymphocyte function. The total number of circulating lymphocytes was not affected by low dose CY. Moreover, flow cytometric analysis showed no significant changes in the percentage of circulating CD8+ or CD4+ T-cells or in the CD4/CD8 ratio at any time point after CY. Thus, administration of low dose CY to these patients caused impairment of nonspecific T-suppressor function without selective depletion of the CD8+ subset that is generally associated with that function. Several immunoregulatory models that are consistent with these observations are discussed.

Topics: Adult; Aged; Concanavalin A; Cyclophosphamide; Female; Flow Cytometry; Humans; Immune System; Lymphocyte Activation; Male; Middle Aged; Neoplasms; Phytohemagglutinins; T-Lymphocytes, Regulatory

We investigated the lysis of fresh human solid tumor cells by peripheral blood T lymphocytes in the presence of lectins and anti-CD3 monoclonal antibodies (mAb). Addition of certain lectins (Con A, PHA, or WGA) directly into the 4-hr 51Cr-release assay caused significant lysis of (P less than 0.001) noncultured solid tumor targets by enriched populations of granular lymphocytes (GL). Significant levels (P at least less than 0.001) of Con A- or PHA-dependent solid tumor lysis by GL-enriched lymphocytes were observed in 32 of 39 donors (82%) and 14 of 20 donors (70%), respectively. In contrast, the addition of other lectins (PNA, PWM, or LPS) or anti-CD3 mAb did not cause cytotoxicity. The levels of Con A-dependent lysis were comparable to those of interleukin 2 (IL-2)-induced lysis by Leu 11b+ natural killer (NK) cells. The presence of lectins at the effector phase, but not of recombinant IL-2 (rIL-2), was required for the lysis of solid tumor targets. Both Con A-dependent and rIL-2-induced lysis were totally inhibited by treatment of the effector cells with the lysosomotropic agent L-leucine methyl ester (LeuOMe). Effector cells responsible for Con A-dependent lysis of solid tumors expressed T3 (CD3), T8 (CD8), and Leu 7 antigens, but lacked T4 (CD4) and Leu 11 (CD16) antigens as determined by both negative and positive cell selection studies. Con A-dependent lysis was inhibited at the effector phase by anti-CD3 (OKT3 or anti-Leu 4) or anti-CD2 (OKT11) mAb. On the basis of their phenotype (Leu 7+ CD3+ CD8+ CD16-), we hypothesize that these effector cells may contain a population of cytotoxic T cells (CTL) generated in vivo against autologous modified cells that can lyse fresh solid tumor target cells under conditions where the recognition requirements for the CTL are bypassed by lectin approximation.

Topics: Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; Concanavalin A; Cytotoxicity, Immunologic; Humans; Immunity, Cellular; Interleukin-2; Killer Cells, Natural; Leucine; Neoplasms; Phytohemagglutinins; T-Lymphocytes

In crossed affino-immunoelectrophoresis pure haptoglobin or that present in serum moved in the shape of either non-retarded or weakly-, or strongly-retarded fractions, related to varying Con A concentrations included into the first dimension gel. At low concentrations of Con A non-, weakly-, and strongly-retarded fractions were observed, whereas at higher concentrations only one, strongly-retarded fraction could be found. The number and percentage composition of haptoglobin fractions were dependent not only on the concentration of Con A, but also on other proteins added to the sample submitted to electrophoresis. Irregular distribution of haptoglobin Con A-dependent fractions made quantitative measurements unreliable.

Topics: Concanavalin A; Haptoglobins; Humans; Immunoelectrophoresis, Two-Dimensional; Neoplasms

Topics: Concanavalin A; Histiocytes; Humans; Neoplasms; Staining and Labeling

Lymphocytes from 22 patients with established malignancy were stimulated with concanavalin A (Con A), and supernatants were tested for T-lymphocyte chemotactic factor (LCF). LCF activity was measured using a leading front chemotaxis assay with normal human T cells as responders. Fifteen of the 22 patients tested produced LCF at a level of less than 2 standard deviation below the mean of control cells. In 10 patients where mononuclear cells were stimulated with Con A for 24, 48, and 72 hr, LCF activity was significantly reduced at all three time points averaging 38, 14, and 43% of control levels, respectively. In 13 of the 31 patients, patient T-cell migration in response to casein was measured and compared to the production of LCF by mononuclear cells from these same patients. A significant correlation was observed indicating that both the response of T cells to a migration stimulus, and the production of T-cell-derived LCF was comparably suppressed. The reduction in LCF production by mononuclear cells from patients with established malignancy was not reversed by the addition of indomethacin to the culture system during Con A stimulation indicating that inhibition was not mediated by excessive prostaglandin production. The addition of patient mononuclear cells or T cells to normal mononuclear cells resulted in the inhibition of normal cell LCF by patient mononuclear cells or T cells. This could not be attributed to the production of a lymphocyte chemotactic inhibitor by patient cells, but appeared instead to be due to the direct inhibition of normal cell LCF synthesis or release by patient mononuclear cells or T cells. Separation of patient T cells into Leu 2 suppressor/cytotoxic or Leu 3 helper/inducer T cells showed that the inhibitory activity was associated with the Leu-2 T-cell subset. Heterologous normal Leu 2 T cells did not suppress normal mononuclear cell LCF production suggesting that patient Leu 2 T cells were functionally activated as compared to normal Leu 2 cells. The decreased production of LCF coupled with a depressed T-cell migratory activity in patients with established malignancy may in part be responsible for suppressed cellular immune reactions in these patients and possibly the impairment of tumor rejection.

Topics: Adolescent; Adult; Aged; Cell Separation; Chemotactic Factors; Chemotaxis, Leukocyte; Concanavalin A; Humans; Immunity, Cellular; Immunologic Deficiency Syndromes; Lymphocyte Activation; Middle Aged; Neoplasms; T-Lymphocytes; T-Lymphocytes, Regulatory

Most retroviruses are immunosuppressive in vitro and in vivo. They are able to enhance virus-induced tumor development and/or to induce acquired immune deficiency syndromes (AIDS) which are characterized by malignant tumors and opportunistic infections. Experimental evidence for the immunosuppressive properties of several type D viruses derived from human cell lines and other retroviruses is presented.

Topics: Acquired Immunodeficiency Syndrome; Animals; Cattle; Concanavalin A; Haplorhini; Humans; Immune Tolerance; Lymphocyte Activation; Mice; Neoplasms; Neoplasms, Experimental; Phytohemagglutinins; Retroviridae; Species Specificity

The immunological significance of the lymphocyte mobility histogram derived from a fully automated cell electrophoretic instrument (Parmoquant-L), was considered. In mouse lymphocytes, the ratio of medullary thymocytes increased as a result of a decrease in the proportion of cortical thymocytes, whereas the ratio of splenocytes in the low-mobility zone (0.85-0.95 micron/sec/V/cm) increased in the course of tumor growth or in vivo concanavalin A-treatment. In the peripheral lymphocytes of cancer patients, the decreased ratio of high-mobility T cells resulted in a gradual increase in that of low-mobility T cells, which were not seen in lymphocytes modified by the patient's serum. Although the formation of blastoid cells by stimulation of the tumor or mitogen cannot be completely denied, the low-mobility T cells of peripheral lymphoid tissues in the tumor-bearing status are considered to be caused by incomplete differentiation of the cells, in which immunosuppressor cells are involved. Pattern analysis of lymphocyte electrophoresis would be a simple method if fully automated instrumentation was used.

