concanavalin-a and Burkitt-Lymphoma

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

The expression of cell surface oligosaccharides is associated with various biological phenomena. To clarify the relationship between lectin binding and the survival of patients with Burkitt's lymphoma, tumor samples from nine patients with Burkitt's lymphoma were analyzed by lectin histochemistry. Kaplan-Meier analysis showed that survival is significantly shorter for patients with negative reactivity for lectins from Phaseolus vulgaris (L-PHA), Arachis hypogaea (PNA), or Canavalia ensiformis (ConA) than for those with positive reactivity for these lectins. Immunohistochemistry for N-acetylglucosaminyltransferase V, which synthesizes beta1,6-branched oligosaccharides such as L-PHA-reactive oligosaccharides, was positive in 8 of 9 patients, but there was no correlation between its expression and that of L-PHA-reactive oligosaccharides. Collectively, a loss of L-PHA-, PNA-, or ConA-reactive oligosaccharides is closely associated with a poor prognosis in patients with Burkitt's lymphoma.

Topics: Adolescent; Adult; Burkitt Lymphoma; Child; Child, Preschool; Concanavalin A; Humans; Immunohistochemistry; N-Acetylglucosaminyltransferases; Oligosaccharides; Peanut Agglutinin; Phytohemagglutinins; Prognosis

We investigated the expression of differential lambda light chains in human B cell lines secreting immunoglobulin (Ig). When these cell lines were cultured with concanavalin A for a long period of time, a subpopulation of some but not all of these cell lines was induced to express new lambda light chains replacing the original lambda chain (light chain shifting). Production of the new lambda chain, which replaces the original lambda chain, results from a VJ rearrangement at a previously excluded allele and a dramatic reduction of the original lambda chain transcript, although no difference was found in the level of heavy chain transcript. Recombination activating genes RAG-1 and RAG-2, which are normally expressed during specific early stages of lymphocyte development, were expressed in not only the light chain shifting-inducible lines but also in the non-inducible cells. Treatment of these Ig secreting cell lines with dibutyryl cAMP, which is known to enhance expression of the RAG genes, could not induce the creation of new lambda light chain-producing cells from the inducible lines, suggesting that the expression of the two RAG genes is not sufficient for inducing new lambda light chain production. Concanavalin A induced a gradual but significant production lost of the original lambda chain in a subpopulation of the light chain shifting-inducible cells but not in the non-inducible cells. Association of new lambda light chain production with loss of original lambda chain raises the possibility that, when the RAG genes are expressed, concanavalin A may act on a novel intracellular mechanism controlling lambda light chain allelic exclusion in these plasma cell lines.

Topics: Adenocarcinoma; Alleles; B-Lymphocytes; Base Sequence; Burkitt Lymphoma; Cell Line; Concanavalin A; DNA Primers; DNA-Binding Proteins; Flow Cytometry; Gene Rearrangement; Genes, Immunoglobulin; Homeodomain Proteins; Humans; Hybridomas; Immunoglobulin lambda-Chains; Lung Neoplasms; Molecular Sequence Data; Nuclear Proteins; Polymerase Chain Reaction; Protein Biosynthesis; Transcription, Genetic

Immune responses in lymphocytes require cellular accumulation of large amounts of calcium (Ca2+) from extracellular sources. In the T cell tumor line Jurkat, receptors for the Ca(2+)-releasing messenger inositol 1,4,5-trisphosphate (IP3) were localized to the plasma membrane (PM). Capping of the T cell receptor-CD3 complex, which is associated with signal transduction, was accompanied by capping of IP3 receptors. The IP3 receptor on T cells appears to be responsible for the entry of Ca2+ that initiates proliferative responses.

Topics: Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Burkitt Lymphoma; Calcium; Calcium Channels; CD3 Complex; Cell Line; Cell Membrane; Cells, Cultured; Concanavalin A; Fluorescent Antibody Technique; Humans; Inositol 1,4,5-Trisphosphate; Inositol 1,4,5-Trisphosphate Receptors; Kinetics; Receptors, Antigen, T-Cell; Receptors, Cell Surface; Receptors, Cytoplasmic and Nuclear; Second Messenger Systems; T-Lymphocytes

