concanavalin-a and Leukemia

A case of chronic lymphoproliferative disorder is presented, wherein a morphologically homogeneous population of lymphoid cells displayed properties similar to those described for large granular lymphocytes (LGL). Besides their LGL-like phenotype (VEP 13+, OKM 1+, OKT 10+ Fc-IgG-receptor+, OKT 3-), the proliferating cells were cytotoxic to NK targets as well as to antibody-coated target cells. Clinically, our patient presented low-grade lymphocytosis, splenomegaly, neutropenia, hyperimmunoglobulinemia and recurrent infections. Based upon this and 32 similar cases reported in the literature, we conclude that lympho-proliferative disorders involving GL encompass a variety of clinical entities, ranging from reactive GL lymphocytoses to overt lymphocytic malignancies.

Topics: Adult; Aged; Antibodies, Monoclonal; Bone Marrow; Concanavalin A; Female; Humans; Hypergammaglobulinemia; Isoantigens; Killer Cells, Natural; Leukemia; Lymphocyte Activation; Lymphocytes; Lymphocytosis; Male; Microscopy, Electron; Middle Aged; Neutropenia; Phenotype; Phytohemagglutinins; Receptors, Fc; Receptors, IgG

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; 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

We developed a new electrochemiluminescence (ECL) platform for ultrasensitive and selective detection of leukemia cells. In order to construct the platform, the nonporous gold with controllable three-dimensional porosity and good conductivity was used to modify the screen-printed carbon electrode. The carbon quantum dots (CQDs) coated ZnO nanosphere (ZnO@CQDs) were used as good ECL label with low cytotoxicity and good biocompatibility. Structure characterization was obtained by means of transmission electron microscopy and scanning electron microscopy images. The aptamer was used for cell capture and the concanavalin A conjugated ZnO@CQDs was used for selective recognition of the cell surface carbohydrate. The proposed method showed a good analytical performance for the detection of K562 cells ranging from 1.0 × 10(2) to 2.0 × 10(7) cells mL(-1) with a detection limit of 46 cells mL(-1). The as-proposed device has the advantages of high sensitivity, nice specificity and good stability and could offer great promise for sensitive detection of leukemia cells in response to therapy.

Topics: Aptamers, Nucleotide; Biosensing Techniques; Carbon; Cell Separation; Concanavalin A; Equipment Design; Humans; K562 Cells; Leukemia; Luminescent Measurements; Quantum Dots; Sensitivity and Specificity; Zinc Oxide

Ganoderma lucidum (G. lucidum) is a medicinal mushroom having biological effects such as immunomodulation and anti-tumor actions. In China and many other Asian countries, G. lucidum is used as a folk remedy to promote health and longevity. Although many studies have shown that G. lucidum modulates the immune system, including, for example, antigen-presenting cells, natural killer (NK) cells, and the T and B lymphocytes, the effects of G. lucidum on the WEHI-3 leukemic BALB/c mice are unclear. We attempted to determine whether G. lucidum would promote immune responses in BALB/c mice injected with WEHI-3 leukemia cells. The effects of G. lucidum on the survival rate of WEHI-3 leukemia cells injected into BALB/c mice were examined. It increased the percentages of CD3 and CD19, but decreased the percentages of Mac-3 and CD11b markers, suggesting that differentiation of the precursor of T and B cells was promoted but macrophages were inhibited. It decreased the weight of spleens as compared with control mice. It also promoted phagocytosis by macrophage from peripheral blood mononuclear cell (PBMC) and it also promoted natural killer cell activity. It decreased the percentage of leukemia cells in the spleens of mice before they were injected with WEHI-3 cells. Apparently, G. lucidum affects murine leukemia WEHI-3 cells in vivo.

Topics: Animals; Biomarkers; Cell Extracts; Cell Line, Tumor; Cell Proliferation; Cell Survival; Concanavalin A; Injections; Killer Cells, Natural; Leukemia; Leukocytes; Liver; Macrophages; Male; Mice; Mice, Inbred BALB C; Phagocytosis; Reishi; Spleen; Survival Analysis; Time Factors; Xenograft Model Antitumor Assays

Measurements of fluorescence resonance energy transfer efficiency (E), via fluorescence polarization, have been applied to distinguish between chronic lymphocytic leukemia (CLL) patients and healthy persons. Capping of Concanavalin-A receptors is more pronounced in normal lymphocytes than in those of CLL patients. Membrane capping decreases the distance between donor and acceptor molecules embedded in the membrane (r), and thus increases the monitored energy transfer efficiency (E approximately 1/r6). Blood samples of 10 healthy subjects and 16 CLL patients were examined. In the healthy subjects, the mean E value for capped lymphocytes was 19 +/- 3, whereas in the CLL patients it was significantly lower (8 +/- 5) (P < 0.01).