Topics: Animals; Cell Separation; Concanavalin A; Electrophoresis; Humans; Lymphocyte Activation; Mice; Mice, Inbred C3H; Mice, Nude; Neoplasms; Spleen; T-Lymphocytes

The percentage non-glycosylated, tissue type ferritin was measured in the serum of 30 cancer patients and 11 normal subjects, as well as in 10 tumour cytosols. As expected, in normal subjects the larger part of serum ferritin corresponded to glycosylated ferritin, and in tumour cytosols the larger part corresponded to non-glycosylated ferritin (tissue ferritin). In all but one cancer patients a significant part of the protein did not bind to Con A, thus behaving as tissue ferritin. The findings obtained suggest release of tissue type, non-glycosylated, ferritin by damaged cells or through abnormal secretion. This relative increase of the non-glycosylated ferritin fraction appears to be very frequently present in cancer, although not specific for it. Assaying for Con A binding may prove to be valuable in the study of malignant disease, and therefore deserves further investigation.

Topics: Absorption; Adult; Aged; Concanavalin A; Female; Ferritins; Humans; Kinetics; Male; Middle Aged; Neoplasms; Protein Binding; Transaminases

Major molecular species of human alpha-fetoprotein(AFP), which were separated as single components by serial affinity chromatography with concanavalin A(Con-A) and Lens culinaris agglutinin, were further resolved into several bands by affinity electrophoresis with erythroagglutinating phytohemagglutinin of Phaseolus vulgaris lectin(E-PHA). Among the newly separated main molecular species, both Con-A- and E-PHA-reactive AFP(AFP-1X1) was demonstrated, contrary to the known sugar specificity of Con-A and E-PHA, in addition to molecular species of AFP reacting with Con-A but not with E-PHA(AFP-1X0) and of AFP reacting with E-PHA but not with Con-A(AFP-0X1). AFP-0X1 was formed from AFP-0X0, and AFP-1X1 from AFP-1X0 by neuraminidase treatment; thus, AFP-0X1 and AFP-1X1 represent asialylated and AFP-0X0 and AFP-1X0 sialylated molecular species. AFP-1X1' and AFP-0X0' were present as minor components. AFP-0X0' had no affinity for E-PHA, and the affinity increased in the order of AFP's-0X0(or 0X1), -1X1', -1X1 and -0X1. Proportions of those components varied depending on the pathophysiological conditions of AFP production.

Topics: alpha-Fetoproteins; Asialoglycoproteins; Carcinoma, Hepatocellular; Chromatography, Affinity; Concanavalin A; Electrophoresis; Enzyme-Linked Immunosorbent Assay; Fetuins; Humans; Liver Neoplasms; Mesonephroma; Neoplasms; Neoplasms, Germ Cell and Embryonal; Phytohemagglutinins; Stomach Neoplasms; Substrate Specificity

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

Human thymic epithelial monolayer-conditioned medium (TEM-CM) enhanced concanavalin A (ConA)-induced suppressor T-lymphocyte activity in 15 of 17 studies of fractionated light-density bone marrow mononuclear cells (LD-BMMC) obtained from pediatric cancer patients within 7 days of chemotherapy (P less than 0.001). However, TEM-CM depressed ConA-induced suppressor T-lymphocyte activity in 14 of 18 studies of LD-BMMC obtained from patients who had received their chemotherapy 14-21 days previously (P less than 0.05). In studies of LD-BMMC from normal subjects, TEM-CM did not show any significant effect on suppressor cell activity, nor did TEM-CM significantly affect spontaneous suppressor cell activity in patients or normals. The effect of direct culture on thymic epithelial monolayers was equivalent to the effect of TEM-CM in both ConA-induced and spontaneous suppressor cell assays. These data demonstrate thymic factor-mediated changes in suppressor T-cell activity of pediatric cancer patients and suggest a postchemotherapy alteration in the bone marrow population of inducible prethymic T cells.

Topics: Adolescent; Antineoplastic Agents; Bone Marrow; Child; Child, Preschool; Concanavalin A; Culture Media; Humans; Neoplasms; T-Lymphocytes, Regulatory; Thymus Gland

The authors studied the effects of soluble syncytiotrophoblast extract (STE) on the proliferative responses of lymphocytes to different lectins (PHA and Con A) and to allogeneic cells in one-way mixed lymphocyte cultures. STE suppressed lymphocyte reactivity to lectins and to allogeneic cell cultures. The inhibitory effect was dose dependent. Increased concentrations of lectins failed to overcome the inhibitory effect of STE. Lymphocytes preincubated with STE for 18 hr, then washed and exposed to lectins still exhibited an inhibition of cellular proliferation. STE added to lymphocyte cultures at various times in the presence of both mitogens or of allogeneic cells continued to inhibit lymphoproliferative responses even when it was added after 43 hr of cell culture. Furthermore, STE was able to reduce the spontaneous proliferation of tumor cell lines K562 and LHN13 maintained in vitro culture prior to testing. In all cases, the inhibition observed was not due to lymphocytotoxicity or to tumor cell mortality. The inhibitory effect of STE on the proliferative responses of lymphocytes to stimulants may be due to a cytostatic effect that may represent a contributing factor in the nonrejection of the fetus by a competent immune system.

Topics: Cell Survival; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Immune Tolerance; In Vitro Techniques; Kinetics; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Neoplasms; Phytohemagglutinins; Pregnancy; Trophoblasts

Chronic alcoholics are more susceptible to infection and have increased incidences of certain types of carcinomas. One explanation for this may be suppressed immune responses secondary to ethyl alcohol consumption. This project was initiated to study the effect of ethyl alcohol on lymphocyte responses in vitro by monitoring tritiated thymidine uptake. Lymphocytes were incubated in the presence of phytohemagglutinin-P, concanavalin A, and pokeweed mitogen. The response of normal lymphocytes was noted after mitogen stimulation in the presence of ethyl alcohol in graded doses. Ethyl alcohol levels greater than or equal to 50 mg/dL suppressed tritiated thymidine uptake of normal lymphocytes for phytohemagglutinin-P and concanavalin A. Since ethyl alcohol exposure in concentrations consistent with blood levels that may be attained during routine ingestion significantly decreased lymphocyte blastogenesis, it is speculated that chronic ethyl alcohol ingestion may alter immune surveillance sufficiently to be responsible in part for the increased incidence of infection and/or neoplasms seen in alcoholic subjects.