Two lymphoblastoid tumor cell lines, the Burkitt lymphoma derived BJAB cell line which is free of Epstein-Barr virus (EBV) and B95-8 cells, which are marmoset lymphocytes transformed by EBV isolated from an infectious mononucleosis patient, were studied in regards to their effects on the blastogenic responsiveness of normal human peripheral blood leukocytes stimulated in vitro with mitogens. Mitomycin C treated tumor cell suspensions, when cocultured with normal human blood leukocytes, markedly depressed the expected blastogenic responses in vitro to concanavalin A, pokeweed mitogen, and phytohemagglutin. In addition, cell-free sonicates from the cell lines also depressed blastogenic responsiveness of the leukocytes in vitro. Heating the sonicates for 10 min at 100 degrees C markedly diminished the suppressive properties of the sonicates, as did ultraviolet light irradiation. The suppressive activity of the B95-8 sonicates was pelleted by high speed centrifugation as compared to the activity of sonicates derived from the BJAB cells. Further studies are warranted to determine the nature and mechanism of suppression of blastogenic responsiveness of normal human leukocytes by soluble components derived from such lymphoblastoid cell lines.

Topics: Animals; Burkitt Lymphoma; Callitrichinae; Cell Line; Cell Transformation, Viral; Cell-Free System; Concanavalin A; DNA; Herpesvirus 4, Human; Humans; Lymphocyte Activation; Lymphocytes; Mitogens; Mitomycin; Mitomycins; Phytohemagglutinins; Pokeweed Mitogens

To evaluate the interactions between circulating immune complexes (CIC) and lymphoid cells in primary biliary cirrhosis (PBC), we determined (1) whether antibodies to lymphocytes in PBC serum, independent of CIC, could account for binding in the Raji cell assay for CIC and (2) whether CIC or other humoral factors in PBC serum could interact with lymphoid cells to alter their function. We found that three quarters of CIC positive PBC sera bound specifically to Raji cells via complement receptors, while only one quarter had antibodies to lymphoid cells or Raji cells devoid of complement receptors. We also demonstrated factors which inhibited cell-mediated cytotoxicity and suppressor cell activity in PBC sera; however, we found no correlation between the level and presence of CIC or of lymphocyte antibodies and the level or presence of these serum inhibitory factors. Thus, although the detection of CIC in PBC is not artifactual, the contribution of CIC and other serum factors to the other immunological aberrations in PBC remains to be elucidated.

Topics: Antibodies, Neoplasm; Antigen-Antibody Complex; Antilymphocyte Serum; Burkitt Lymphoma; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Liver Cirrhosis, Biliary; Lymphocytes; T-Lymphocytes, Regulatory

A new technique for the detection of glycoprotein antigens in immune complexes (IC) isolated from serum is described. The technique was developed with a model IC system consisting of ovalbumin (OVA)-rabbit anti-ovalbumin antibodies (aOVA), at 3 times antigen excess. OVA-aOVA IC added to normal human serum (NHS) were purified by absorption onto and elution from tubes coated with rheumatoid factor (RF) and were subjected to electrophoresis in polyacrylamide gels. Concanavalin A (Con A)-binding proteins were detected by treating the gels with radioiodinated Con A (125Con A), followed by autoradiography. IC isolated from sera of patients with Burkitt's lymphoma (BL) and Nasopharyngeal Carcinoma (NPC) were analyzed before and after reduction with dithiothreitol. Two closely spaced proteins of about 40 kdalton were identified in the reduced samples in 26 of 30 BL sera (86%) and in 24 of 30 NPC sera (80%) but were not seen in 30 sera of African patients with a variety of unrelated tumors nor in 12 sera of European blood bank donors.

Topics: Antigen-Antibody Complex; Antigens, Neoplasm; Burkitt Lymphoma; Carcinoma; Concanavalin A; Glycoproteins; Humans; Molecular Weight; Nasopharyngeal Neoplasms; Prognosis

Topics: Animals; Antibiotics, Antineoplastic; Burkitt Lymphoma; Cell Line; Cell Membrane; Concanavalin A; Cytoskeleton; Fibroblasts; Humans; Immunologic Capping; Lymphocytes; Mice; Microtubules; Zinostatin

Topics: Burkitt Lymphoma; Cell Line; Cell Transformation, Neoplastic; Cell Transformation, Viral; Concanavalin A; Herpesvirus 4, Human; Humans; Infectious Mononucleosis; Lectins; Lymphocyte Activation; Lymphocytes; Protein Biosynthesis

