concanavalin-a and Leukemia--Lymphoid

Heterogeneous distribution of surface domains is a characteristic feature of the tumor cell surface and the distribution differs from that of normal cells. During the malignant transformation the heterogeneity may change or disappear. Cell lines with various metastasizing capacities show different distributions of membrane domains or other differences in membrane or surface organization. We have demonstrated that the amount and distribution of negatively charged sites of B 16 melanoma membranes changed upon ionizing radiation (X-ray, 60Co-gamma). In the case of the P 388 lymphoma, however, only the amount of negatively charged sites change after irradiation, the distribution remains unaltered. Both features proved to be radioresistant in human lymphoid leukemic cells.

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Ferritins; Histocytochemistry; Humans; Leukemia, Lymphoid; Lymphoma; Male; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Microscopy, Electron; Protein Binding; Tumor Cells, Cultured

Topics: Cell Line; Concanavalin A; Humans; Interleukin-2; Kinetics; Leukemia, Lymphoid; Lymphocyte Activation; Lymphokines; Palatine Tonsil; Phytohemagglutinins; Receptors, Immunologic; T-Lymphocytes

Topics: Antibody Formation; B-Lymphocytes; Clone Cells; Concanavalin A; Dextrans; DNA; Humans; Immunoglobulins; Leukemia, Lymphoid; Lipopolysaccharides; Lymphocyte Activation; Phytohemagglutinins; Pokeweed Mitogens; Tuberculin

Topics: Agammaglobulinemia; Animals; B-Lymphocytes; Cells, Cultured; Concanavalin A; DNA; Fetal Blood; Hemolytic Plaque Technique; Herpesvirus 4, Human; Humans; Immune Sera; Immunoglobulin G; Immunoglobulin M; Infectious Mononucleosis; Leukemia, Lymphoid; Lymphocyte Activation; Pokeweed Mitogens; T-Lymphocytes

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

Expression of two developmentally regulated carbohydrate antigens, the sialyl stage-specific embryonic antigen-1 (SSEA-1) and I-antigens, in human lymphocytes and lymphocytic leukemia cells was investigated using specific monoclonal antibodies. Sialyl SSEA-1 was expressed only on natural killer (NK) cells, and was essentially absent on resting mature T and B cells among normal peripheral lymphocytes. On the other hand, the I-antigen was strongly expressed on virtually all mature B cells, moderately expressed on most mature T cells, but not expressed on NK cells in normal donors. Expression of the two antigens on normal T and B cells was reversible; in vitro stimulation of normal lymphocytes with concanavalin A (Con A) resulted in the loss of I-antigen and appearance of sialyl SSEA-1 on CD3+ T blasts, whereas stimulation with pokeweed mitogen led to loss of I-antigen expression and appearance of sialyl SSEA-1 antigen on CD19+ B blasts. Among lymphocytic leukemia cells, sialyl SSEA-1 was detected primarily on leukemia cells having immature properties such as most common acute lymphocytic leukemia (cALL) blasts, while the I-antigen was frequently expressed on malignant cells having relatively mature properties, such as those found in adult T-cell leukemia or chronic lymphocytic leukemia, and only occasionally on cALL blasts. Among normal peripheral lymphocytes, the sialyl SSEA-1+I-antigen- NK cells selectively underwent E-selectin (ELAM-1, endothelial-leukocyte adhesion molecule-1)-dependent adhesion to endothelial cells, while the I-antigen+sialyl SSEA-1- mature T and B cells did not, in line with the recent finding that sialyl SSEA-1 serves as a specific ligand for E-selectin. Con A blasts, which are sialyl SSEA-1+I-antigen-, also exhibited significant E-selectin-dependent adhesion to endothelial cells. These results indicate that expression of the sialyl SSEA-1 and I-antigens varies alternately depending on the differentiation/activation status of the lymphocytes, and that this at least partly regulates the behavior of lymphocytes at the vessel wall.

Topics: Antibodies, Monoclonal; B-Lymphocytes; Carbohydrate Sequence; Carbohydrates; Cell Adhesion; Cell Differentiation; Cell Line; Concanavalin A; Endothelium, Vascular; Glycosphingolipids; Humans; Killer Cells, Natural; Leukemia, Lymphoid; Lewis X Antigen; Lymphocyte Activation; Lymphocytes; Molecular Sequence Data; Pokeweed Mitogens; T-Lymphocytes

In our experiment, lymphocyte membrane was labeled by DPH fluorescence probe. The rate of rotation of the probe can be measured from the value of fluorescence polarization (PDPH). With this method useful information could be provided about membrane fluidity of lymphocytes. It was found that the F value (unit of lipid fluidity of membrane) of leukemic lymphocytes was obviously higher than that of normal ones. Furthermore, the F value of cultured leukemic Ts lymphocytes was the highest. In contrast with normal spleen T-lymphocytes or mixed lymphocytes, the response of malignant lymphocytes to the stimulation with ConA or PHA was reflected in the decrease of PDPH value or the increase of F value. Unexpectedly, the F value of T-lymphocytes from "615" mouse not injected with tumour cells was also higher than that of the mixed. The possibility of using the membrane fluidity as a diagnostic criterion was also discussed.

Topics: Animals; Concanavalin A; Fluorescence Polarization; Leukemia, Experimental; Leukemia, Lymphoid; Male; Membrane Fluidity; Mice; Phytohemagglutinins; T-Lymphocytes

The effect of large granular lymphocyte leukemia on F344 rat lymphocytes was studied by analyzing the blastogenic responses of normal spleen cells exposed to serum from leukemic rats. Sera from both transplanted as well as spontaneous cases of LGL leukemia markedly depressed the response to ConA and PHA. The suppressive activity was heat-stable at 56 degrees C, non-dialyzable and was effective even when added to lymphocytes only during the last 24 hrs of culture. A similar depression of blastogenesis was caused by sera from nonleukemic fasted rats. Depletion of lipoproteins from sera partially relieved the suppression. Leukemic and fasted rats had nearly identical serum lipoprotein electrophoretic profiles, indicating that abnormal lipoprotein metabolism may result from the severe anorexia which characterizes the terminal stages of the disease and may cause immunosuppression. Residual suppressive activity was also found in lipoprotein depleted sera. Supernatant fluids from spleen cell cultures of some leukemic rats also depressed lymphocyte blastogenic responses when compared to supernatants from normal spleen cultures.

Topics: Animals; Concanavalin A; In Vitro Techniques; Leukemia, Lymphoid; Lipoproteins; Lymphocyte Activation; Male; Phytohemagglutinins; Rats; Rats, Inbred F344; Spleen; Suppressor Factors, Immunologic; T-Lymphocytes

We studied the role of total peripheral blood T-lymphocytes and separated subpopulations of OKT4+ and OKT8+ cells stimulated with concanavalin A in the regulation of human granulopoiesis. Unfractionated total T cells depleted of monocytes are capable of producing colony-stimulating activity (CSA) and colony-forming unit-clone (CFU-C) suppressor activity simultaneously. After positive selection of cell subsets using a fluorescence-activated cell sorter, these two activities were produced by OKT4+ lymphocytes, whereas OKT8+ cells displayed small amounts of CSA and were incapable of releasing suppressor activity. On the other hand, total human thymocytes and subsets defined by monoclonal antibodies Leu2a/Leu3a failed to express any detectable CSA or CFU-C suppressor activity. A total of 14 cases of phenotyped lymphoid malignancies were also studied: the results showed that the production of stimulating and/or inhibiting factors is neither clearly related to a discrete stage of differentiation nor to the OKT4/OKT8 phenotype. Moreover, three monoclonal T leukemias, OKT4+OKT8-, OKT4-OKT8-, and OKT4-OKT8+ have been able in each case to produce simultaneously CSA and CFU-C suppressor activity. Finally, these studies strongly suggest that the activities of T-lymphocytes involved in the regulation of granulopoiesis are not closely linked with OKT4/OKT8 phenotype.