Topics: Adult; Alcoholism; Concanavalin A; Dose-Response Relationship, Drug; Ethanol; Female; Humans; Infections; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Neoplasms; Phytohemagglutinins; Pokeweed Mitogens; Stimulation, Chemical; Thymidine

Serum ferritin has been suggested as a tumor marker in the diagnosis of certain malignancies and for following the activity or dissemination of the malignant process. Since neoplastic tissues generally contain more acidic isoferritins than their normal tissue counterparts, it has also been suggested that the specific assay of such isoferritins in serum may be of particular value in the diagnosis of malignancy. In this work, we have evaluated ferritin concentration in the serum of normal subjects and patients with acute nonlymphocytic leukemia, Hodgkin's disease, breast cancer and lung cancer by simultaneously using three different immunoassays: an immunoradiometric assay based on polyclonal antibodies against human liver (basic, L-subunit rich) ferritin, a radioimmunoassay based on polyclonad antibodies against HeLa cell (acidic, H-subunit rich) ferritin, and an immunoradiometric assay based on the monoclonal antibody 2A4 raised against human heart (acidic, H-subunit rich) ferritin. Most of the patients studied had increased values for liver-type ferritin in the absence of increased iron stores. Binding of serum ferritin to concanavalin A did not prove to be useful in distinguishing a tumor-specific basic isoferritin. The HeLa ferritin assay was found to be less specific than the heart ferritin assay in the detection of acidic isoferritins, and did not provide any advantage over the liver assay in detecting the increased levels of serum ferritin associated with malignant disease. Heart-type ferritin was found in one-fifth of normal sera and 64% of sera from patients with malignancy. Values were very low compared with those for basic ferritin, ranging from less than 0.1 to 17% of total serum ferritin (geometric mean value 1.3%) in patients with malignancy. These findings indicate that at present there is little application for serum ferritin immunoassays based on antibodies to HeLa cell or heart ferritin in the diagnosis or monitoring of malignant disease. This seems to be due to the presence in human serum of biding factors which are responsible for the rapid clearance of acidic isoferritins from the circulation. The serum concentration of basic ferritin, however, can be useful in the diagnosis and management of some malignancies, and it is possible that studies on cell isoferritins can be important in biologic monitoring of neoplastic disorders. It should also be noted that the increased levels of serum ferritin found in patients with malignancy can exer

Topics: Antibodies, Monoclonal; Concanavalin A; Ferritins; HeLa Cells; Humans; Immunoassay; Liver; Myocardium; Neoplasms

We have shown that cyclophosphamide (CY) can augment the development of delayed-type hypersensitivity to a primary antigen in patients with advanced cancer. In the present study, we administered CY (1000 mg/sq m) to 19 patients with advanced, metastatic cancer and monitored the compositional and functional changes in their peripheral blood mononuclear cells. Within 2 days of administration of CY, the lymphocyte count fell significantly (mean decrease = 26.0%) and remained significantly depressed through Day 14 with recovery beginning by Day 21. T- and B-lymphocytes were depleted to about the same degree at each time point. Moreover, there was no selective depletion of the Leu 2(+) (suppressor-cytotoxic) or Leu 3(+) (helper-inducer) subsets of T-lymphocytes. Proliferative responses to mitogens (phytohemagglutinin, concanavalin A, pokeweed mitogen) and to allogeneic cells fell significantly within 1 day of administration of CY and continued to be diminished on Day 2. However, these responses recovered to pretreatment levels by Day 3, and, in some cases, exceeded pretreatment levels on Day 7. Concanavalin A-inducible suppressor activity was also diminished on Day 1 (mean decrease, 23.4%) and Day 2 (mean decrease, 39.2%). However, in contrast to the proliferative responses, suppressor activity continued to be significantly impaired on Day 3 (mean decrease, 31.6%) and only partially recovered by Day 7 (mean decrease, 22.1%). Both concanavalin A-inducible suppression and proliferative responses declined again on Days 14 and 21. Thus, between 3 and 7 days after administration of CY, there appeared to be impairment of nonspecific T-cell-mediated suppressor activity of peripheral blood lymphocytes that was not merely a reflection of impaired lymphocyte function in general. This could account for the augmented delayed-type hypersensitivity responses of CY-treated patients.

Topics: Concanavalin A; Cyclophosphamide; Female; Humans; Lymphocyte Activation; Lymphocytes; Male; Monocytes; Neoplasms; T-Lymphocytes, Regulatory; Time Factors

The migration and concentration of lymphocytes at sites of antigenic challenge are an integral part of the expression of delayed cutaneous hypersensitivity, as well as of tumor and graft rejection. In this study, we have analyzed the migration of T lymphocytes from patients with malignancy. We used casein and concanavalin A (Con A)-stimulated mononuclear cell supernatants to stimulate T cell locomotion. Peripheral blood T lymphocytes from 30 patients with established malignancy, 10 patients with indolent malignancy or benign tumor, and 42 normal adult controls were tested. Data are expressed as a migration index (MI), which represents the difference in micrometers between the distance migrated in response to a stimulus and the distance migrated in response to media alone. We observed a marked depression in casein-stimulated T lymphocyte migration in patients with established malignancy (mean MI +/- 1 SD = 17.0 +/- 9 microns) as compared with normal adult controls (mean MI +/- 1 SD = 35.3 +/- 10 microns). Similar results were observed with migration in response to Con A supernatants. T cells from patients with established malignancy had a mean MI of 5.8 +/- 4 microns to Con A supernatants as compared with 24.5 +/- 5 for controls. This depressed migration was apparent both in the distance that cells migrated and in the number of cells that migrated into the membrane. Of 10 patients with indolent malignancy or benign tumor, T cell migration in 8 was not significantly decreased as compared with controls. When we mixed equal concentrations of normal control T lymphocytes with T lymphocytes from patients with cancer and added the mixture directly to the upper compartment of the chemotaxis chamber, the response of the normal T cells to casein was inhibited by an average of 48%. We observed inhibition of this migration of normal cells when we added as little as 10% of patient cells to normal cells. When we mixed normal control T lymphocytes from different donors and added them directly to the upper compartment of the chemotaxis chamber, T lymphocyte migration in response to casein was not significantly altered. If T cells from patients with cancer were cultured overnight, the suppressive effect on lymphocyte locomotion was lost. Our results indicate that there is a population of T lymphocytes in patients with cancer that suppress normal T lymphocyte migration. This suppressor activity may partially explain the subversion of immunosurveillance in established neopl

Topics: Adolescent; Adult; Aged; Caseins; Cell Movement; Chemotaxis, Leukocyte; Concanavalin A; Female; Humans; Immunity, Cellular; Lymphokines; Male; Middle Aged; Neoplasms; T-Lymphocytes; T-Lymphocytes, Regulatory

T cells from 19 out of 25 childhood cancer patients showed impaired proliferative responses to purified protein derivatives (PPD)-pulsed antigen-presenting cells (APC) although all of the patients had been immunized with BCG. To test whether such low responsiveness of T cells results from the dysfunction of T cells or from that of APC, the experiment was designed to assess the proliferative response of T cells from patients or their parents to PPD-pulsed APC from patients or parents. These combinations seem to be suitable to assess the activity of T cells or APC since at least partial identity of HLA-D/DR antigens is required for T cell-APC interactions. Although T cells from patients who showed low responsiveness to PPD failed to respond even to PPD-pulsed APC from parents, T cells from parents were able to respond to PPD-pulsed APC from patients as well as to autologous APC. These observations strongly suggest that the low responsiveness to PPD in childhood cancer patients results from the dysfunction of T cells, and the capacity of APC is fully preserved. In other words, it appears that the capacity of APC is not impaired by chemotherapy, neoplastic cells, or other factors. Suppressor T cells appeared not to be involved in such dysfunction of T cells.