Topics: B-Lymphocytes; Burkitt Lymphoma; Cell Differentiation; Cell Line; Cell Transformation, Neoplastic; Concanavalin A; Herpesvirus 4, Human; Humans; Lymphocyte Activation; Membrane Proteins; Molecular Weight; Neoplasm Proteins; T-Lymphocytes

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

Con A-Sepharose affinity chromatography may be used in the analysis and classification of immune complexes. Experiments with model immune complexes suggest that the degree of affinity of an immune complex for Con A-Sepharose is determined by the antigen rather than the IgG antibody of the complex. It is possible that partial characterization of unknown antigens linked to IgG in immune complexes may be achieved in many diseases. Preliminary explorations with selected human sera indicate that the IgG containing immune complexes in Burkitt's lymphoma and nasopharyngeal carcinoma have affinity for Con A-Sepharose. By contrast IgG containing immune complexes in chronic hepatitis B seem to lack affinity for Con A-Sepharose.

Topics: Antigen-Antibody Complex; Burkitt Lymphoma; Chromatography, Affinity; Chronic Disease; Concanavalin A; Hepatitis B; Humans; Immune Sera; Immunoglobulin G; Nasopharyngeal Neoplasms; Sepharose

The isolation and establishment in vitro of a hitherto undescribed type of lymphocyte designated D.G.-75 is reported. The original inoculum was derived from the pleural effusion of a child with a primary abdominal lymphoma, which clinically and histologically resembled Burkitt's lymphoma. In addition to the absence of the EBV genome and EBV receptors, this line possesses a number of other properties which distinguish it from previously described lymphoblastoid cell lines. It has different growth characteristics and morphology; does not form EAC or E rosettes (representative of B and T) cell surface markers, respectively); possesses IgM-kappa immunoglobulins on the cell surface (B lymphocyte), has an unusually high cap-forming ability and low agglutinability with fluorescent concanavalin A. One homologue of the No.14 chromosome pair possesses extra chromatin material as revealed on chromosome banding. This abnormal chromosome marker is similar to that described in biopsies and cultured tumor cells from patients with African Burkitt's lymphoma.

Topics: Agglutination Tests; Antibodies, Viral; Burkitt Lymphoma; Cell Line; Child; Chromosome Aberrations; Chromosomes, Human, 13-15; Concanavalin A; Herpesvirus 4, Human; Humans; Immunoglobulin kappa-Chains; Immunoglobulin M; Lymphocytes; Male; Receptors, Antigen, B-Cell

Topics: Agglutination Tests; B-Lymphocytes; Burkitt Lymphoma; Cell Membrane; Concanavalin A; Hodgkin Disease; Humans; Leukemia, Lymphoid; Lymph Nodes; Lymphocytes; Lymphoma; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Receptors, Concanavalin A; Remission, Spontaneous; Spleen

Sera of individuals with Burkitt's lymphoma and nasopharyngeal carcinoma, tested by consumption of hemolytic complement, were found by comparison with healthy individuals to have significantly increased levels of circulating immune complexes. The identity of the immune complexes was established by sucrose density gradient ultracentrifugation, which showed them to sediment between 10 and 19S. As they were adsorbable by rheumatoid factor--Sepharose 4B conjugates, it appeared that these complexes were composed of IgG. The complexes were retained by Concanavalin A--Sepharose columns and eluted by alpha methyl-D-mannoside, suggesting by analogy with model complexes that the antigens might be glycoproteins.

Topics: Adsorption; Antibodies, Heterophile; Antigen-Antibody Complex; Antigens, Heterophile; Burkitt Lymphoma; Cell Line; Centrifugation, Density Gradient; Chromatography, Affinity; Complement C3; Complement System Proteins; Concanavalin A; Hemolytic Plaque Technique; Humans; Immunoglobulin G; Male; Nasopharyngeal Neoplasms; Rheumatoid Factor; Sepharose