Topics: Cell Adhesion; Cell Communication; Cell Line; Child; Colony-Forming Units Assay; Colony-Stimulating Factors; Concanavalin A; Granulocytes; Hematopoiesis; Hematopoietic Stem Cells; Humans; Killer Cells, Natural; Leukemia, Lymphoid; Lymphocyte Activation; T-Lymphocytes; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Helper-Inducer

Conditioned medium (CM) from 24-hr culture of guinea pig L2C B lymphoblastic leukemia cells contained an inhibitor(s) of mitogen- and antigen-stimulated proliferation of syngeneic (strain 2 guinea pigs), allogeneic (Hartley guinea pigs), and xenogeneic (Balb/c mouse, NZW rabbit) lymphocytes. The proliferation of several lymphoid and nonlymphoid cell lines also was inhibited in the presence of CM. The inhibitor(s) in CM was not toxic to any of the cultures studied. CM inhibited the mitogen-stimulated proliferation of lymphocytes when added to cultures up to 52 hr after addition of mitogen. Normal responsiveness to mitogens could be restored by washing the CM-treated lymphocytes with medium during the first 6 hr of culture. The addition of exogenous IL-2 to lymphocyte cultures did not overcome the CM-mediated suppression of mitogen- or antigen-stimulated proliferation. CM also inhibited the IL-2-dependent proliferation of murine CTLL-2 cells. Preincubation of guinea pig lymphocytes in CM did not inhibit the capacity of these cells to release IL-2 after exposure to mitogen. The antiproliferative activity of CM was stable to heating at low pH (100 degrees C, 10 min, pH 4.0), was resistant to treatment with papain, pronase, DNase, and RNase and did not bind to Con A-Sepharose. Incubation of the L2C cells in indomethacin did not inhibit the release of the inhibitor(s). The inhibitor(s) in CM had an apparent molecular weight of 500-3500 Da as determined by dialysis and ultrafiltration analysis. The inhibitory activity was recovered in the organic phase after extraction with chloroform:methanol and eluted distinct from the thymidine standard after gel filtration on Sephadex-G 25. These data suggest that the inhibitor(s) in CM is a nonspecific, low molecular weight, lipid-like component (not prostaglandin) that exerts its antiproliferative effects subsequent to cell activation. The inhibitor(s) did not appear to suppress other biologic functions associated with activation, such as IL-2 secretion. The inhibitor in CM may be important in promoting tumor survival in vivo by suppressing potential anti-tumor cellular immune responsiveness.

Topics: Animals; B-Lymphocytes; Cell Line; Concanavalin A; Culture Media; Cytotoxicity, Immunologic; Guinea Pigs; Immune Tolerance; Interleukin-2; Leukemia, Lymphoid; Lymph Nodes; Lymphocyte Activation; Lymphokines; Mice; Mice, Inbred BALB C; Molecular Weight

The asparagine-linked oligosaccharide structures of chronic lymphocytic leukemia cells (CLL) were studied by sequential lectin affinity chromatography. Glycopeptides were isolated from a Concanavalin A (ConA)-binding glycoprotein fraction prepared from a soluble CLL extract. This fraction, which contained 1.8% of the proteins present in the soluble extract, greater than 30 polypeptides revealed by SDS-PAGE analysis, and known cell surface antigens such as HLA-DR and p85 glycoprotein, must include a major proportion of the glycoproteins of the CLL cells. Glycopeptides were prepared by digestion with pronase, were separated into four pools (A-D) by Bio-Gel P-6 filtration and were radiolabeled by N-14C-acetylation. Glycopeptide pools C and D (0.45 less than Kd less than 0.77) were 95-100% bound to ConA-Sepharose and 7-12% bound to Lens culinaris (Lens)-Sepharose, but did not interact with any of the other lectins tested, suggesting major amounts of high mannose structures and minor amounts of fucosylated biantennary complex structures with terminal GlcNAc on the Man alpha 1-6 arm. Structures with greater than 3 branches were suggested for pools A and B (0 less than Kd less than 0.44) which were 34-65% unbound to ConA-Sepharose and 37-40% bound to Lens-Sepharose. Analysis of the ConA-unbound glycopeptides on RCA-Agarose before and after acid hydrolysis indicated variable amounts of terminal galactose and sialic acid residues. Major components of pool A were structures with terminal GlcNAc on all branches (22%) and fully sialylated structures (20%). In pool B, 20% of the radioactivity interacted with L-Phaseolus vulgaris agglutinin (PHA)-Agarose and with Ricinus communis (RCA)-Agarose, indicative of a fully galactosylated triantennary structure with branches at C-2 and C-6 of the Man alpha 1-6 arm. Half of the L-PHA-interacting material also bound to Lens-Sepharose, indicative of a core fucose residue. A fully galactosylated biantennary complex structure in pool A (13%) was identified by weak binding to ConA-Sepharose and strong interaction with RCA-Agarose. The presence of the polylactosamine sequence (Gal beta 1-4GlcNAc beta 1-3)n on this structure was suggested by a sialic acid independent interaction with wheat germ agglutinin (WGA)-Agarose. A sialic acid dependent WGA-interaction was observed in the ConA-unbound glycopeptides of pool A (5%). Some of the structures identified in this study may be associated with the malignant CLL phenotype and/or with a di

Topics: Asparagine; B-Lymphocytes; Chemical Phenomena; Chemistry; Chromatography, Affinity; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Glycopeptides; Humans; Leukemia, Lymphoid; Oligosaccharides

Normal B lymphocytes are activated, proliferate, and then differentiate into plasma cells and secrete immunoglobulin (Ig). We have reported that chronic lymphocytic leukemia (CLL) T4 cells help and CLL T8 cells lack suppressor effects on Ig synthesis by normal B cells (Blood 62:767, 1983). We have now explored the earlier phase, proliferation, using B cell colony formation; in semisolid media. B lymphocyte colonies from normal individuals and from patients with CLL were grown in 0.3% agarose overlayed with T cells or T cell subsets and the B cell mitogen staphylococcal protein A. Enriched T cells, OKT4 or OKT8, were obtained either by sheep erythrocyte rosettes or depletion of OKT8 or OKT4 cells by monoclonal antibody or complement, respectively. Twenty thousand B cells from normal subjects yielded 65 +/- 9, 64 +/- 7, and 19 +/- 6 colonies with autologous unfractionated T-, OKT4-, or OKT8-positive cells, respectively. This compared to 29 +/- 11, 81 +/- 11, and 15 +/- 4 colonies from patients with CLL with added autologous unfractionated T-, OKT4-, or OKT8-positive cells. To determine whether the fewer number of colonies in both normal subjects and patients with CLL with OKT8-positive cells was due to suppression or lack of help, the number of OKT4-positive cells was held constant, and OKT8-positive cells were added in increasing numbers. No suppression of colony formation could be demonstrated. Furthermore, the addition of increasing numbers of concanavalin A (Con A)-activated OKT8-positive cells did not suppress colony formation. These results suggest that the CLL T cell subsets behave in a functionally similar manner to normal T cell subsets, namely, (1) that normal and CLL B cell colony growth is helped by OKT4 cells; and (2) in contrast to immunoglobulin secretion by B cells, neither normal nor CLL OKT8 cells, unstimulated or activated by Con A, suppress B cell colony growth.

Topics: Aged; Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; B-Lymphocytes; Concanavalin A; Female; Humans; In Vitro Techniques; Leukemia, Lymphoid; Lymphocyte Activation; Male; Middle Aged; T-Lymphocytes; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Phytohemagglutinin-P (PHA-P) and concanavalin A (Con A) induced an increase in sialyltransferase (S-T) activity of the blood lymphocytes within 24 hours. PHA-P induced the maximal increase in S-T activity in the lymphocytes at 10 micrograms/ml and with Con A at 5 micrograms/ml. PHA-P induced a greater response than that by Con A. An additional effect of these mitogens was not observed. The exposure of cell extracts to these mitogens did not induce the increase in S-T activity. PHA-P or Con A had no effect on the induction of terminal deoxynucleotidyl transferase (TdT) activity, TdT-positive cells or peanut agglutinin (PNA)-positive cells from blood lymphocytes. Therefore, the change of the lymphocytes induced by these mitogens does not indicate a reversion to more immature state of lymphocyte differentiation, but represents a process of the functional differentiation to more activated lymphocytes. These mitogens had only a slight effect on S-T activity of human T-lymphoid leukemia cell lines. The decreased effects of the mitogens on S-T activity of leukemic cells may reflect the change of the membranes due to malignant transformation or the incomplete membrane structure in undifferentiated leukemic cells.