Topics: Concanavalin A; Female; Fluorescence Polarization; Fluorescence Resonance Energy Transfer; Humans; Leukemia; Lymphocytes; Male; Phenotype; Random Allocation

Using the ratiometric Ca2+ indicator, indo-1, the antigen-induced increase in intracellular Ca2+ concentration ([Ca2+]i) was measured in individual RBL-2H3 cells which had been passively sensitized with monoclonal antibody to the dintrophenyl (DNP) haptenic group. Antigenic stimulation using DNP-human serum albumin conjugate (DNP-HSA) induced concentration-dependent asynchronous Ca2+ oscillations, or irregular spikes. To achieve a quantitative comparison of the effects of different concentrations of antigen on changes in Ca2+[i, the area under the curve (AUC) of Ca2+ oscillations in each cell was calculated. The dose-response curve of the calculated AUC is consistent with the bell-shaped dose-response curve for antigen-induced mediator release, depolarization and 86Rb(+)-efflux. Ca2+ oscillations induced by antigenic stimulation were abolished by removal of external Ca2+ and the subsequent reintroduction of external Ca2+ caused their resumption. To investigate the role of Ca2+ oscillations in the secretory response, changes in [Ca2+]i induced by concanavalin A (Con-A), A23187, thapsigargin and NECA were also monitored. Con-A mimicked the response induced by antigen, whilst A23187 and thapsigargin induced a large transient non-oscillatory response. NECA, an adenosine receptor agonist, induced only a small transient rise in Ca2+[i without oscillatory behaviour. Since all these stimuli accept NECA-induced degranulation in these cells, it is suggested that, although Ca2+ oscillations are not essential for the initiation of secretion, they probably underlie the in-vivo physiological response of mast cells and basophils to an antigenic challenge. They also seem to enhance the efficacy of the Ca2+ signal.

Topics: Adenosine-5'-(N-ethylcarboxamide); Animals; Antigens; beta-N-Acetylhexosaminidases; Calcimycin; Calcium Signaling; Cell Polarity; Concanavalin A; Dinitrophenols; Dose-Response Relationship, Drug; Ionophores; Leukemia; Membrane Potentials; Radioisotopes; Rats; Receptors, IgE; Rubidium; Serum Albumin; Tumor Cells, Cultured

Although previous studies have suggested that human peripheral blood mononuclear cells (PBMCs) may express pro-opiomelanocortin (POMC) mRNA and synthesize its related peptides, the patho-physiological role of POMC expressed in peripheral cells is not known. In this study, we investigated the POMC gene expression in various types of human leukemia cell lines by Northern blot analysis and the reverse transcribed-polymerase chain reaction (RT-PCR) method. The POMC mRNA was not detected by Northern blot analysis in all cell lines tested except the Jurkat cell line which is derived from T-lymphoblastic leukemia. The POMC mRNA expressed in the Jurkat cells was smaller than that in the human anterior pituitary gland. The RT-PCR method revealed that a truncated-POMC transcript could be detected not only in lymphoblastic leukemia cells but also in erythroid and myeloid cells. Interestingly, two cell lines of monocytic leukemia, J-111 and U937, did not express the truncated-POMC mRNA. Treatment with concanavalin-A stimulated truncated POMC mRNA expression and ACTH-like immunoreactivity in lymphoblastic leukemia cells with T-(Jurkat) and B-(BALL-1) lymphocyte phenotypes. These results confirm that human leukemia cells except for monocytic cells express a truncated-POMC mRNA as well as in the human normal PBMC.

Topics: Adrenocorticotropic Hormone; Blotting, Northern; Blotting, Southern; Cell Lineage; Concanavalin A; Gene Expression Regulation, Neoplastic; Humans; Jurkat Cells; Leukemia; Leukocytes, Mononuclear; Pro-Opiomelanocortin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured

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

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

Thymosin beta 4 (beta 4) is an ubiquitous 5-kDa peptide that has been identified as an actin-sequestering peptide. In this work, Northern blot analysis was used to study the beta 4 mRNA levels during the cell cycle of rat thymocytes and hepatocytes as well as in human lymphocytes from patients with leukemia. beta 4 mRNA was found in all the stages of thymocyte and hepatocyte cell cycle, showing an increase in the S-phase which was maintained during the G2 and M phases. Incubation of splenic T-cells with concanavalin A, phorbol myristate acetate or the ionophore A23187 lead to a similar increase of beta 4 transcript during the S-phase. The increase in beta 4 mRNA observed in the G2/M boundary of the cell cycle, together with its ability to inhibit actin polymerization, suggests a possible role of beta 4 in the the morphological changes and actin redistribution occurring during the cytokinesis.