Topics: Adolescent; Antigen-Presenting Cells; Child; Child, Preschool; Concanavalin A; Humans; Infant; Lymphocyte Activation; Neoplasms; Phytohemagglutinins; T-Lymphocytes; T-Lymphocytes, Regulatory

Topics: alpha 1-Antichymotrypsin; Chymotrypsin; Concanavalin A; Humans; Immunoelectrophoresis; Neoplasms

The lectin (PHA and Con A) induced agglutination of a human tumour cell line (lung adenocarcinoma and osteosarcoma) was estimated by the spectrophotometric method. A decrease in the optic density for 1 min (delta D) of cell suspension and the time (t0) necessary for a complete sedimentation of cells (at delta D = 0) were used as quantitative indicators of agglutination. An increase in the concentration of tumour cells and lectins resulted in an increase of delta D and a decrease of t0. The results of spectrophotometry were correlated with the microscopic study data for tumour cells agglutination.

Topics: Adenocarcinoma; Agglutination; Cell Aggregation; Cell Count; Cells, Cultured; Concanavalin A; Humans; Lectins; Lung Neoplasms; Neoplasms; Osteosarcoma; Phytohemagglutinins; Spectrophotometry

The monoclonal antibody 791T /36, prepared against a human osteogenic sarcoma cell line, 791T , reacts with a variety of human tumours and also mitogen-stimulated PBMN cells. The target antigen as expressed upon 791T cells is a monomeric plasma membrane-associated glycoprotein with an apparent Mr of 72000. By quantitative flow cytofluorimetry, approx. 10(5) antibody molecules bound per cell to T-lymphoblasts induced with PHA or Con A, whereas only a few thousand antibody molecules bound per cell to unstimulated cells, so that the antigen may be classified as a lymphocyte activation antigen. On lymphoblasts, the 791T /36 again reacted with a protein with an apparent Mr of 72000. This antigen therefore has a dual role as a tumour marker and lymphocyte activation antigen which may be implicated in the regulation of cell proliferation.

Topics: Antibodies, Monoclonal; Antigen-Antibody Complex; Antigens, Neoplasm; Antigens, Surface; Cell Line; Cells, Cultured; Concanavalin A; Flow Cytometry; Humans; Lymphocyte Activation; Neoplasms; Osteosarcoma; T-Lymphocytes

The in vitro effects of ISO on the immune responses of cancer patients were investigated. Forty seven patients with primary tumors (26 lung carcinoma, 14 breast adenocarcinoma, 7 melanoma) were studied. Concanavalin A (ConA)-induced lymphocyte proliferation, natural killer cell (NK) activity, and monocyte chemotaxis were measured. In 40 of the 47 patients (85%), ConA-induced lymphocyte proliferation was depressed; NK activity was depressed in 32 (68%), and monocyte chemotaxis was found to be depressed in 36 (77%). For in vitro studies, an optimum concentration of ISO (100 micrograms/ml per 10(6) cells) was used to treat peripheral blood mononuclear cells. In the presence of ISO, all three parameters were restored to normal or near normal levels in those that were depressed. Under these preincubation conditions in vitro treatment of mononuclear cells from the peripheral blood of normal individuals with ISO had no effect on their activities in the three assays. Similar effects on these three immune parameters were observed when 24 h supernatants obtained from patients' mononuclear cells pretreated with ISO were employed in these assays.

Topics: Adjuvants, Immunologic; Cell Division; Chemotaxis, Leukocyte; Concanavalin A; Humans; Immunity; Inosine; Inosine Pranobex; Killer Cells, Natural; Lymphocytes; Monocytes; Neoplasms; Time Factors

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

A sensitive method for human lectin-dependent cell-mediated cytotoxicity (LDCC) is presented using HEp-2 adherent human epipharynx carcinoma cells as targets. Cytotoxicity was evaluated by detachment from the monolayer of 3H-TdR-prelabelled HEp-2 cells. Maximal LDCC was obtained in a 24 h assay with a Con A dose of 25 micrograms/ml for 50 : 1 effector-target cell ratio requiring only 2500 target cells per well. Testing of five different lymphocyte fractions: peripheral blood mononuclear cells (PBMC), monocyte-enriched adherent cells (AC), monocyte-depleted non-adherent cells (non-AC), T and non-T lymphocytes as effector cells from 25 normal individuals, suggests that LDCC to HEp-2 targets is mediated by T lymphocytes.

Topics: Cells, Cultured; Concanavalin A; Cytotoxicity Tests, Immunologic; Cytotoxicity, Immunologic; Humans; Lectins; Lymphocytes; Neoplasms; T-Lymphocytes; T-Lymphocytes, Cytotoxic

The effects of Vitamin B compounds consisting of B1, B6 and B12 on the in vitro lymphocyte proliferative (LP) response were investigated. Vitamin B compounds failed to enhance the LP response of healthy controls but did enhance that of cancer patients, suggesting the recovery of depressed LP response to normal level by the treatment employing the compounds. The addition of Vitamin B compounds in the generation phase of Concanavalin-A induced suppressor cells significantly abrogated the activities. However, Vitamin B compounds produced no effect to neutralize the inhibitory effect of cancer sera on LP response.

Topics: Concanavalin A; Humans; In Vitro Techniques; Lymphocyte Activation; Lymphocytes; Neoplasms; Vitamin B Complex

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

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

Topics: Cell Membrane; Concanavalin A; Humans; Neoplasms; Staining and Labeling

Peripheral blood lymphocytes (PBL) from cancer patients were first tested for the proliferative response to concanavalin A (Con A). The lymphocytes which had lower response to Con A than the normal control were supposed to have relatively greater numbers of putative suppressor cells, or higher suppressive activity was anticipated in the PBL. The PBL were then treated with mitomycin C, added to normal lymphocytes in the presence of Con A, and cocultured for further investigation of the activity of the putative suppressor cells as determined by the effect of these putative cells on the responses of normal lymphocytes to Con A. In many of our studies inconsistent results showed between two types of assay systems. Not all patient's lymphocytes showing depression in response to Con A could suppress the proliferation of normal lymphocytes in response to Con A in coculture systems. However, some of the patients' lymphocytes, despite not showing a depressed response to Con A in the direct assay, were able to inhibit the response of normal lymphocytes to Con A in coculture. The contradictory data imply that it is inappropriate to conclude that suppressor cells are present at elevated levels in cancer patients by relying solely on the evidence of a depressed response to mitogens, either in a direct stimulation assay or in a coculture system. Our results possibly reflect that the development of cancer is not directly linked to the elevation of suppressor cell activity. Other more complicated mechanisms may be involved.