Topics: beta-D-Galactoside alpha 2-6-Sialyltransferase; Cell Line; Concanavalin A; DNA Nucleotidylexotransferase; Humans; In Vitro Techniques; Leukemia, Lymphoid; Lymphocytes; Mitogens; Phytohemagglutinins; Sialyltransferases; Transferases

In vitro resting, short-term mitogen stimulated, and proliferating rat thymocytes as well as established human T and B lymphoblastoid cell lines were compared in their capacity to metabolize glucose and glutamine as energy source. Furthermore, the pathways of glutamine metabolism in these cells were studied. Compared with resting thymocytes, glucose metabolism of proliferating thymocytes was 36-fold increased during the incubation; 92% of the amount of glucose utilized was converted into trioses mainly lactate, whereas resting cells metabolized only 38% to trioses. However, the latter oxidized 19% of glucose to CO2, as opposed to 1.1% by the proliferating cells. Rates of glucose uptake and degradation to products by the malignant T lymphoblastoid cell line (Jurkat) were nearly identical with those observed with proliferating rat thymocytes, whereas the benign B lymphoblastoid cell lines (DHg-B-1 and LV-B-1) showed significantly higher rates of glucose metabolism. All three transformed lymphoblastoid cell lines, however, metabolized glucose almost completely to lactate as did the proliferating rat thymocytes. Lymphocytes are able to utilize glutamine with glutamate, aspartate and ammonia being the major end-products. A complete recovery of glutamine carbon in the products was obtained with all cells. Glutamine utilization by incubated proliferating rat thymocytes was 8-fold increased as compared to the resting cells. Again the human T lymphoblastoid cell line showed the same rates of glutamine uptake and conversion into products as did the proliferating rat thymocytes, whereas both B lymphoblastoid cell lines had about 2.5-fold enhanced rates as compared to the T cell line. The results indicate that during lymphocyte proliferation caused by mitogen stimulation as well as by permanent transformation into lymphoblastoid cell lines glucose metabolism is altered not only quantitatively but also qualitatively by changing from partly aerobic to almost complete anaerobic glucose breakdown. Glutamine has been found to be a suitable energy source for lymphocytes. About 75% of the amount of glutamate derived from glutamine entered into the citric acid cycle via the aspartate aminotransferase, and the remaining 25% via the glutamate dehydrogenase reaction. The changes in metabolic rates observed in proliferating as well as in transformed or leukemic lymphocytes appear to be reliable parameters to characterize the state of lymphocyte activation or to evaluate the effi

Topics: Animals; B-Lymphocytes; Cell Line; Concanavalin A; Energy Metabolism; Glucose; Glutamine; Humans; Leukemia, Lymphoid; Lymphocyte Activation; Mitogens; Rats; T-Lymphocytes; Thymus Gland

Using the fluorescent activity staining procedure, transglutaminases in human monocytes, granulocytes and lymphocytes have been characterized with respect to agarose gel electrophoretic mobility and thrombin dependence. A thrombin dependent transglutaminase was found in concanavalin A stimulated peripheral monocytes. The electrophoretic mobility of this zymogen and of platelet and plasma factor XIII was similar. With stimulated peripheral lymphocytes a similar pattern was observed. However, the potential transglutaminase activity in the lymphocyte factor XIII was lower than that in monocytes. Similar results were obtained when lymphocytes from chronic myeloid and chronic lymphocytic leukaemia patients were examined. Quantitatively, the transglutaminase activity in the leukaemic cells was estimated to be lower than the activity in normal cells. With respect to agarose gel electrophoretic mobility, a similar transglutaminase was found in concanavalin A stimulated human peripheral granulocytes. Although electrophoretically clearly separated from tissues transglutaminase, the granulocyte enzyme did not seem to require thrombin for activation.

Topics: Acyltransferases; Blood Platelets; Cell Line; Concanavalin A; Electrophoresis, Agar Gel; Erythrocytes; Granulocytes; Humans; Leukemia, Lymphoid; Leukemia, Myeloid; Leukocytes; Lymphocyte Activation; Monocytes; Transglutaminases

Peripheral blood mononuclear cells obtained from a patient with prolymphocytic leukemia expressed the surface membrane markers characteristic of resting mature T helper lymphocytes. These cells responded to the T cell mitogens PHA and Con A in a blast transformation assay but not the anti-T cell monoclonal antibody Leu 4 and the B cell mitogen, PWM. The concentration of PHA or Con A eliciting maximum blast transformation was less than that required by normal mononuclear cells. The leukemic cells recognised and responded to allogeneic pooled mononuclear cells in a mixed lymphocyte culture. In addition, although they did not express Ia antigens, they served as effective stimulators in the mixed lymphocyte reaction. Consistent with the helper phenotype, the leukemic cells did not produce suppressor factors, but provided help for normal B-enriched lymphocytes to respond to PWM as assessed by both blast transformation and IgG production. T lymphocyte colonies developed when the leukemic cells were treated with PHA during a 20 h liquid culture prior to being seeded into semisolid agar medium containing either PHA or an IL2-containing lymphokine. There was no growth when untreated cells were seeded directly into IL2-containing agar. Analysis of colony formation indicated that, as with normal resting T lymphocytes, proliferation occurred in two distinct steps; activation in response to PHA and replication in response to IL2-like growth factors. These findings demonstrate that in this case the helper T prolymphocytes have the functional capabilities of normal mature T lymphocytes as predicted from their helper phenotype.

Topics: Aged; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Surface; Concanavalin A; Humans; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Male; Microscopy, Electron; Phenotype; Phytohemagglutinins; Rosette Formation; T-Lymphocytes; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

The culture supernatants of Con A-activated human peripheral blood mononuclear cells (PBM) contained at least two regulatory factors upon B cell proliferation. One was B cell growth factor (BCGF), which activated antigen-stimulated B cells to proliferation and clonal expansion, and the other was its inhibitory factor, arbitrarily named B cell growth inhibitory factor (BIF). This BIF inhibited the effect of BCGF on anti-mu-stimulated B cells or the monoclonal mature B cell line (CLL-T.H.) obtained from the peripheral blood lymphocytes of B cell-type chronic lymphocytic leukemia patients, which were activated only with BCGF and without adding other proliferating stimuli (e.g., anti-mu). BIF activity was detected in the 24 hr culture supernatants of Con A-activated human PBM in FCS containing medium and also in serum-free RPMI 1640 medium. This substance with BIF activity could not be derived from FCS. Con A-induced BIF (m.w. of 80,000 and an isoelectric point of pH 5.4) was analyzed by Sephadex G-200 gel filtration and chromatofocusing. BIF was stable at pH 2.0 and at 56 degrees C for 30 min. Partially purified BIF had no effect on cell viability and almost no interferon activity (less than 1 IU/ml). BIF with high titer had a slight but significant inhibition on TCGF-dependent T cell growth and on PHA or Con A responses, but the extent of these inhibitions was far less than that of BCGF-dependent B cell growth. Absorption of BIF with Con A blasts made its inhibition on T cell growth even less. On the other hand, BIF activity could not be absorbed with Con A blasts but was almost absorbed with large numbers of CLL-T.H. cells. BIF had almost no inhibitory effect on the proliferation of a mouse fibroblast cell line (NIH 3T3), a mouse myeloma cell line (NS-1), human lymphoid cell lines (MOLT-4, HSB-2, and Daudi), or a human myeloid cell line (K-562). BIF-producing cells were estimated to be T cells and were identified as T8+ T cells. On the other hand, Con A-induced BCGF was demonstrated to be produced predominantly by T4+ T cells. These results show that human B cell proliferation is regulated by interaction between T4+ and T8+ cells via soluble factors, namely BCGF and BIF, respectively.

Topics: Animals; B-Lymphocytes; Cell Adhesion; Cell Line; Cell Survival; Cells, Cultured; Concanavalin A; Growth Substances; Humans; Interleukin-4; Kinetics; Leukemia, Lymphoid; Lymphocyte Activation; Lymphokines; Mice; Monocytes

T and B lymphocytes from normal individuals and patients with T and B chronic lymphocytic leukaemia (TCLL and BCLL) were induced to spread on a solid surface in the presence of Con A. The proportion of cell circumference exhibiting lamellar activity was considerably greater in T than in B cells. This difference also applied to T and B cells with a single leading lamellipodium. The different pattern of formation of active cell edges implied that the degree of polarity measured as the ratio between the largest and the shortest diameter (over the nucleus) was significantly greater in B than in T cells. This was also obvious when T and B cells, both with a single leading lamellipodium, were compared. The formation of active cell edges in T lymphocytes was generally accompanied by nuclear flattening, even in polar cells with a single leading lamellipodium. B cells, with the exception of one BCLL case, did not exhibit nuclear flattening. Thus, during the entire course of a contact-induced morphogenetic response, T- and B-cell leukaemias were easily distinguishable on the basis of the following criteria: (i) the proportion of the lymphocyte perimeter showing active cell edges, (ii) the degree of polarity and (iii) nuclear flattening.