Topics: Actins; Animals; Calcimycin; Cell Division; Concanavalin A; Gene Expression; Humans; Leukemia; Liver; Liver Regeneration; Lymphocytes; Peptides; Rats; Rats, Sprague-Dawley; RNA, Messenger; Tetradecanoylphorbol Acetate; Thymosin; Thymus Gland; Time Factors

Carrier-immobilized mono- or disaccharides and other carbohydrate structures, derived by custom-made chemical synthesis, have already proven to be valuable ligands for localizing carbohydrate-binding proteins in tissue sections. Defined purified glycopeptides, as components of neoglycoproteins, offer the possibility of increasing their structural complexity and, thereby, their receptor selectivity. To test the feasibility of this approach, the glycopeptide man6-glcNAc2-asparagine derived from ovalbumin was purified after pronase digestion. It was coupled to bovine serum albumin as carrier protein with the homobifunctional linking agent bis-(sulphosuccinimidyl)suberate to yield the diglycosylated concanavalin A-reactive product. Following biotinylation, it was used to detect mannose-specific binding sites in fixed cells of seven human leukemia or lymphoma lines and in fixed, paraffin-embedded sections of human breast cancer. In comparison to chemically mannosylated bovine serum albumin with ten sites of glycosylation or to ovalbumin, this derivative produced a similar pattern of reaction with a quantitatively lower extent of staining in most cases. Remarkably, the presence of potential endogenous ligands for the detected receptor sites was ascertained using the plant lectin concanavalin A. Thus, the conjugation of a purified, deliberately selected glycopeptide to a suitable carrier produces a histochemical tool for detecting glycopeptide-specific binding sites.

Topics: Asparagine; B-Lymphocytes; Binding Sites; Biotin; Breast Neoplasms; Cell Line; Concanavalin A; Cross-Linking Reagents; Electrophoresis, Polyacrylamide Gel; Histocytochemistry; Humans; Leukemia; Mannose; Oligosaccharides; Pronase; Serum Albumin; Serum Albumin, Bovine; Staining and Labeling; Succinimides; T-Lymphocytes; Tumor Cells, Cultured

The microheterogeneity in bovine serum alpha 1-acid glycoprotein (alpha 1AGP) was examined by crossed affino-immunoelectrophoresis with concanavalin A(Con A). In healthy cows, serum alpha 1AGP revealed 2 fractions which were named fraction 1 and 2 according to the degree of binding affinity to Con A. The mean +/- SD of serum alpha 1AGP concentration was 0.31 +/- 0.09 g/l and the relative amounts of fraction 1 and 2 were approximately 67% and 33% respectively on this method. In bovine leukemia cases, 5 out of 8 had a high serum alpha 1AGP concentration of more than 0.7 g/l. On the electrophoretic pattern, the other 2 fractions (Con A weakly and non-reactive fractions, named fraction 3 and 4) were observed in 5 cases and their relative amounts of 4 fractions were 17.8-28.5%, 35.5-46.0%, 17.9-24.4% and 12.0-17.2%, respectively. In bovie leukemia virus infected cows with no clinical symptoms, however, serum alpha 1AGP concentration and the electrophoretic patterns were almost the same as those in healthy cows.

Topics: Animals; Cattle; Cattle Diseases; Concanavalin A; Female; Immunoelectrophoresis; Leukemia; Leukemia Virus, Bovine; Orosomucoid; Reference Values

BALB/c mice fail to mount significant cell-mediated immunity against the syngeneic virally induced leukemia MCDV-12, and die approximately 10 days after tumor inoculation. Our studies in vitro demonstrated that BALB/c splenocyte and irradiated MCDV-12 cell co-culture led to reduced alloreactivity, including depressed interleukin 2 (IL 2) production. Tumor-induced immune suppression was genetically restricted, antigen nonspecific, and alleviated in part by exogenous IL 2 administration in vitro. Furthermore, mitogen stimulation of IL 2 production, not requiring self or alloantigen recognition, was unaffected by MCDV-12 exposure. These results indicate that tumor cells may escape from immunosurveillance by reducing IL 2 production in the immediate tumor vicinity, and suggest a potential role for major histocompatibility complex antigens in the immunosuppression mechanism.