Topics: Concanavalin A; Humans; Lymphocyte Activation; Mitogens; Neoplasms; T-Lymphocytes, Regulatory

A glycoprotein was selectively enriched in the supernatant (Fraction b) obtained by alcohol and trichloroacetic acid fractionation of digitonin extracts from blood of patients with neoplastic diseases and of control subjects. Subsequent chromatography with concanavalin A:Sepharose separated a concanavalin A-reactive fraction from a concanavalin A-nonreactive one. In sodium dodecyl sulfate gel electrophoresis, the fractions from both malignant origin as well as control subjects appeared as single bands showing the same mobility. They were identical with the band obtained from commercial alpha 1-acid glycoprotein. In Fraction b of malignant origin, greatly increased amounts of the alpha 1-acid glycoprotein from malignant cases (AGPM) were found as compared to alpha 1-acid glycoprotein from controls (AGPC). Furthermore, AGPC had a higher glycine content than did AGPM. The electrofocusing pattern of AGPM showed additional bands between pH 3.7 and 4.4, whereas AGPC and commercial alpha 1-acid glycoprotein focused between pH 3.2 and 3.8. In contrast to AGPC and to a commercial alpha 1-acid glycoprotein, AGPM is characterized by a chromophoric group with maximal absorbance at 400 nm. It could be detached by treatment with 6 M guanidine hydrochloride thus indicating a noncovalent binding. The spectral data on the separated chromophore at pH 0.5 agreed with that of a 6,7-substituted pteridine. After detachment with reducing agents, a pteridine in its 7,8-dihydro form was indicated by spectral analysis.

Topics: Amino Acids; Carrier Proteins; Chromogenic Compounds; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Humans; Molecular Weight; Neoplasms; Orosomucoid; Pteridines

At temperatures above 37 degrees C, the abilities of many, but not of all, anticancer drugs to kill cells increases substantially. The increased cytotoxic rate can vary smoothly with temperature (e.g., nitrosoureas or cis-platinum(II) diamminedichloride) or show a marked threshold near 43 degrees C (e.g., adriamycin or bleomycin). Clinically, the increased cell killing rate can be used in the combination of localized hyperthermia and systemic chemotherapy. Drugs can also be useful in the study of cell killing by heat. Aliphatic alcohols which mimic and interact with heat, tend to be localized in lipid-rich regions of the cell, i.e., membranes. Heat modifies the rate at which plant lectins agglutinate cells and also modifies capping rates; most likely, these changes reflected membrane alterations. Because other data strongly imply that proteins are involved in cell death, we think that the first step in hyperthermic killing of cells involves damage to specific membrane-associated proteins.

Topics: Alcohols; Amphotericin B; Animals; Antineoplastic Agents; Cell Survival; Concanavalin A; Glucose; Hot Temperature; Humans; Neoplasms; Structure-Activity Relationship

A method is described for using a fluorescein isothiocyanate concanavalin A conjugate to stain human cell membranes in formalin fixed paraffin embedded tissue. 57 neoplastic and normal tissue sites were examined. In 54 malignant tumours, bright green fluorescence was confined to the cell membranes while in 23 benign tumours and normal tissue sites, the membranes were unstained or showed a diminished level of fluorescence. The distinction between malignant and hyperplastic or normal cells was clear cut and definite.

Topics: Breast Neoplasms; Concanavalin A; Female; Fluorescein-5-isothiocyanate; Fluoresceins; Fluorescent Antibody Technique; Humans; Liver Neoplasms; Lung Neoplasms; Neoplasms; Pancreatic Neoplasms; Skin; Skin Neoplasms; Thiocyanates

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: Adult; Aged; Concanavalin A; Female; Humans; Immunoglobulin G; Immunoglobulin M; Lymphocyte Activation; Macrophages; Male; Middle Aged; Neoplasms; Phytohemagglutinins; Receptors, Fc; T-Lymphocytes; T-Lymphocytes, Regulatory

Anergic patients with solid tumors were studied for the presence of suppressor cells which could be responsible for their significantly impaired phytohemagglutinin induced lymphocyte proliferation. Three types of suppressor cell assays were measured. First, no increased activity of prostaglandin producing suppressor cells was found in 10 cancer patients. Second, 5 cancer patients did not have significantly increased activity of short-lived suppressor cells in comparison with normal subjects. Third, no enhanced activity of concanavalin A induced suppressor cells was seen in 6 patients. This study emphasizes that the in vitro T cell immunodeficiency in this group of cancer patients was not due to suppressor cells as measured by conventional in vitro assays and emphasizes the potential importance of a loss of responder cells or other suppressor mechanisms in immune deficient patients with solid tumors.

Topics: Adult; Concanavalin A; Female; Humans; Immune Tolerance; Indomethacin; Leukocyte Count; Lymphocyte Activation; Male; Middle Aged; Neoplasms; T-Lymphocytes, Regulatory

Topics: Adolescent; Adult; Cells, Cultured; Concanavalin A; Humans; Neoplasms; Phytohemagglutinins; T-Lymphocytes, Regulatory

Topics: Agammaglobulinemia; Animals; Autoimmune Diseases; Chickens; Concanavalin A; Humans; Immunologic Deficiency Syndromes; Lymphocyte Cooperation; Macrophages; Mice; Mycoses; Neoplasms; Prostaglandins; T-Lymphocytes; T-Lymphocytes, Regulatory

The lymphocyte costimulator (CoS) is a lymphokine required for the activation of T cell responses to H-2 alloantigens or mitogen, CoS activity is found in the supernatant medium of Concanavalin A (Con A) stimulated spleen cells, In this paper we investigate the cellular requirements for CoS production by Con A-activated mouse spleen cells. Maximal lymphokine production in response to Con A depends on a co-operative interaction between T cells and a nylon wool-adherent cell present in the spleen of nude mice. T cells appear to be the major producers of CoS activity, doing so only in response to an initial inductive stimulus supplied by nude spleen cells. The inductive stimulus is found as a soluble factor in the supernatant of Con A-activated spleen cells, and can also be provided by stimulatory (S+), but not by non-stimulatory (S-), tumour cells H-2 identical with the responding T cells. The activation of lymphokine-producing T cells is thus a two-signal process, requiring both mitogen and an additional inductive signal. Once activated, homogeneous populations of T cells will release lymphokine in response to mitogen alone.

Topics: Animals; Cells, Cultured; Concanavalin A; Female; Interleukin-1; Lymphocytes; Lymphokines; Male; Mice; Neoplasms; Protein Biosynthesis; Spleen

We studied subcellular localization of saccharide moieties in cultured normal and malignant cells fixed in paraformaldehyde and treated with a nonionic detergent, using lectins specific for various surgar residues as probes in fluorescence microscopy. In normal cells, concanavalin A and Lens culinaris agglutinin, specific for mannose-rich carbohydrate cores in glycoproteins, labeled the endoplasmic reticulum as a wide perinuclear region. Other lectins, on the other hand, stained the Golgi apparatus as a juxtanuclear reticular structure. A similar compartmentalization was also seen in all malignant cells studied, although the Golgi apparatus in these cells was distinctly vesicular in appearance. Our results indicate that saccharide moieties in both normal and malignant cells are similarly compartmentalized, and thus speak in favor of a unidirectional subcellular flow for both membrane and secreted glycoconjugates.