Topics: Adult; B-Lymphocytes; Cell Adhesion; Cell Membrane; Cell Nucleus; Concanavalin A; Humans; Leukemia, Lymphoid; Microscopy, Electron, Scanning; T-Lymphocytes

An active supernatant (Ly Con A) was prepared by stimulating the normal human lymph node lymphocytes with Concanavalin A (Con A). Peripheral blood lymphocytes (PBL) from 3 healthy subjects and from 8 patients with chronic lymphocytic leukaemia (CLL) were cultured in the presence of Con A and Ly Con A. The latter induced a significant DNA synthesis both in normal and CLL lymphocytes. A proliferative response was still present in CLL after T lymphocytes depletion. The Con A and Ly Con A treatment also induced morphological changes in CLL lymphocytes consistent with plasma cell differentiation. In 3 cases the appearance of cytoplasmic light chains was detected by immunofluorescence. These findings suggest that peripheral blood B lymphocytes from CLL patients can be stimulated to maturation when helper factor(s) released by mitogen activated lymph node lymphocytes are provided.

Topics: Antibody-Producing Cells; B-Lymphocytes; Cell Transformation, Neoplastic; Concanavalin A; Humans; Immunoglobulin Light Chains; Leukemia, Lymphoid; Lymph Nodes; Lymphocyte Activation; Lymphokines; Thymidine

Capping with concanavalin A (ConA) and monoclonal anti-HLA-ABC backbone was studied in childhood acute lymphoblastic leukemia (ALL). Capping with ConA and HLA gave quite different results, both in common ALL and T-ALL. With ConA most cases capped poorly, comparable to results described in chronic lymphatic leukemia and lymphoma, but in several cases capping was comparable to that of normal lymphocytes. In HLA capping T-ALL cells capped better than common ALL cells. HLA capping of T-ALL cells is comparable to that of normal lymphocytes. HLA capping results in handmirror cell formation giving support to the hypothesis that capping and motility are associated events.

Topics: Antibodies, Monoclonal; Bone Marrow Cells; Child; Concanavalin A; HLA Antigens; Humans; Immunologic Capping; Leukemia, Lymphoid; Lymphocytes; Phenotype

The human T lymphoblastoid cell line designated CCRF-CEM responds to phytohemagglutinin with a 3.7-fold enhancement of the 32PO4 incorporation into phosphatidylinositol. In myo-[2-3H]inositol-prelabeled CCRF-CEM cells, phytohemagglutinin induced a 3.3-fold accumulation of myo-[2-3H]inositol phosphate during 15 min incubation at 37 degrees C in the presence of 5 mM LiCl. Since Li+ is a potent inhibitor of myo-inositol-1-phosphatase, the results indicate that phytohemagglutinin induces the hydrolysis of inositol lipids in CCRF-CEM cells. In 32PO4-prelabeled CCRF-CEM cells, phytohemagglutinin induced a breakdown of 28% of [32P]phosphatidylinositol 4,5-bisphosphate 40-60 s after the stimulation. The decrease of [32P]phosphatidylinositol 4,5-bisphosphate was found as early as 10 s after the stimulation. This decrease was followed by an increased 32P-labeling of phosphatidic acid. In [2-3H]glycerol-prelabeled CCRF-CEM cells, phytohemagglutinin induced a transient accumulation of [3H]phosphatidic acid and [3H]diacylglycerol. The amount of [3H]phosphatidic acid in the stimulated cells was 3.7-times the control value at 2 min after the stimulation, whereas the amount of [3H]diacylglycerol in the stimulated cells was 1.5-times the control value at 5 min after the stimulation. In [3H8]arachidonate-prelabeled CCRF-CEM cells, phytohemagglutinin induced a transient accumulation of [3H]phosphatidic acid; the amount was 2.5-times the control value at 2 min after the stimulation. Quinacrine (1 mM) caused 41% reduction in the amount of [3H]phosphatidic acid accumulated by the stimulation in [2-3H]glycerol-prelabeled cells. Stimulation in a Ca2+-free saline containing 1 mM EGTA caused 53% reduction in the amount of [3H]phosphatidic acid accumulated by the stimulation. The results presented in this paper indicate that a human T lymphoblastoid cell line, CCRF-CEM, responds to phytohemagglutinin with a rapid turnover of inositol lipids.

Topics: Cell Line; Concanavalin A; Diglycerides; Glycerides; Humans; Kinetics; Leukemia, Lymphoid; Phosphatidic Acids; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phospholipids; Phosphorus Radioisotopes; Phytohemagglutinins; T-Lymphocytes

In a patient with chronic lymphocytic leukemia (CLL), we previously demonstrated by glucose-6-phosphate isoenzyme analysis that monoclonality was restricted to B lymphocytes. Isolated T cells expressed both isoenzymes and, therefore, were apparently not involved in the leukemic process. This report presents results of a functional analysis of the patient's T cells. Cutaneous anergy to a battery of skin tests was correlated with abnormal proliferative responses of isolated T cells to phytohemagglutinin and concanavalin A. The addition of sufficient numbers of autologous adherent cells restored mitogen responses to nearly normal levels. However, the patient's T cells failed to provide help for differentiation of allogeneic B cells in response to pokeweed mitogen. These data suggest that altered cellular immunity in CLL is not necessarily due to intrinsic T cell abnormalities. Reduced numbers of circulating accessory cells and/or an imbalance in T-cell subsets related to the expansion of the leukemic B-cell clone may play a significant role.

Topics: B-Lymphocytes; Concanavalin A; Female; Humans; Immunity, Cellular; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocyte Cooperation; Middle Aged; Mitogens; Phytohemagglutinins; T-Lymphocytes

The immunoregulatory activity of bone marrow leukemic blasts from five patients with null-cell acute lymphoblastic leukemia (ALL) and two children with T-cell ALL was evaluated. Blasts were studied in co-culture for their effects on the proliferative responses of normal allogeneic peripheral blood lymphocytes to phytohemagglutinin. Mitomycin C-treated null cell ALL blasts from two of five children suppressed the proliferative responses of normal responder cells by 80% and 58% whereas those from two other children enhanced the responses by 118% and 31%. T-cell leukemic blasts from one patient with T-cell ALL exhibited helper cell activity (26%) and T-leukemic blasts from the other demonstrated suppressor cell activity (53%). The helper cell activity of leukemic blasts was associated with stimulating capacity of null blasts in one-way mixed lymphocyte reactions. In six of seven cases, incubation of blast cells with concanavalin A (20 microgram/ml) for 48 hours prior to co-culture with responder cells did not result in the generation of significant suppressor cell activity. Our study suggests that leukemic blasts from certain patients with 'null' and T-cell ALL may possess spontaneous helper or suppressor cell immunoregulatory activity. This functional heterogeneity of leukemic blasts may help in subclassifying the ALL.

Topics: Adolescent; Bone Marrow; Cell Division; Cells, Cultured; Child; Child, Preschool; Concanavalin A; Female; Humans; Leukemia, Lymphoid; Lymphocytes, Null; Male; Mitogens; Mitomycin; Mitomycins; Phytohemagglutinins; T-Lymphocytes; T-Lymphocytes, Regulatory

Fifteen patients with chronic lymphocytic leukaemia (CLL) were studied. The diagnosis was made by a combination of clinical findings, peripheral blood morphology and cell surface markers. The Rai clinical staging (Rai et al, 1975) was used to classify all patients. Serum immunoglobins, B and T cell lymphocytes, TG and TM cell populations were evaluated in all patients before commencing treatment. No correlation was found between clinical staging and the presence or absence of hypogammaglobulinaemia. The ratios of TG to TM cells were abnormal in all patients and this abnormality paralleled the stage of the disease. While there was significant difference in the proliferative responses of highly purified T cells from patients and controls at 1 micrograms and 2 micrograms/ml of Concanavalin A (Con A) (P values = P less than 0.005 and P less than 0.01 respectively). There was no significant difference at dose 5 micrograms/ml. The ability of T cells to suppress allogeneic PBMC responses to Con A was dependent on T cell purification procedures. The significance of the results and the possible role of T cells in the pathogenesis of CLL is discussed.

Topics: Cell Division; Cells, Cultured; Concanavalin A; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Leukemia, Lymphoid; Receptors, Immunologic; T-Lymphocytes; T-Lymphocytes, Regulatory

Quiescent thymocytes, mitogen-stimulated thymocytes and acute-leukaemic lymphoblasts provide a model for the study of protein glycosylation in quiescent cells, mitotically active non-malignant and malignant cells respectively. The biosynthesis of both complex and high-mannose-type oligosaccharides was monitored by metabolic labelling with [6-3]fucose and [2-3H]mannose. Bio-Gel P6 elution profiles of [6-3H]fucose-labelled glycopeptides showed that quiescent thymocytes and stimulated thymocytes synthesized qualitatively and quantitatively similar glycopeptides; however, higher-molecular-weight glycopeptides were synthesized by the acute-leukaemic lymphoblasts. The amount of [2(-3)H]mannose incorporated into glycopeptide by quiescent thymocytes was less than 10% of that incorporated by stimulated thymocytes. The Bio-Gel P6 elution profile of [2(-3)H]mannose-labelled glycopeptides from acute leukaemic lymphoblasts was qualitatively similar to that of stimulated thymocytes, with about 40% of the radioactivity incorporated into one glycopeptide peak. This glycopeptide was characterized by Bio-Gel P6 and concanavalin A affinity chromatography, radioactive-sugar analysis, sensitivity to alpha-mannosidase and endoglycosidase H and resistance to beta-glucosaminidase as containing a high-mannose oligosaccharide, possible of Man7-8GlcNAc2 structure. Pulse/chase experiments indicated that this high-mannose oligosaccharide was an end product and not a biosynthetic intermediate. It is concluded that higher-molecular-weight fucose-labelled glycopeptides are characteristic of the malignant cell type, and the synthesis of high-mannose oligosaccharide, Man7-8GlcNAc2, in stimulated thymocytes and acute-leukaemic lymphoblasts is associated with mitotically active cells.