Topics: Animals; Cell Line; Concanavalin A; Immune Tolerance; Immunity, Innate; Interleukin-2; Leukemia; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred C57BL; Rats; Rauscher Virus; Spleen

Topics: Adolescent; Adult; Concanavalin A; Female; Humans; Leukemia; Male; Middle Aged; T-Lymphocytes, Regulatory

Topics: Child; Concanavalin A; Culture Media; Free Radicals; Hematopoietic Stem Cells; Humans; In Vitro Techniques; Leukemia; Luminescent Measurements; Lymphocytes; Peroxides; Silicon Dioxide

A total of 19 patients, treated for aggressive tumors with high-dose chemo/radiotherapy and autologous bone marrow transplantation (BMT), were studied for concanavalin-A (Con A)-induced proliferation and Con-A-induced cytotoxicity. Ten patients with cytomegalovirus (CMV) antibodies before BMT showed increased Con-A-induced cytotoxicity before and from 100 days after BMT, while Con-A-induced proliferation decreased to less than 10% of control values after BMT and remained so. Nine CMV-negative patients showed normal cytotoxic capacity before and after BMT, while Con-A-induced proliferation recovered slowly from day +30 after BMT. Con-A-induced cytotoxicity was not significantly different between CMV-positive and CMV-negative patients, while Con-A-induced proliferation showed significant differences from day +100 onward.

Topics: Acute Disease; Adolescent; Adult; Antibodies, Viral; Bone Marrow Cells; Bone Marrow Transplantation; Carcinoma; Cell Division; Combined Modality Therapy; Concanavalin A; Cytomegalovirus; Cytotoxicity, Immunologic; Female; Humans; Leukemia; Lymphocyte Activation; Lymphoma; Male; Middle Aged; T-Lymphocytes, Cytotoxic; Testicular Neoplasms

Some characteristics of T cell growth factors derived from adult T cell leukemia virus (ATLV)-transformed cell lines, MT 1 and MT 2 were analyzed. MT 1 cells release significant interleukin 2 (IL 2) activity into the culture medium, which showed the same elution pattern of gel filtration and isoelectric focusing of IL 2 from lectin-stimulated normal human lymphocytes. This activity was also detected in the cell extract of MT 1. In contrast, MT 2 cell line did not produce IL 2 activity, but non-IL 2 type growth factor was observed. The significance of these factors from MT cell lines is discussed from the viewpoint of 'autokine' in ATLV-transformed cells.

Topics: Adult; Animals; Cell Line; Cell Transformation, Viral; Concanavalin A; Deltaretrovirus; DNA Replication; Humans; Interleukin-2; Leukemia; Lymphocyte Activation; Mice; Mice, Inbred Strains; T-Lymphocytes

While the combination of L1210 murine leukemia cell vaccine (L1210 vaccine) with 6-mercaptopurine (6-MP) or 6-thioguanine produces a therapeutic response greater than that induced by either of these agents alone, its combination with cyclophosphamide, N4-behenoyl-1-beta-D-arabinofuranosylcytosine, or 5-fluorouracil does not produce such a response. The administration of cyclophosphamide, N4-behenoyl-1-beta-D-arabinofuranosylcytosine, or 5-fluorouracil alone resulted in a response as great as, or greater than, that induced by 6-MP alone. This and the finding that the 6-MP-induced response was more pronounced upon its delayed rather than its early administration indicate that 6-MP-induced reduction of the tumor burden does not explain this augmentation. The combination of 6-MP and L1210 vaccine was not effective in mice bearing 6-MP-resistant L1210 leukemia; however, an augmented response occurred when the tumor burden was reduced by N4-behenoyl-1-beta-D-arabinofuranosylcytosine, indicating that reduction of the tumor burden by 6-MP was only partially associated with augmentation of the therapeutic response. Augmentation was associated with vaccine-induced antitumor immunity because it was induced by the combination of 6-MP and concanavalin A-bound, but not concanavalin A-free L1210 vaccine. This augmentation was dependent on the timing of the L1210 vaccine administration. The combination was not effective in mice bearing P388 leukemia, indicating the tumor specificity of the augmentation. These results show that 6-MP not only reduced the tumor burden but also potentiated the vaccine-dependent antitumor immunity, resulting in the induction of an augmented therapeutic response.