Topics: Animals; Carbohydrates; Cell Compartmentation; Cell Line; Concanavalin A; Dogs; Endoplasmic Reticulum; Golgi Apparatus; HeLa Cells; Humans; Lectins; Neoplasms; Puromycin; Rats; Vinblastine

Topics: Concanavalin A; Dose-Response Relationship, Immunologic; Graft vs Host Reaction; Humans; Lymphocyte Activation; Neoplasms; Phytohemagglutinins; T-Lymphocytes, Regulatory; Thymus Hormones

Topics: Aging; Animals; Concanavalin A; Cyclic AMP; Cyclic GMP; Kinetics; Male; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Neoplasms; Spleen

Cytotoxic effector lymphocytes were induced in cultures of mouse spleen or lymph node cells by lymphocytosis promoting factor (LPF). The LPF-activated cytotoxic cells: (a) were not generated unless proliferation occurred; (b) sedimented in the lighter density fraction of a bovine serum albumin gradient; (c) were large, blast-like cells; and (d) were lysed by Thy-1.2 antiserum plus complement and, therefore, were T cells. Neither LPF alone nor supernates from stimulated cultures were cytotoxic. Unlike the situation with concanavalin A and phytohemagglutinin P, LPF-stimulated cytotoxic effector lymphocytes required no further addition of mitogen for maximal cytotoxicity. The effector cells displayed specificity, destroying only allogeneic but not syngeneic normal cells; in the case of tumor cells, both allogeneic and syngeneic cells werelysed in the absence of added mitogen. The reason for differentiated cytotoxicity toward syngeneic tumor and normal cells is not clear but may have some relevance to in vivo tumor rejection initiated by Bordetella pertussis.

Topics: Animals; Antilymphocyte Serum; Bordetella pertussis; Cell Division; Concanavalin A; Cytotoxicity, Immunologic; Female; Lymphocyte Activation; Lymphocytosis; Mice; Mice, Inbred Strains; Mitomycins; Neoplasms; Phytohemagglutinins; T-Lymphocytes

Topics: Animals; Cell Adhesion; Chemotactic Factors; Concanavalin A; Inflammation; Macrophage Migration-Inhibitory Factors; Macrophages; Male; Mice; Mice, Inbred C3H; Neoplasms; Peptones; Phagocytosis; Phytohemagglutinins

Topics: Carbohydrates; Chorionic Gonadotropin; Chromatography, Affinity; Colon; Concanavalin A; Female; Humans; Liver; Lung; Male; Neoplasms; Placenta; Pregnancy; Radioimmunoassay; Radioligand Assay

A variation was noted between normal subjects and cancer patients in the response of their lymphocytes to different antigens. Raising the temperature from 37 degrees C to 40 degrees C aided in improving lymphocytic activity to the antigenic stimulation. The heat response was more noticeable in normal subjects than in cancer patients, which suggests that immunotherapy should be initiated early in patients with malignant disease before there is systemic involvement from an increasing tumor burden.

Topics: Bacterial Toxins; Concanavalin A; Hot Temperature; Humans; Immunotherapy; Lymphocyte Activation; Neoplasms; Phytohemagglutinins; Tuberculin

Topics: Concanavalin A; Humans; Lectins; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Neoplasms; Rosette Formation

The abnormal membrane glycoprotein that we previously found to be associated with malignancy in a wide range of different tumours is present in the cell membrane as a dimer and appears to have a specific binding site for glucose.

Topics: Animals; Cell Membrane; Concanavalin A; Electrophoresis; Glucose; Glycoproteins; Humans; Lectins; Membrane Proteins; Neoplasm Proteins; Neoplasms; Protein Binding; Rats

Cell-to-cell interaction was investigated in various malignant tumor cells (human ovarial tumor, lung cancer, carcinoma of larynx and hamster melanoma cell) and in human lymphoblastoid cells (T-cell (MOLT-4 cell), thymoma cells and B-cells (Burkitt lymphoma cell)). Live lymphoblastoid cells did not adhere to the cell surfaces of tumor cells nor the lymphoblastoid cells were ingested by tumor cells without immunologic and specific treatment. Tumor cells as well as T-cells and B-cells had receptors to concanavalin A on their surfaces, and they showed marked cell binding of tumor cells and lymphoblastoid cells. Moreover, tumor cells that phagocytized lymphoblasts underwent marked cell destruction within 4 hours of cell binding. The cytolytic mechanism of the target tumor cell was probably related to contact with the lymphoblastoid cells and was increased by ingestive activity, and metabolic disturbance by lymphotoxin in tumor cells.

Topics: Animals; B-Lymphocytes; Cell Line; Concanavalin A; Cricetinae; Cytotoxicity, Immunologic; Humans; Lymphocytes; Neoplasms; Phagocytosis; Rosette Formation; T-Lymphocytes

Concanavalin A and wheat germ agglutinin, lectins that interact with serum glycoprotein in a manner similar to the antigen--antibody reaction, were used as "antibodies" in a single radial immunodiffusion technique to test a coded serum panel (from the National Cancer Institute, Bethesda, Md., and the Mayo Clinic, Rochester, Minn.) containing a) 99 serum samples from patients with different types of malignant neoplasms of the gastrointestinal tract, prostate gland, and lung, b) 50 samples from patients with benign diseases of the same organs as those affected in the cancer patients, and c) 50 samples from apparently healthy smokers. The resulting precipitation rings were not correlated to serum protein concentration, and the differences (demonstrated by Student's t-test and with a generalization of the one-sided two-sample Kolmogorov-Smirnov statistic for evaluating diagnostic tests) established that serum glycoproteins are glycosylated differently in cancer patients than in people without cancer.

Topics: Blood Proteins; Concanavalin A; Female; Gastrointestinal Neoplasms; Glycoproteins; Humans; Immunodiffusion; Lectins; Lung Neoplasms; Male; Neoplasm Proteins; Neoplasms; Prostatic Neoplasms

The reactivity of peripheral blood lymphocytes from patients with advanced malignancy was assessed by mitogen-induced stimulation of protein synthesis as measured by 3H-leucine incorporation. It was confirmed that the lymphocyte response of patients was depressed. Furthermore, the lymphocytes of 15 out of 27 cancer patients, selected because of their low responses, inhibited the reactivity of normal lymphocytes in co-cultures. The lymphocytes from one patient with Hodgkin's disease were also inhibitory. In contrast, lymphocytes from healthy subjects, patients with chronic lymphocytic leukaemia, lymphosarcoma or multiple myeloma caused no suppression. Experiments with purified cell populations from patients with carcinoma indicated that purified T cells responded to mitogens while unseparated lymphocytes failed to respond and that the inhibitory activity was due to adherent cells, presumably monocytes. There was no evidence for B-cell-mediated suppression. However, in two cases inhibition was caused by isolated T cells of the patients and not by adherent cells. These experiments suggested that one mechanism for the depression of cell-mediated immunity seen in patients with advanced cancer may be the nonspecific suppresssion of certain T-cell functions by circulating monocytes.

Topics: Adult; Aged; Carcinoma, Bronchogenic; Cells, Cultured; Concanavalin A; Hodgkin Disease; Humans; Immune Adherence Reaction; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocyte Transfusion; Lymphocytes; Lymphoma, Non-Hodgkin; Middle Aged; Mitogens; Multiple Myeloma; Neoplasm Proteins; Neoplasms; T-Lymphocytes

The virus plaque assay (VPA), an assay capable of enumerating mitogen- or antigen-sensitive T-lymphocytes in a given cell population, was applied to the study of mitogen responses of peripheral blood leukocytes (PBL) in 50 patients with localized, solid cancers and in 29 normal controls. Concanavalin A (Con A)-responsive virus plaque-forming cells (V-PFC) were significantly reduced in the patients as compared with those of the controls. PBL from 20 of the patients but only 2 of the controls failed to show significant increments in V-PFC over background when cultured in the presence of Con A. This deficient response was also present in the patients when phytohemagglutinin was used as the mitogenic agent. Mitogen-stimulated uptake of [3H]thymidine (TdR) in parallel cultures failed to show a statistically significant difference between the patients and the controls, though some patients showed diminished stimulation in this assay. The VPA thus appeared to detect a defect of T-lymphocyte function not found in the [3H]TdR incorporation assay.