Topics: Animals; Chromatography, Gel; Concanavalin A; Fucose; Glycoproteins; Leukemia, Experimental; Leukemia, Lymphoid; Lymphocyte Activation; Mannose; Mice; Mice, Inbred AKR; Oligosaccharides; T-Lymphocytes; Tritium

Heteroantisera raised to the acute lymphocytic T (ALL) cell line HSB2 and to Sézary cells react with distinct subpopulations of T lymphocytes. Each antiserum reacts with a different T cell antigen and defines a distinct subpopulation that represents approximately 50% of peripheral blood T lymphocytes. The anti-HSB2-positive subpopulation contained suppressor cells for pokeweed mitogen-dependent immunoglobin (Ig) synthesis whereas the anti-Sézary cell serum-positive population included helper cells for Ig synthesis and mixed lymphocyte responder cells.

Topics: Antigens; Cell Line; Concanavalin A; Cytotoxicity, Immunologic; Humans; Immunoglobulins; Leukemia, Lymphoid; Lymphocyte Culture Test, Mixed; Sezary Syndrome; T-Lymphocytes

T and B lymphocyte function as assessed by pokeweed mitogen stimulation and concanavalin A suppression was studied in 40 children with acute lymphoblastic leukaemia who were in remission and off all treatment. Both plasma cell and immunoglobulin production was reduced in off-treatment patients, particularly in those who had received cyclophosphamide as part of their treatment. The ability to suppress immunoglobulin production was also significantly reduced in these patients. These abnormalities were long-standing and were apparent up to 5 years after stopping treatment with no indication of a return to normality. No clinical correlations have been observed.

Topics: Adolescent; Adult; B-Lymphocytes; Child; Child, Preschool; Concanavalin A; Humans; Immunoglobulins; Leukemia, Lymphoid; Lymphocyte Activation; Pokeweed Mitogens; T-Lymphocytes

Measurements of the intensity and polarization distributions of fluorescein fluorescence in human lymphocyte populations show changes within minutes after exposure of the cells to the mitogens phytohemagglutinin and Concanavalin A. The distributions of polarization, which before mitogen exposure show essentially a single peak, after exposure develop a second peak at a lower value of polarization. Simultaneously, the intensity distributions show a shift to higher levels of intensity. These shifts can be modeled on the basis of a subpopulation of lymphocytes responding to mitogen. Interpretation of the results is complicated by many factors that may influence the shape of the polarization distributions. In particular we show that the measurements are sensitive to the concentrations of phytohemagglutinin, Concanavalin A, Ca++, K+, fluorescein diacetate, the incubation time and the specific donor. Nevertheless, fluorescein fluorescence measurements provide a rapid and sensitive method for studying the physiology of lymphocytes and for identifying responding lymphocyte subpopulations.

Topics: Calcium; Concanavalin A; Cytological Techniques; Dose-Response Relationship, Drug; Fluorescence Polarization; Humans; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Mitogens; Phytohemagglutinins; Potassium

Chronic lymphocytic leukemia (CLL) is, in the great majority of cases, a neoplastic proliferation of B cells. The associated immunologic dysfunction accounts for many of the clinically associated phenomenon such as infection and second malignancies. Much of this dysfunction has been attributed to poor B-cell function. Functional impairment of the T-cell component is currently being elucidated. In the investigation of B cells. The associated immunologic dysfunction accounts for many of the clinically associated phenomenon such as infection and second malignancies. Much of this dysfunction has been attributed to poor B-cell function. Functional impairment of the T-cell component is currently being elucidated. In the investigation of B cells. The associated immunologic dysfunction accounts for many of the clinically associated phenomenon such as infection and second malignancies. Much of this dysfunction has been attributed to poor B-cell function. Functional impairment of the T-cell component is currently being elucidated. In the investigation of B cells. The associated immunologic dysfunction accounts for many of the clinically associated phenomenon such as infection and second malignancies. Much of this dysfunction has been attributed to poor B-cell function. Functional impairment of the T-cell component is currently being elucidated. In the investigation of B cells. The associated immunologic dysfunction accounts for many of the clinically associated phenomenon such as infection and second malignancies. Much of this dysfunction has been attributed to poor B-cell function. Functional impairment of the T-cell component is currently being elucidated. In the investigations reported herein, we further demonstrate the immune deficiency of CLL by measuring proliferation and immunoglobulin synthesis in response to mitogens. Stimulation of various cell fractions indicates that in certain cases there is deficient T-helper, while others T function appears intact. In 2 of 4 cases tested there was augmentation of ig synthesis when the CLL cells were cocultured with thymic epithelium. The implications of such in vitro modulation are discussed.

Topics: Aged; Animals; Cell Separation; Concanavalin A; Female; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Leukemia, Lymphoid; Male; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; Rosette Formation; Sheep; Thymus Gland

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

The function of Con A stimulation was investigated in patients with ALL in remission and in patients with ALL in the acute phase of the disease as compared to healthy controls. Suppression of the Con A response brought about by autologous Con A-activated cells was significantly lower in ALL patients in remission (mean value 29.33%) and in the acute phase (mean value 10%) than in the controls (mean value 64.86%). Suppression of the Con A stimulation of control lymphocytes by Con A-activated homologous cells of ALL patients had a mean value of 51.65%. This was significantly higher than the suppression obtained by the same ALL cells in the autologous system and of the same order of magnitude as the suppression obtained by control Con A-activated cells on control Con A stimulation (57.67%). Suppression of the Con a response of ALL lymphocytes produced by control Con A-activated cells was 23.17% and comparable to healthy controls. These results demonstrate that the function of Con A-induced suppression is significantly lower in ALL patients. They further indicate that at least two cell types are involved in this kind of suppression in humans.

Topics: Acute Disease; Adolescent; Child; Child, Preschool; Concanavalin A; Female; Humans; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Male; T-Lymphocytes, Regulatory

Topics: B-Lymphocytes; Cell Division; Cell Separation; Concanavalin A; Humans; Immunoglobulins; Leukemia, Lymphoid; Lymphocyte Cooperation; Phytohemagglutinins; Receptors, Antigen, B-Cell; T-Lymphocytes

By means of the incorporation rate of 3H thymidine into the lymphocytes of patients with chronic lymphatic leukaemia the possibility of stimulating them by using different mitogens was checked and compared with normal persons. The examination covered 11 patients treated with extracorporeal irradiation of the blood (ECIB), 5 patients treated with a chlorambucil therapy, and 10 untreated patients who were classified according to the staging system proposed by RAI. The lymphocytes of the peripheral blood were stimulated as mixed and isolated T and B-lymphocytes in the microculture by using the mitogens PHA, PWM, ConA, and LPS. In all CLL patients there was a diminished stimulation rate of a mixed lymphocyte population. A relation existed between the seriousness of the stage and the diminution of the incorporation rate of 3H thymidine. A corresponding correlation could not be identified in untreated CLL patients. Isolated T-lymphocytes revealed better results of stimulation than the total population. As to their function B-lymphocytes showed a dependance on the kind of therapy. In the mixed lymphocyte culture of normal persons the best findings could be observed after stimulation with PHA, that is also valid for CLL patients. PHA, PWA, ConA, and LPS were suitable as substances stimulating B-lymphocytes with different efficacy in normal persons and CLL patients. Both collectives showed the best results in the T-lymphocyte culture after stimulation with LPS.