Topics: Animals; Concanavalin A; Dose-Response Relationship, Drug; Drug Administration Schedule; Drug Synergism; Immunotherapy; Leukemia; Leukemia L1210; Leukemia P388; Leukemia, Experimental; Male; Mercaptopurine; Mice; Thioguanine

We describe a simple, rapid and economical method for the study of the interaction of labeled lectins and the surface membranes of human lymphocytes using a semi-automatic harvesting machine (Titertek Multiple Cell Harvester). The procedure requires both small numbers of cells and small amounts of lectin, moreover it reduces the number of experimental steps required.

Topics: Cell Membrane; Cells, Cultured; Concanavalin A; Cytological Techniques; Humans; Lectins; Leukemia; Lymphocytes; Plant Lectins; Tritium

Rabbit red blood cells have previously been shown to rosette with a subpopulation of thymocytes and with mitogen activated peripheral lymphocytes but not with unstimulated lymphocytes. Using monoclonal antibodies and double marker assays we studied the phenotype of these cells. In thymus, over 90% of rosetting cells express antigens of immature thymocytes (HTA1, OKT6). A proportion of the rosetting cells shows in addition antigens of mature thymocytes (OKT3, UCHT1). These cells probably correspond to a stage of intrathymic maturation between common and mature thymocytes. Virtually all rosetting cells are T cells and express an antigen related to T cell activation (TAC) when lymphocytes are activated by mitogens like PHA or Con A. Few rosetting cells are Ia positive. Two other antigens (OKT9, OKT10) known to be associated with proliferating and immature cells, are found in variable proportions on rosetting cells. After stimulation with allogeneic lymphocytes, fewer rosettes are detected than after stimulation by mitogens. Cells activated by a soluble antigen (PPD) and forming rosettes with rabbit red blood cells have a helper phenotype (Leu3a positive). Screening of leukaemia cell samples revealed that only cells from patients with T-ALL form rosettes with rabbit red blood cells. Rosette formation is almost totally inhibited by a polyclonal anti-thymocyte serum and two monoclonal antibodies (OKT11A,Lyt3) which have been shown to block rosettes with sheep erythrocytes.

Topics: Animals; Antibodies, Monoclonal; Cells, Cultured; Child; Child, Preschool; Concanavalin A; Erythrocytes; Fluorescent Antibody Technique; Humans; Infant; Leukemia; Lymphocyte Activation; Lymphoma; Phenotype; Rabbits; Rosette Formation; T-Lymphocytes

Peripheral T lymphocytes activated in vitro with concanavalin A (Con A) or alloantigens express the thymus leukemia (TL) alloantigen as assessed by staining with the monoclonal antibody TL.m3 and flow cytometric analysis. The determinants detected by TL.m3 on activated cells are encoded within the Tla region and are detected as early as 48 h after activation with Con A. Several long-term cloned cytotoxic T lymphocyte lines were also examined and each expressed TL. By two-dimensional analysis, the TL isolated from activated peripheral cells was indistinguishable from that found on thymocytes and the leukemia cell line ASL-1.

Topics: Animals; Antigen-Antibody Reactions; Antigens, Neoplasm; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Kinetics; Leukemia; Lymphocyte Activation; Membrane Glycoproteins; Mice; Mice, Inbred A; Mice, Inbred BALB C; Mice, Inbred C57BL; Species Specificity; T-Lymphocytes

Eight patients with adult T-cell leukemia (ATL) were observed at Niigata University Hospital during the last seven years. They had typical hematological findings of ATL. All patients tested possessed both natural antibody against ATL-associated antigens (ATLA) in the serum and ATLA in peripheral mononuclear cells. These findings agree with the results of virological analysis in patients with ATL in other areas of Japan and human T-cell leukemia (HTL) in the U.S.A. Six out of the eight patients were native residents of Sado Island in Niigata Prefecture. They showed somewhat different clinical features from patients in other endemic areas and a variety of skin lesions. It was noticeable that the skin lesion regressed spontaneously in two patients. In view of the fact that the total population of this island is about 80,000, the incidence of ATL is thought to be high. Immunological studies were performed in our patients and the immunological background of ATL is discussed.