Human peripheral blood lymphocytes stimulated with concanavalin A for 72 hr have a 10-fold greater capacity to repair DNA damage induced by N-acetoxy-2-acetylaminofluorene than do unstimulated cells. The increased capacity of concanavalin A-activated cells to repair DNA is not observed after 24 hr in culture, a time at which stimulated cells have not begun to synthesize DNA. The maximum rate of repair synthesis obtained after treatment of stimulated cells with the "large patch"-inducing agent, N-acetoxy-2-acetylaminofluorene, is twice that obtained with methyl methanesulfonate, an agent inducing "small patch" repair. The difference between the maximum rates obtained with N-acetoxy-2-acetylaminofluorene and methyl methanesulfonate is 6-fold in a human lymphoblastoid line. Unstimulated lymphocytes show almost identical rates of repair after treatment with either N-acetoxy-2-acetylaminofluorene or methyl methanesulfonate. There is close correlation between the rate of N-acetoxy-2-acetylaminofluorene-induced repair synthesis and the loss of acetylaminofluorene adducts from DNA. Treatment of lymphocytes with methyl methanesulfonate leads to degradation of cellular DNA with the production of single-stranded regions. Such degradation is not observed with N-acetoxy-2-acetylaminofluroene. We conclude that the rate of excision repair is a function of the capacity of cells for DNA synthesis and that lymphocytes that do not synthesize DNA have a limited repair capacity and cannot be used to distinguish between large and small patch repair.

Topics: Acetoxyacetylaminofluorene; Burkitt Lymphoma; Carcinogens; Cell Line; Cells, Cultured; Concanavalin A; DNA; DNA Repair; Hydroxyurea; Lymphocytes; Methyl Methanesulfonate; Time Factors

Lymphocytes isolated from the peripheral blood and from tumor tissues of patients with African Burkitt's lymphoma have been studied for cap formation and agglutinability by Concanavalin A (Con A). Peripheral blood from healthy adult persons served as a normal control and blood from patients with carcinoma served as a non-lymphoma control. These studies included 29 patients with Burkitt's lymphoma, 93 with carcinoma, and 105 healthy adult persons, as well as tumor tissues from 13 patients with Burkitt's lymphoma. The great majority of the carcinomas were from the face and neck regions. Lymphocytes from the blood of the majority of patients with Burkitt's lymphoma, as well as those from tumor tissues, exhibited a reduced cap-forming ability (2-6%) and increased Con-A-induced agglutinability compared to lymphocytes from healthy normal donors and from patients with carcinoma, although some of the lymphocytes from patients with carcinoma had a somewhat lower range of cap formation than the lymphocytes from healthy donors. No difference was observed in the interaction with Con A of lymphocytes from the different types of carcinoma studied. Eight lymphoid cell lines were established in our laboratory from the tumor tissues of patients with Burkitt's lymphoma. The cap-forming ability and agglutinability by Con A of these lines was examined and compared to those of the "classical" lymphoma lines: Raji, Daudi and P3HR1. All cell lines exhibited an increased Con-A-induced agglutinability and a reduced cap-forming ability compared to normal lymphocytes, except for P3HR1 cells which exhibited a cap-forming ability of 15-20%. These findings are discussed in relation to the association of the lymphocytes with malignancy and as a possible aid in the differential diagnosis between malignant lymphomas and other diseases.

Topics: Agglutination; Burkitt Lymphoma; Cell Line; Cell Membrane; Concanavalin A; Head and Neck Neoplasms; Humans; Lymphocytes; Lymphoma; Receptors, Drug

Topics: Animals; Burkitt Lymphoma; Cell Line; Concanavalin A; DNA; Lipopolysaccharides; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Macrophages; Peritoneum; Polysaccharides, Bacterial; Rats; Rats, Inbred Lew; Rats, Inbred Strains; Salmonella; Spleen; Thymidine; Thymus Gland; Tritium

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

Topics: Animals; Bacterial Proteins; Burkitt Lymphoma; Cell Membrane; Chondroitin ABC Lyase; Chondroitin Sulfates; Cold Temperature; Concanavalin A; Electrophoresis; Ethylmaleimide; Extracellular Matrix; Glycosaminoglycans; Glycosylation; Humans; Iodoacetamide; Melanoma, Experimental; Mice; N-Acetylneuraminic Acid; Neuraminidase; p-Chloromercuribenzoic Acid; Phytohemagglutinins; Sulfhydryl Reagents; Surface Properties; Tumor Cells, Cultured; Ultraviolet Rays; X-Rays

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: Agglutination Tests; Agglutinins; Burkitt Lymphoma; Cell Line; Concanavalin A; Humans; Lectins; Methods; Mitogens; Mitosis; Stimulation, Chemical; Temperature; Trypsin