Topics: B-Lymphocytes; Concanavalin A; Humans; Leukemia, Lymphoid; Lipopolysaccharides; Lymphocyte Activation; Mitogens; Neoplasm Staging; Phytohemagglutinins; T-Lymphocytes

Topics: Animals; Concanavalin A; Cytotoxicity, Immunologic; Dose-Response Relationship, Immunologic; Fibroblasts; Humans; Immunoglobulin Fc Fragments; Interferons; Killer Cells, Natural; Kinetics; Leukemia, Experimental; Leukemia, Lymphoid; Leukemia, Myeloid; Lymphocytes

Cell-associated and serum beta2-microglobulin was estimated in seven patients with chronic lymphocytic leukaemia. The amount of cell-associated beta2-microglobulin was significantly reduced (P less than 0.01), due to a decrease in the fraction of beta2-microglobulin that passes unretarded through a concanavalin A affinity column (presumably non-HLA-associated beta2-microglobulin). Serum concentrations of beta2-microglobulin were increased, but no correlation was found between the decrease in cell-associated beta2-microglobulin and the increase in serum beta2-microglobulin. All of the beta2-microglobulin from leukaemic serum was eluted corresponding to a molecular weight of 11,800 and none of it was retarded on a concanavalin A affinity column. The decrease in cell-associated beta2-microglobulin might reflect a change in the qualitative or quantitative expression of beta2-microglobulin-associated membrane structures on the leukaemic cells, perhaps conferring resistance to the cells against hypothetical immunological host defence mechanisms.

Topics: beta 2-Microglobulin; Beta-Globulins; Cell Membrane; Concanavalin A; Humans; Leukemia, Lymphoid; Lymphocytes

Topics: B-Lymphocytes; Concanavalin A; Humans; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Monocytes; Phytohemagglutinins; T-Lymphocytes

Cell surface modifications after vibrio cholerae neuraminidase treatment were investigated using three different tritiated lectins: Concanavalin A, Ricinus sanguineus agglutinin (R.S.A.) and Robinia pseudoacacia lectin. Lectin binding measurements were performed on untreated and enzyme treated cells. The cells used were from chronic and acute leukemic donors. After neuraminidase treatment, a significant increase in the number of receptor sites, from 1 to 3 times, was found in all cases tested and for all three lectins utilized with only one exception. The affinity constant was generally decreased after neuraminidase treatment. The increase in number lectin binding sites, indicating extensive modification of the cell surface, is completely consistant with the known importance of sialic acid in determining immunogenicity.

Topics: Binding Sites; Cell Membrane; Concanavalin A; Humans; Lectins; Leukemia, Lymphoid; Leukocytes; Neuraminidase; Plant Lectins; Plants, Toxic; Receptors, Drug; Ricinus; Sialic Acids

Forty-eight children with untreated acute lymphocytic leukemia were evaluated with regard to their clinical presentation and the surface membrane characteristics, mitogen responsiveness, and cytochemical staining features of their lymphoblasts. The presence at diagnosis of 20% or more bone marrow lymphoblasts possessing T-cell surface markers was associated with the development of early relapse and death. Age, sex, initial peripheral leukocyte count (WBC), lymph node enlargement or the presence of hepato- or splenomegaly bore no statistically significant relationship to the patient's lymphoblast type, and only a WBC in excess of 10(5)/microliter correlated with a poor prognosis. Leukemic T-cells were found to be periodic acid-Schiff negative from nearly all patients, helping to distinguish such patients from the larger group of children whose leukemic cells were PAS positive, but negative for T-cell membrane features. In those patients who had multiple relapses, the surface membrane characteristics, staining features, and mitogen responses of their lymphoblasts remained constant. This suggests that relapse occurs in the same clone of malignant cells present at diagnosis and that the above features may be reliably used to evaluate and classify patients beyond the time of diagnosis.

Topics: Adolescent; Cell Membrane; Child; Child, Preschool; Concanavalin A; Humans; Infant; Lectins; Leukemia, Lymphoid; Leukocyte Count; Lymphocyte Activation; Prognosis; Rosette Formation; T-Lymphocytes

Topics: Aged; B-Lymphocytes; Binding Sites; Cell Membrane; Concanavalin A; Glucosyltransferases; Humans; Lectins; Leukemia, Lymphoid; Lymphocytes; Middle Aged; Phytohemagglutinins; Plant Lectins; Plants, Toxic; Ricinus; Sialoglycoproteins; Triticum

Chronic lymphocytic leukaemia (CLL) has a variable clinical course and there is a great need of new prognostic laboratory parameters in this disease. 24 CLL patients were subjected to routine haematological and clinical investigation. The leukaemic cells were analyzed for surface immunoglobulins, complement and sheep red blood cell receptors, Concanavalin A-induced agglutination, cytoplasmic glucocorticoid receptor and for proliferative activity by measurement of tritiated thymidine incorporation. The surface markers studied indicated that all cases were of B-cell origin. Only 5 of 23 patients studied had normal serum immunoglobulin levels. These cases showed a nonprogressive disease. 8 patients had increased infection tendency, all of whom had subnormal IgG levels; 4 of them also had subnormal IgA and IgM and 2 had subnormal IgA levels. 5 out of 6 patients with progressive disease and 3 of 11 with nonprogressive disease had an increased proliferative index, indicating a correlation between this parameter and disease progression. ConA agglutinability was not correlated to disease activity. Cells from 17 of 22 patients showed measurable amounts of glucocorticoid receptor. The 5 patients lacking this receptor had inactive disease.

Topics: Aged; B-Lymphocytes; Complement C3; Concanavalin A; Dexamethasone; Female; Hemagglutination; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Leukemia, Lymphoid; Male; Middle Aged; Receptors, Antigen, B-Cell; Receptors, Glucocorticoid; Receptors, Steroid; T-Lymphocytes

Sixty-two explants from peripheral blood, bone marrow and cerebral fluid of children with acute lymphoblastic leukemia (ALL) and leukemic transformed non-Hodgkin lymphoma (NHL) were cultivated for at least 8 weeks. Although lymphatic cells persisted up to 16 weeks in tissue culture, no proliferation was observed in 54 cultures. From the remaining cultures, eight permanently growing cell lines were obtained. Five of these were EBNA (Epstein-Barr virus-specific nuclear antigen)-positive. Three, however, were ENBA-negative and lacked Epstein-Barr virus genomes. Two cell lines (KM-3 and SH-2) expressed neither B nor T cell characteristics. One line (JM) expressed T cell characteristics and complement receptors. The growing lymphatic cells represented leukemic cells, since the pattern of cytochemical staining and that of membrane receptors of lymphoblasts from the same donor prior to cultivation were identical. All leukemic cell lines were derived from patients in relapse. In contrast, no proliferation of leukemic cells occurred in explains from patients revealing the first manifestation of the disease. These results suggest enhanced growth potential of lymphoblasts resisting antileukemic therapy.

Topics: Adolescent; Antigens, Viral; beta 2-Microglobulin; Binding Sites; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Child; Complement System Proteins; Concanavalin A; DNA, Viral; Fluorescent Antibody Technique; Herpesvirus 4, Human; Humans; Leukemia, Lymphoid; Lymphocytes; Lymphoma; Male; Microscopy, Electron; Receptors, Antigen, B-Cell; Staining and Labeling; T-Lymphocytes

The purified lymphocytosis promoting factor (LPF) from Bordetella pertussis was found to be a potent mitogen for peripheral blood lymphocytes (PBL) from normal adults as well as for cord blood lymphocytes. Proliferation occurred in autologous plasma or fetal calf serum, regardless of previous exposure to pertussis infection or immunization. Only one adult human serum, from a physician constantly working with B. pertussis, inhibited the mitogenic response to LPF and this serum was shown to contain precipitating antibody against LPF. The proliferative effect of LPF was characteristic of a "nonspecific" mitogen and not of antigen stimulation of sensitized cells.LPF, phytohemagglutinin, and concanavalin A were approximately equal in potency although variation occurred depending upon the cell donor. Experiments with lymphocyte subpopulations obtained by rosetting techniques employing sheep erythrocytes, mouse erythrocytes, and sheep erythrocytes coated with antibody and complement suggested the requirement of a multicellular system for LPF mitogencity.PBL from most patients with chronic lymphatic leukemia and lymphosarcoma cell leukemia were even less responsive to LPF than to phytohemagglutinin, whereas PBL from patients with lymphosarcoma usually responded to both mitogens. It can be inferred from the results of experiments with both normal and leukemic cells that LPF, which is a murine thymus-derived (T)-cell mitogen, is also a T-cell mitogen for human PBL. The exact cell requirement and mode of action, however, are as yet unknown.