Topics: Adolescent; Adult; Aged; Antigens, Neoplasm; Concanavalin A; DNA, Neoplasm; Female; Histocompatibility Antigens Class II; HLA-DR Antigens; Humans; Japan; Leukemia; Lymph Nodes; Lymphocyte Activation; Male; Middle Aged; T-Lymphocytes

The content of calmodulin, as measured by a radioimmunoassay, in homogenates of two human leukemic cell lines (IM9 and Molt-4) was about 10-fold higher than that of normal human peripheral lymphocytes. These elevated calmodulin levels existed regardless of the proliferation status of the cells. Normal human lymphocytes stimulated with concanavalin A (con A) did not exhibit these elevated calmodulin levels. Likewise, leukemic cells which were 'blocked' from dividing through the addition of thymidine did not exhibit lower or 'normal' cellular calmodulin levels. These increased levels of calmodulin could contribute to the altered calcium regulation which exists in human leukemic lymphocytes.

Topics: Calcium-Binding Proteins; Calmodulin; Cell Line; Concanavalin A; Humans; Leukemia; Lymphocyte Activation; Lymphocytes; Radioimmunoassay

A simple method is described for preparing monolayers of non-adherent cells, using concanavalin A to bind the cells to wells of plastic microtest plates. The method was used successfully with all 202 human cell types tested, which included 23 tissue culture lines, 165 fresh specimens of all major histological types of leukemia and lymphoma, 20 fresh myelomas, 2 fresh thymomas, normal spleen and lymph node cells, fractionated T lymphocytes and B lymphocytes from peripheral blood, and cultured fetal amniotic cells. All cell types attached firmly, and were not detached by subsequent vigorous washing. In contrast, attempted attachment of cells in serum free medium, or with poly-L-lysine or glutaraldehyde, was ineffective with many cell types. We used the monolayers as target cells for antibodies to cell surface antigens, utilizing immune rosetting or complement-mediated cytotoxicity. This procedure should simplify most assays involving non-adherent target cells.

Topics: Amniotic Fluid; Animals; Antigens, Surface; Cats; Cattle; Cell Adhesion; Cells, Cultured; Concanavalin A; Culture Media; Cytotoxicity, Immunologic; Horses; Humans; Immune Adherence Reaction; Leukemia; Lymphoma; Mice; Rabbits; Rats; Sheep; Swine

Topics: Animals; Cattle; Cattle Diseases; Concanavalin A; Female; Leukemia; Leukemia Virus, Bovine; Leukocytes; Lipopolysaccharides; Mitogens; Phytohemagglutinins; Retroviridae

Lymphocyte activating factor (LAF) is a monocyte derived product which has the ability to enhance the proliferative response of murine thymocytes to the T cell mitogen concanavalin A (Con A). In this report, we have demonstrated that supernatants from normal human polymorphonuclear cells (PMNs) and from granules obtained from PMNs exhibit activity identical to LAF when tested against mouse thymocytes. Our results suggest that PMNs may release factors essential for T cell activation and thus, like monocytes, may serve as important accessory cells in the development of T cell-dependent immunity.

Topics: Animals; Concanavalin A; Humans; Interleukin-1; Leukemia; Lymphocyte Activation; Mice; Neutrophils; Thymidine; Thymus Gland

Topics: Animals; Cell Division; Concanavalin A; Kinetics; Leukemia; Mathematics; Mice; Mice, Inbred CBA; Spleen; Time Factors

The method of cell agglutination with concanavalin 'A' was employed to study lymphocytes from the peripheral blood of cattle spontaneously infected with the virus of leukosis. The infection in the animals was demonstrated through the agar gel immunodiffusion test based on the presence of antibodies. As controls served lymphocytes of normal cattle that were serologically and clinically negative for leukosis as well as lymphocytes, of cattle that were immunized with the virus of the mucous disease and with adeno-, rota, and parainfluenza-3 viruses. It was found that 10(6) lymphocytes of cattle infected with the leukosis virus was agglutinated by concanavalin 'A' used at the rate of 500-2000 microgram/cm3, while the same amount of lymphocytes of normal animals or animals that were immunized against the other viruses got agglutinated by concanavalin 'A' at 4000-8000 g/cm3.