Topics: Cell Separation; Concanavalin A; Female; Humans; In Vitro Techniques; Lectins; Lymphocyte Activation; Male; Methods; Neoplasms; T-Lymphocytes; Thymidine

Topics: Aged; Concanavalin A; Humans; Lectins; Lymphocyte Activation; Middle Aged; Neoplasms; Rosette Formation; Skin Tests; T-Lymphocytes

The effect of operation on in vitro lymphocyte function in 35 cancer patients was studied. Lymphocyte proliferative responses to phytohemagglutinin (PHA), poke-weed mitogen (PWM), and concanavalin A (Con A) were measured by 3H-thymidine incorporation. Sheep red blood cell (SRBC) rosette formation also was quantitated. These tests were performed preoperatively and at 24 hours, one week, and 4 weeks postoperatively. Intra-abdominal and intrathoracic procedures, transfusions, and longer operating times depressed the lymphocyte proliferative response. However, an increased lymphocyte proliferative response was noted in sarcoma patients 24 hours postoperatively, possibly as a result of lowered tumor burden. Several of these changes still were evident 4 weeks after operation. Rosette formation also decreased significantly 24 hours postoperatively in patients who had intrathoracic or intra-abdominal procedures, but returned to preoperative levels after one week. In general, operation appears to cause transient depression of lymphocyte function in some cancer patients. However, lymphocyte function returns to normal by the fourth postoperative week in most patients.

Topics: Concanavalin A; Humans; Immune Adherence Reaction; Immunity; Immunosuppression Therapy; Lectins; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Middle Aged; Neoplasms

Within the last decade a variety of techniques have been developed and used for the detection of cell-mediated immunity in man by means of leucocyte migration inhibition in vitro. A detailed description of the leucocyte migration capillary tube technique (LMCT) and the leucocyte migration agarose technique (LMAT) is given. The procedure for selecting and using the proper antigen concentration is described. A description is also given of the indirect LMAT and the technique for determination of concanavalin A-induced lymphocyte release of leucocyte migration inhibition factor. Applications of these techniques are mainly intended for the exploration of the immunobiology of lymphocytes and cellular interactions associated with the immune response and the investigation of clinical conditions in man, i.e. infectious diseases, autoimmune diseases, transplantation states, tumour diseases, contact hypersensitivity and immunological deficiency states. Selection and adaptation to suit the experimental aim is necessary to obtain optimal results with these techniques. Their usefulness may be increased through more extensive use of purified antigens and indirect assays.

Topics: Cell Migration Inhibition; Concanavalin A; Humans; Immunity, Cellular; Lymphocyte Activation; Lymphocytes; Macrophage Migration-Inhibitory Factors; Neoplasms

The effect of thymosin on in vitro reactivity of peripheral lymphocytes to phytohemagglutinin, concanavalin A, and the formation of spontaneous E-rosettes in 54 patients with metastatic carcinomas has been studied. Thymosin increased lymphocyte responses to PHA and Con A in only 10 patients, with predominant effect seen with Con A. Twenty patients showed depressed baseline levels of E-rosettes which were increased to normal or subnormal levels after incubation with thymosin. No distinct correlation was noted between the clinical stage of the disease and the ability of lymphocytes to form E-rosettes. Although the exact mechanism by which the thymus exerts its influence on host resistance is not clearly defined, present evidence supports the concept that the thymic hormone, thymosin, may be an important addition in treatment of cancer patients by increasing cell-mediated immunity.

Topics: Adolescent; Adult; Aged; Breast Neoplasms; Carcinoma, Bronchogenic; Concanavalin A; Hodgkin Disease; Humans; Immune Adherence Reaction; In Vitro Techniques; Lectins; Middle Aged; Neoplasms; T-Lymphocytes; Thymosin; Thymus Extracts

The blastogenic response of lymphocytes from patients with malignant neoplasms was evaluated by stimulation with three phytomitogens (PHA, PWM, and Con A). The response of patient lymphocytes to all three mitogens was significantly lower than that of control lymphocytes, and most patients with abnormal PHA responses also responded abnormally to PWM and Con A. However, a few patients with normal PHA responses were abnormal to Con A, suggesting the suppression of a Con A-sensitive population. The observation that PWM responses were abnormal in patients with lowered PHA lymphocyte stimulation indicates that both T and B lymphocyte mitogen responses were suppressed in these patients. Plasma from patients was capable of either inhibiting or enhancing lymphocyte mitogen stimulation. However, inhibitory plasmas were generally from patients with abnormal mitogen responses.

Topics: Concanavalin A; Humans; In Vitro Techniques; Lectins; Lymphocyte Activation; Neoplasms

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

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

Lymphocytes were isolated from the peripheral blood of 21 normal persons and 66 patients with chronic lymphocytic leukemia (CLL), CLL in remission, Hodgkin's disease, Hodgkin's disease in remission, various other tumors, or cardiovascular diseases; The lymphocytes were studied for cap formation and agglutinability by concanavalin A, and for cell attachment to the surface of a petri dish. The frequency of cap formation was lowest in lymphocytes from patients with untreated Hodgkin's disease (2.1 plus or minus 0.8%), next lowest in lymphocytes from patients with CLL who were or were not under treatment (7,0 plus or minus 1;3%), and also low in Hodgkin's disease in remission (10.6 plus or minus 1.2%). The frequencies of cap formation by lymphocytes from patients with various other tumors (19.1 plus or minus 2.5%), with CLL in remission (24.0 plus or minus 0.9%), and with nonmalignant diseases (26.0 plus or minus 2.2%) were more similar to the frequency found in lymphocytes from normal persons (29.4 plus or minus 2.8%). Lymphocytes from all the patients, including those in remission, showed a higher degree of agglutinability by concanavalin A than lymphocytes from normal persons. Cell attachment to a petri dish was highest with CLL, next highest with CLL in remission, and low for normal persons and all the other patients. Lymphocytes from normal persons that consisted predominantly of thymus-derived cells gave similar results to isolated normal bone marrow-derived cells. The results indicate that there were different changes in the surface membrane of lymphocytes from patients with CLL, CLL in remission, Hodgkin's disease, and Hodgkin's disease in remission, and that the patients in clinical remission still showed abnormalities in their lymphocytes.

Topics: Adolescent; Adult; Aged; Agglutination; Cell Adhesion; Cell Membrane; Concanavalin A; Female; Hodgkin Disease; Humans; Leukemia, Lymphoid; Lymphocytes; Male; Middle Aged; Neoplasms; Remission, Spontaneous

Blocking factors are small polypeptide molecules that may appear in the serum of patients with cancer. These factors block the transformation of lymphocytes in culture to nonspecific mitogens such as phytohemagglutinin or concanavalin A and, therefore, may reflect changes in the immunocompetence of the patient. Blocking factors were monitored during the clinical course of thirty-five patients with cancer. These factors did not develop in patients with response to therapy whereas they did develop in patients without response. A third group of patients without response to therapy after a previous remission showed an absence of lymphocyte responsiveness in culture that was not due to blocking factors, suggesting that immune clone consumption had occurred. Dermal responsiveness to tumor antigen correlated with a favorable clinical course and was usually absent when serum blocking factors were present.