Topics: Adult; Bordetella pertussis; Concanavalin A; Humans; Infant, Newborn; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphoma, Non-Hodgkin; Mitogens

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

Topics: Bone Marrow; Bone Marrow Cells; Child; Concanavalin A; Cytological Techniques; Horseradish Peroxidase; Humans; Leukemia, Lymphoid; Leukemia, Myeloid; Leukemia, Myeloid, Acute

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

The response of CLL (chronic lymphatic leukemia) lymphocytes to PHA, PWM, and Con A with respect to changes in surface markers and synthesis and secretion of immunoglobulin were examined. After PHA stimulation the percentage of cells bearing readily detectable surface immunoglobulin (SmIg) diminished rapidly whereas cells forming rosettes with sheep erythrocytes (E-rosettes) increased from less than 1% to 30 to 50%. The great majority of blast-transformed cells were E-rosette-positive cells with a small population of SmIg-positive blast cells also observed. Ig production in four of seven CLL lymphocyte populations was increased 2.5 to greater than 40-fold after 4 to 6 days of culture in the presence of PHA. In contrast, pokeweed mitogen did not affect Ig synthesis. Furthermore, the Ig secreted into the culture supernatant fluids from seven of eight CLL cases examined consisted almost entirely of free light chain molecules. In contrast, the cell lysates contained a significant proportion of intact Ig molecules. These results indicate that CLL cells can, under certain circumstances, be stimulated to synthesize and secrete increased amounts of Ig, but that there is a basic defect in the biosynthetic mechanism of these cells which result in the secretion of free light chains rather than intact immunoglobulin molecules.

Topics: B-Lymphocytes; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Humans; Immunoglobulin Light Chains; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Mitogens; Receptors, Antigen, B-Cell

Mice inoculated with Friend leukemia complex (FLC) pretreated with concanavalin A are resistant to FLC challenge only when they have become infected with the FLC-associated lymphatic leukemia virus (LLV). In interpreting states of resistance to FLC induced by various immunizing procedures, the possibility that immunity is sustained by an unrecognized LLV infection should always be considered.

Topics: Animals; Concanavalin A; Friend murine leukemia virus; Immunization; Leukemia, Experimental; Leukemia, Lymphoid; Mice; Organ Size; Spleen

gamma-Glutamyl transpeptidase, an enzyme that catalyzes gamma-glutamyl transfer from gamma-glutamyl compounds to amino acid and peptide acceptors, and which is known to be localized in the membranes of many epithelial cells, was found in a variety of lymphoid cells. The lymphoid cell enzyme is located on the cell surface, and exhibits substantially the same substrate specificity as the enzyme found in epithelial cells. Human and rat (but not mouse) lymphocyte gamma-glutamyl transpeptidase was stimulated by treatment of the cells with mitogens. Normal human peripheral B-cells were more active than T-cells, but the reverse relationship of activities was found in chronic lymphocytic leukemia lymphocytes. Human lymphoblastic lines vary markedly in activity. In general, cell lines with B- and T-characteristics from patients with lymphoproliferative diseases had much lower activities than those of B-cell lines derived from normal subjects. The highest activity found was in a human myeloma line active in synthesis of an immunoglobulin light chain. The data indicate that gamma-glutamyl transpeptidase is a surface marker reflecting differentiation in normal and neoplastic cells.

Topics: Animals; B-Lymphocytes; Cell Differentiation; Cell Line; Cell Membrane; Concanavalin A; gamma-Glutamyltransferase; Guinea Pigs; Humans; Leukemia, Lymphoid; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Mice; Rats; Species Specificity; Spleen; T-Lymphocytes

The effects of phytohemagglutinin (PHA), concanavalin a (con A) and endotoxin on glucose oxidation by lymphocytes from normal individuals and patients with chronic lymphocytic leukemia (Cll) were demonstrated without using tissue culture techniques. PHA and CON A had no effect on glucose-6-14C oxidation, but stimulated glucose-1-14C oxidation by normal lmphocytes. This stimulation of glucose-1-14C oxidation occurred within 1 hour and could be prevented by colchicine. In contrast, endotoxin had no effect on either gucose-I-14C or glucose-6-14C oxidation by normal lymphocytes. Lymphocytes from five out of six Cll patients showed low basal rate of gucose-1-14C oxidation by the lymphocytes of these five patients.

Topics: 4-Chloromercuribenzenesulfonate; Aged; Colchicine; Concanavalin A; Drug Antagonism; Endotoxins; Female; Glucose; Humans; Lectins; Leukemia, Lymphoid; Lymphocytes; Magnesium; Middle Aged; Mitogens; Oxidation-Reduction; Stimulation, Chemical

The in vitro mitogen response of blood lymphoid cells from patients with chronic lymphocytic leukemia was observed to be low compared to lymphocytes from healthy subjects. However, highly responsive cells could be separated from the blood of such patients by a buoyant density centrifugation technique and such reactive cells were found to be enriched by treating the patients with chemotherapy or splenic irradiation.

Topics: Adult; Aged; Cell Separation; Chlorambucil; Concanavalin A; Cyclophosphamide; Female; Humans; Immunologic Techniques; In Vitro Techniques; Lectins; Leukemia, Lymphoid; Leukocyte Count; Lymphoma; Male; Middle Aged; Spleen; T-Lymphocytes

Lymphocytes from a patient with chronic lymphocytic leukaemia, in whom therapy was ineffective, were defined as thymus-derived (T) cells by membrane markers (sheep erythrocyte rosettes, complement rosettes, surface immunoglobulin). The lymphocytes responded weakly to two mitogens, phytohaemagglutinin and the calcium ionophore A23187, but not to concanavalin A. Cytogenetic studies of leukaemic cells from unstimulated and mitogen-stimulated cultures revealed an abnormal karyotype with 45 chromosomes and multiple rearrangements. The T cell variant of classical chronic lymphocytic leukaemia is relatively rare; additional reports are needed to determine if the clinical course is typically less benign than in the common B cell variety, or whether this patient simply represented a late, unresponsive phase of the disease.

Topics: Aged; Calcimycin; Cell Membrane; Chromosome Aberrations; Chromosome Disorders; Concanavalin A; Female; Genetic Variation; Humans; Karyotyping; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Mitogens; T-Lymphocytes

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

Topics: Aging; Animals; Cell Membrane; Concanavalin A; Histocompatibility Antigens; Lectins; Leukemia, Lymphoid; Lipopolysaccharides; Lymphocyte Activation; Mice; Mice, Inbred AKR; Mice, Inbred C3H; Species Specificity; T-Lymphocytes

The ultrastructure of the cell coat of normal and chronic lymphocytic leukaemic (CLL) blood lymphocytes has been compared by several cytochemical techniques. In CLL lymphocytes, there is no modification of Concanavalin A labelling, whereas a difference of stainability is observed with at least one of the three cationic dyes used: colloidal iron, colloidal thorium, or ruthenium red, signifying a modification of anionic reactive sites. This physico-chemical surface anomaly is discussed with regard to other biochemical and electrophoretic results given by other authors.

Topics: Cell Membrane; Colloids; Concanavalin A; Humans; Iron; Leukemia, Lymphoid; Lymphocytes; Ruthenium Red; Staining and Labeling; Thorium

Surface antigenic specificities of human thymus-derived (T) lymphocytes were studied by cytotoxicity tests using a heterologous rabbit anti-human thymus serum. This serum showed higher cytotoxic titres on thymocytes by comparison with peripheral lymphocytes. After proper absorption the antiserum was non-toxic for chronic lymphatic leukaemia cells, but lysed the majority of thymocytes. It also lysed some of peripheral lymphocytes, corresponding to those lymphocytes which bound sheep erythrocytes (E) but not erythrocyte-antibody-complement complexes (EAC). Pretreatment of lymphocytes with the absorbed antiserum and complement completely abrogated rosette formation with E but spared EAC-binding lymphocytes. It also eliminated their reactivity to phytohaemagglutinin and concanavalin A. These findings indicate that the absorbed serum causes selective lysis of T cells. The results obtained from quantitative absorption studies suggest that a certain loss of T-cell antigens is brought about during the differentiation of thymocytes into peripheral T cells.

Topics: Animals; Antilymphocyte Serum; Cell Membrane; Child; Complement System Proteins; Concanavalin A; Cytotoxicity Tests, Immunologic; Epitopes; Erythrocytes; Humans; Immune Adherence Reaction; In Vitro Techniques; Lectins; Leukemia, Lymphoid; Rabbits; Receptors, Antigen, B-Cell; T-Lymphocytes; Thymidine

The effect of Herpesvirus saimiri (HVS) infection on the response of owl monkey lymphocytes to general mitogens was examined during the development of neoplastic disease. The reactivity of the lymphocytes was then correlated with the clinical condition of the infected monkeys and the content of virus rescued from the peripheral blood lymphocytes (PBL). Eight monkeys developed lymphoma which, in six monkeys, was accompanied by lymphocytic leukemia. All animals that died of HVS-induced neoplasia consistently showed a lack of mitogenic response to phytohemagglutinin and concanavalin A. The response to pokeweed mitogen, while always reduced, was generally less markedly affected than the response to the other two mitogens. Lymphocytes from five of the leukemic animals demonstrated an elevated level of spontaneous DNA synthesis in culture late in the disease. This increased spontaneous DNA synthesis tended to correlate with the rescue of HVS from the PBL as demonstrated by the infective center assay. Although mitogenic hyporesponsiveness corresponded with HVS rescue from PBL in six of nine monkeys, the impairment of normal lymphocyte responsiveness sometimes preceded virus recovery.