Topics: Agglutination Tests; Animals; Cattle; Cattle Diseases; Concanavalin A; Leukemia; Leukemia Virus, Bovine; Lymphocytes; Retroviridae

Topics: Cell Line; Concanavalin A; Cyclic GMP; Enzyme Inhibitors; Guanylate Cyclase; Humans; Leukemia; Lymphocyte Activation; Lymphocytes; Plant Extracts

Normal and leukaemic lymphoid cells, both human and murine, were stained for specific carbohydrates with three fluorescein-labelled lectins: Aprotinin for sialyl (or uronyl) groups: Ricinus agglutinin for galactosyl groups; and Concanavalin A for mannosyl (or glucosyl) groups. The method gives permanent preparations of sections from methanol fixed, paraffin embedded tissues, from blood and bone marrow films or touch preparations of lymph nodes that were methanol fixed. Whereas normal lymphocytes and lymphoblasts reacted strongly for sialyl groups, lymphoblasts of acute lymphoblastic leukaemia and lymphocytes of chronic lymphocytic leukaemia gave a much weaker reaction. The same was the case of the lymphocytes of the Sézary variant and the lymphocytes of macroglobulinaemia. The fine processes of the cells of hairy cell leukaemia stained well for sialyl groups. No obvious differences were detected between normal monocytes and the cells of monocytic leukaemia, nor between normal plasma cells and those of myeloma.

Topics: Animals; Aprotinin; Carbohydrates; Concanavalin A; Fluoresceins; Humans; In Vitro Techniques; Lectins; Leukemia; Leukemia, Hairy Cell; Leukemia, Lymphoid; Leukemia, Monocytic, Acute; Lymphoid Tissue; Mice; Multiple Myeloma; Waldenstrom Macroglobulinemia

Topics: Aging; Animals; Cell Count; Cell Division; Cell Movement; Concanavalin A; Endotoxins; Leukemia; Lymph Nodes; Lymphocytes; Mice; Mice, Inbred AKR; Mice, Inbred BALB C; Mitogens; Phytohemagglutinins; Spleen; T-Lymphocytes

Topics: 3',5'-Cyclic-GMP Phosphodiesterases; Concanavalin A; Cyclic GMP; Guanylate Cyclase; Humans; Kinetics; Leukemia; Lymphocyte Activation; Lymphocytes; Plant Extracts

Plastic dishes were coated with an agarose layer. The layer was modified by covalently binding proteins to it, using the CNBr-method. Cells were seeded on the dishes and the number of attached cells was evaluated. The specificity of the attachment was demonstrated by showing that cells, carrying specific membrane-bound immunoglobulins, attached only to the corresponding anti-immunoglobulins. This indicated that the method could be used for cell sorting. The attachment of cells to proteins was influenced by the amount of bound protein, incubation time, temperature and the degree of trypsinization. Most attached cells were viable for several days and when dying they detached. Detailed morphological and cytochemical analyses of the dynamics of attachment and cytoplasmic spreading on the chemically well-defined surfaces were possible using the new method.

Topics: Burkitt Lymphoma; Cell Adhesion; Cell Line; Concanavalin A; Glioma; Humans; Leukemia; Lymphoma, Non-Hodgkin; Neuroglia; Polylysine; Polysaccharides; Protamines; Protein Binding; Proteins; Sepharose

Topics: Animals; Cell Membrane; Concanavalin A; DNA, Neoplasm; Humans; In Vitro Techniques; Leukemia; Leukemia L1210; Lymphocytes; Mice

Topics: Agglutination; Animals; Antibody Formation; Cells, Cultured; Concanavalin A; Female; Glutaral; Hot Temperature; Humans; Leukemia; Lymphoma, Non-Hodgkin; Mice; Neoplasm Transplantation; Neoplasms, Experimental; Neuraminidase

Studies of peripheral blood, bone marrow, and spleen cells from three patients with hairy-cell leukemia were performed. Two of the three patients had well-organized cytoplasmic, ribosome-lamellar inclusions in their leukemic cells. Blast transformation and 3H-thymidine incorporation of lymphocytes seemed to fall within normal ranges when the findings were related to the absolute numbers of lymphocytes. The enzymatic markers demonstrated in hairy cells-strong acid phosphatase activity in endoplasmic reticulum and lysosomes, marked alpha-naphthyl acetate esterase reaction, and weak beta-glucuronidase activity-as well as their phagocytosis of latex particles, indicate a common origin with monocytes or histiocytes. No decisive results were obtained by immunofluorescence. Evaluation of the significance of the formation by hairy cells of mouse erythrocyte rosettes, as well as the presence of the typical hair-like projections, may require additional knowledge concerning the membrane of these cells.