Topics: Antibody Formation; Antigens, Neoplasm; Carcinoma, Squamous Cell; Concanavalin A; Humans; Immunotherapy; Lectins; Leiomyosarcoma; Lymphocyte Activation; Lymphocytes; Melanoma; Neoplasms; Osteosarcoma; Peptides; Pharyngeal Neoplasms; Rhabdomyosarcoma; Skin Tests

Spleen cells cultured with syngeneic trinitrophenyl (TNP)-modified stimulator cells display a cytotoxic effect against syngeneic TNP-modified targets, but not against modified targets from unrelated H-2 haplotypes. Targets that share the K and I region of the H-2 complex with the stimulator (or effector) cell are lysed to the same extent as the specific targets, while targets that share the I region only are not. When only the D region is shared, a weak cytotoxic effect is observed. Therefore, the stimulator (or effector) and target cell must share the K or D but not the I region of the H-2 complex in order for optimal cytotoxicity to occur. Spleen cells sensitized to irradiated TNP-modified H-2-allogeneic cells are cytotoxic to these specific cells. Coculture of F1 hybrid cells with irradiated TNP-modified parental cells result in a cytotoxic effect against only those specific parental cells and not TNP-modified cells from the other parent. The cytotoxic effect of the F1 effector cells in the cell-mediated lympholysis test is blocked by the addition of unlabeled TNP-modified targets that are H-2 syngeneic with the sensitizing parental strain, but not H-2 syngeneic with the other parental strain. These data demonstrate that the specificity of the effector cell in this syngeneic cytotoxicity system is directed against altered self H-2-controlled-gene products, rather than a requirement for sharing of histocompatibility genes between effector and target cell in order for lysis to occur. The role of H-2 antigens in determining the sensitivity of a target cell to T-cell-mediated lysis is discussed.

Topics: Animals; Cesium Radioisotopes; Chromium Radioisotopes; Chromosome Mapping; Concanavalin A; Cross Reactions; Cytotoxicity Tests, Immunologic; Gamma Rays; Histocompatibility Antigens; Hybridization, Genetic; Lymphocytes; Mice; Mice, Inbred Strains; Neoplasm Transplantation; Neoplasms; Nitrobenzenes; Radiation Effects; Spleen; T-Lymphocytes; Trinitrobenzenesulfonic Acid

Agglutination by two lectins, Concanavalin A (Con A) and Ricinus communis agglutinin (RCA), has been investigated in a human lymphoid cell system. The main conclusions of this study are: (1) no systematic correlation exists between the neoplastic state and sensitivity to Con A or RCA; (2) cells of neoplastic lines vary unsystematically in their surface properties as evaluated by Con A agglutination, with the possible exception that presence of Epstein-Barr virus (EBV) is associated with a high degree of agglutination and (3) cells of diploid lymphoblastoid lines and phytohemagglutinin (PHA)-stimulated lymphoctes agglutinate similarly and significantly better than unstimulated T- or B-lymphocytes. The relatively simple Con A agglutination assay can be used as an adjunct in classification of human lymphoid cell lines.

Topics: Agglutination; Cell Line; Concanavalin A; Humans; Hyaluronoglucosaminidase; Lectins; Leukemia; Lymphocyte Activation; Lymphocytes; Lymphoma; Multiple Myeloma; Neoplasms; Neuraminidase; Plant Lectins; Plants, Toxic; Ricinus; Stimulation, Chemical; Trypsin

Topics: Animals; Antigens, Neoplasm; Antineoplastic Agents; Bleomycin; Carmustine; Cells, Cultured; Chickens; Clone Cells; Concanavalin A; Culture Techniques; Cytarabine; Cytological Techniques; Drug Resistance; Drug Synergism; Drug Therapy, Combination; Humans; Immunity, Cellular; Immunotherapy; Lectins; Lipopolysaccharides; Lymphocytes; Melanoma; Mice; Mitosis; Neoplasms; Neoplasms, Experimental; Thymidine

Topics: Adenocarcinoma; Concanavalin A; Humans; Hypersensitivity, Delayed; Immunologic Memory; Lectins; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Melanoma; Neoplasms; Nitrobenzenes; Sarcoma; Skin Tests

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

Topics: Animals; Antibodies, Neoplasm; Antibody Formation; Antigen-Antibody Reactions; Antigens, Neoplasm; Antilymphocyte Serum; Autoantibodies; Concanavalin A; Cytotoxicity Tests, Immunologic; Fibrosarcoma; Histocompatibility Antigens; Humans; Immunotherapy; Lymphocytes; Mammary Neoplasms, Experimental; Melanoma; Methylcholanthrene; Mice; Neoplasm Transplantation; Neoplasms; Neuraminidase; Rats; Sarcoma, Experimental; Vibrio cholerae

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

The in vitro response of hamster lymphocytes to concanavalin A (Con A) was investigated by using the (3)H-thymidine incorporation assay, and the results obtained were compared to those with phytohemagglutinin (PHA-P). The optimum culture conditions for stimulation were obtained when one million lymph node cells were cultivated for 72 hr in 2 ml of RPMI-1640 medium supplemented with 10% heat-inactivated fetal calf serum in the presence of either 2 mug of Con A or 0.05 ml of PHA-P. The average ratios of stimulation using lymph node cells with Con A and PHA-P were 227 and 14, respectively. Bone marrow and thymus cells responded very poorly or not at all. Methyl alpha-d-glucopyranoside, when added to the culture medium at the time of addition of Con A, completely inhibited stimulation. The lymphocyte stimulation could be reversed by addition of the sugar as late as 18 hr after the addition of Con A. The lymphocyte response from hamsters bearing tumors induced by simian virus 40 tumor cells and from hamsters immune to tumor transplants was comparable to the response of lymphocytes from healthy donors.

Topics: Animals; Bone Marrow; Cells, Cultured; Concanavalin A; Cricetinae; Culture Media; Glucose; Immunity, Cellular; Lectins; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Neoplasms; Spleen; Stimulation, Chemical; Thymidine; Thymus Gland; Time Factors; Tritium

Topics: Agglutination; Amino Acids; Animals; Blood Group Antigens; Carbohydrate Metabolism; Carbohydrates; Cells, Cultured; Chemical Precipitation; Concanavalin A; Glycoproteins; Hemagglutination; Lectins; Lymphocytes; Mice; Mitogens; Neoplasms; Plant Proteins; Polysaccharides; Protein Binding

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

Topics: Acetylcholine; Adult; Brain; Brain Chemistry; Breast Neoplasms; Bronchial Neoplasms; Carcinoma; Cell Migration Inhibition; Concanavalin A; Epinephrine; Epitopes; Female; Histamine; Humans; Laryngeal Neoplasms; Lymphocytes; Macrophages; Male; Middle Aged; Myasthenia Gravis; Neoplasm Proteins; Neoplasms; Nerve Tissue Proteins; Norepinephrine; Sarcoidosis; Serotonin; Stomach Neoplasms; Succinylcholine; Tryptamines

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