Topics: Acid Phosphatase; Adult; Concanavalin A; Esterases; Female; Glucuronidase; Histocytochemistry; Humans; Isoenzymes; Leukemia; Leukocytes; Lymphocyte Activation; Lymphocytes; Male; Microscopy, Electron; Middle Aged; Peroxidases; Phagocytosis

With a newly developed turbidometric method, concanavalin A was shown to agglutinate normal lymphocytes, lymphoma cells, and leukemic cells from chronic lymphocytic leukemia and from acute myelocytic and lymphocytic leukemia. However, there was a marked difference in the kinetics of this agglutination process. Leukemic blast cells and cells from a patient with convoluted lymphoma agglutinated poorly in this system. Conversely, the degree of agglutination for chronic lymphocytic leukemia cells was greater than that for the blast cells and also slightly greater than that for normal lymphocytes. Cultured cells from a Burkitt's lymphoma (Raji) and from a patient with poorly differentiated lymphoma agglutinated very rapidly with concanavalin A. Prior incubation of all cell types with neuraminidase markedly enhanced the agglutination process similar to that of trypsinization. Thus, these studies illustrate the usefulness of this method in quantitating the kinetics of agglutination of various human neoplastic cell types by concanavalin A.

Topics: Agglutination; Concanavalin A; Humans; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Lymphocytes; Lymphoma; Neuraminidase; Trypsin

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

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

Normal peripheral blood leukocytes were tagged with 2, 4-dinitrofluorobenzene at a ratio of 10-11 molecules/cell. One-tenth ml of various concentrations of concanavalin A (Con A) was added to 0.2 ml of either tagged or untagged cells (5 times 10-6/ml) and incubated for 20 minutes at ambient temperature, after which agglutination was scored visually. A readily discernible quantitative difference in the agglutinability of 2, 4-dinitrophenyl (DNP)-tagged versus untagged cells was seen at all concentrations of Con A in the range of 12.5 800 mug/ml. The reaction was maximal at 24 degrees C, somewhat diminished at 37 degrees C, and minimal at 4 degrees C. The agglutination of DNP-tagged leukocytes by 50 mug Con A/ml was completely blocked with 0.1 M methyl-alpha-D-glucopyranoside (alpha-MG), but as low a concentration as 0.001 M alpha-MG inhibited agglutination of untagged cells. The ability of Con A to agglutinate DNP-tagged normal leukocutes may be attributed to a lowering of the zeta potential, a topographic rearrangement of receptor sites, or the formation of new antigenic determinants similar to those found on malignant cells. The last alternative would be consistent with the observation that DNP-tagged normal leukocytes could evoke the production of antibodies that reacted with leukemic granulocytes.

Topics: Agglutination; Concanavalin A; Humans; Leukemia; Leukocytes; Methylglucosides; Nitrobenzenes; Pyrans; Temperature

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: Binding Sites; Bone Marrow; Bone Marrow Cells; Concanavalin A; Fluoresceins; Hodgkin Disease; Humans; Leukemia; Lymphoma; Lymphoma, Non-Hodgkin; Microscopy, Fluorescence; Receptors, Drug

Topics: Animals; Antigens, Neoplasm; Burkitt Lymphoma; Cell Line; Chromatography, Affinity; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Humans; Immune Sera; Iodine Radioisotopes; Leukemia; Leukemia, Myeloid; Mercaptoethanol; Molecular Weight; Papain; Rabbits; Sodium Dodecyl Sulfate; Urea

Topics: Animals; Antibodies, Anti-Idiotypic; Antigen-Antibody Reactions; Azides; Basophils; Binding Sites, Antibody; Cell Membrane; Cells, Cultured; Concanavalin A; Fluorescent Antibody Technique; Immunoglobulin E; Iodine Radioisotopes; Leukemia; Rabbits; Rats; Temperature

Topics: Animals; Antigens, Neoplasm; Cattle; Cells, Cultured; Chromium Isotopes; Concanavalin A; Culture Media; Cyclic AMP; Cytotoxicity Tests, Immunologic; Dactinomycin; Epitopes; Fetus; Humans; Immunity, Cellular; Lectins; Leukemia; Lymphocyte Activation; Lymphocytes; Lymphoid Tissue; Models, Biological; Radiation Effects

Topics: Animals; Antigens, Neoplasm; Concanavalin A; Cytotoxicity Tests, Immunologic; Immunization; Lectins; Leukemia; Leukemia, Experimental; Lymphocytes; Lymphoma, Non-Hodgkin; Mice; Neoplasm Transplantation; Sarcoma, Experimental