Topics: Animals; Concanavalin A; DNA; Herpesviridae; Herpesviridae Infections; Herpesvirus 2, Saimiriine; Immunity, Cellular; Lectins; Leukemia, Lymphoid; Leukocyte Count; Lymphocytes; Lymphoma; Mitogens; Neoplasms, Experimental

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

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

Human peripheral blood lymphocytes (PBL) were evaluated by their responses to phytohemmagglutinin (PHA-P), concanavallin A (con-A), and pokeweed mitogen (PWM), both before and after treatment with an antiserum against human thymic lymphocyte antigens (HTLA) that had been made T-cell-specific by multiple absorptions with immunoglobulin EAC-positive lymphoblast cell lines (B cells). Cells treated with HTLA were examined for their ability to react in a mixed lymphocyte culture (MLC) and to form killer cells in a cell-mediated lymphocytotoxicity (CML) system. Sensitized cells were also examined for their ability to respond to purified protein derivative (PPD) by blastogenesis, migration inhibitory factor release (MIP), and lymphotoxin (LT) production, both before and after treatment with HTLA and complement. The HTLA was in itself highly stimulatory to PBL. However, with the addition of complement and subsequent cell destruction, a marked decrease in its stimulatory response was noted. PBL treated with HTLA and complement exhibited marked inhibition of responsiveness to con-A with little decrease in PHA-P -OR PWM stimulation except at very high concentration of HTLA. MLC reaction was inhibited only when responder cells were treated with HTLA + C'. Treatment of stimulator cells with HTLA + C' did not significantly alter the MLC response. The HTLA + C'-treated cells failed to form killer cells in the CML reaction and inhibited PPD-induced blasto-genesis from PPD-sensitized individuals; however, treatment of sensitized cells with HTLA + C' had little effects on the release of MIF and LT. It is suggested that subpopulations of T-cells carry surface antigens that bind with this specific antisera, and that the con-A-responsive cells, the responder cells in the MLC, and killer T-cells comprise a separate subset from cells responding to PHA-P or PWM, OR THE MIF-and LT-producing cells.

Topics: Animals; Antilymphocyte Serum; Cell Line; Complement System Proteins; Concanavalin A; Cytotoxicity Tests, Immunologic; Histocompatibility Testing; Humans; Immune Adherence Reaction; Immunoglobulins; In Vitro Techniques; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphotoxin-alpha; Macrophage Migration-Inhibitory Factors; Mitogens; Plant Extracts; Rabbits; T-Lymphocytes; Tuberculin

Rosette formation between human thymus-derived (T) lymphocytes and sheep red blood cells (SRBC) was inhibited by T cell specific antisera but not by sera directed against other lymphocyte antigens. We conclude that the SRBC receptor is closely linked to T specific antigens.

Topics: Adsorption; Animals; Antibody Specificity; Antilymphocyte Serum; B-Lymphocytes; Binding Sites, Antibody; Cell Membrane; Concanavalin A; Erythrocytes; Fluorescent Antibody Technique; Glycoproteins; Horses; Humans; Immune Adherence Reaction; Immunosuppression Therapy; Isoantigens; Leukemia, Lymphoid; Molecular Weight; Neuraminidase; Sheep; T-Lymphocytes; Thymus Gland

Cell-line lymphocytes contained considerably more nonhistone chromatin protein than normal lymphocytes. The overall electrophoretic pattern of the nonhistone chromatin proteins from nonreplicating cell types with predominantly condensed chromatin (normal lymphocytes and chronic lymphocytic leukemia lymphocytes) was similar, as it was between replicating cell types with predominantly decondensed chromatin (mitogen-stimulated normal lymphocytes and cell-line lymphocytes), although all four cell types differed qualitatively. A "heavy" chromatin fraction from cell-line lymphocytes could be distinguished from a "light" fraction and had a pattern similar to that from cells of the condensed chromatin type. Normal lymphocytes contained two acid-soluble chromatin polypeptieds not found in cell-line lymphocytes.

Topics: Cell Fractionation; Cell Line; Chromatin; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Humans; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Neoplasm Proteins; Peptides; Proteins; Sodium Dodecyl Sulfate

Topics: Aged; Agglutination Tests; Cell Membrane; Concanavalin A; Female; Hodgkin Disease; Humans; Immune Adherence Reaction; Immunity, Cellular; Leukemia, Lymphoid; Lymphocytes; Male; Middle Aged; Remission, Spontaneous; Stimulation, Chemical; Vinblastine

Topics: Animals; Antibodies, Viral; Concanavalin A; Disease Models, Animal; Fluorescent Antibody Technique; Haplorhini; Herpesviridae; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Lymphoma; Mitogens; Monkey Diseases; Oncogenic Viruses

Topics: Agglutination; Agglutination Tests; Binding Sites, Antibody; Bone Marrow; Bone Marrow Cells; Cell Separation; Cells, Cultured; Concanavalin A; Iodine Radioisotopes; Leukemia, Lymphoid; Lymphocytes; Sodium

Topics: Adult; Aged; Binding Sites, Antibody; Cell Membrane; Concanavalin A; Dose-Response Relationship, Drug; Female; Hemagglutination Tests; Humans; Leukemia, Lymphoid; Lymphocytes; Male; Methods; Middle Aged; Stimulation, Chemical; Tritium

Topics: Blood Sedimentation; Carbon Radioisotopes; Cell Nucleus; Cell Survival; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Drug; Erythrocytes; Glutathione; Humans; Lectins; Leukemia, Lymphoid; Lymphocytes; Mutagens; Plant Extracts; RNA; Stimulation, Chemical; Thymidine; Tritium; Uridine

Topics: ABO Blood-Group System; Acetamides; Animals; beta-Aminoethyl Isothiourea; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Separation; Complement System Proteins; Concanavalin A; Cysteine; Cytotoxicity Tests, Immunologic; Dithiothreitol; Dose-Response Relationship, Drug; Erythrocytes; Glutathione; Hemagglutination Tests; Hemoglobinuria, Paroxysmal; Humans; Hydrogen-Ion Concentration; Hypersplenism; Immune Adherence Reaction; Isoantibodies; Leukemia, Lymphoid; Lupus Erythematosus, Systemic; Lymphocytes; Mercaptoethanol; Microscopy, Electron, Scanning; Sheep; Sulfhydryl Compounds

The blood lymphocytes from a case of prolymphocytic leukaemia were subjected to a battery of different tests in order to establish as certainly as possible their T or B cell type of origin. The results of the tests for surface markers indicated the T-cell origin of the leukaemic cells in this patient, and this provided a good opportunity to determine the participation of T cells in the various tests proposed for measuring human lymphocyte function.

Topics: Aminosalicylic Acids; Animals; Antibodies; Autoradiography; Complement System Proteins; Concanavalin A; Electrophoresis; Erythrocytes; Female; Fluorescent Antibody Technique; Humans; Immune Adherence Reaction; Immune Sera; Immunoglobulins; Immunologic Techniques; Lectins; Leukemia, Lymphoid; Lymphocytes; Microscopy, Electron; Middle Aged; Mitogens; Sheep; T-Lymphocytes; Thymidine; Tritium

Topics: Animals; Antilymphocyte Serum; Azathioprine; B-Lymphocytes; Bacterial Infections; Concanavalin A; Dermatitis, Atopic; Graft Rejection; Hodgkin Disease; Humans; Immunosuppressive Agents; Kidney Transplantation; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Prednisone; Rabbits; Renal Dialysis; Sarcoidosis; T-Lymphocytes; Transplantation, Homologous

Topics: Cell Line; Cell Nucleus; Cell Survival; Cell Wall; Cells, Cultured; Concanavalin A; Cytotoxicity Tests, Immunologic; Humans; Lectins; Leukemia, Lymphoid; Liver; Lymphocytes; Mitogens; Plant Extracts

Topics: Cell Transformation, Neoplastic; Concanavalin A; Hodgkin Disease; Humans; Immune Adherence Reaction; Immunity, Cellular; Lectins; Leukemia, Lymphoid; Lymphoma, Non-Hodgkin; Mitogens; T-Lymphocytes

Topics: B-Lymphocytes; Cell Nucleus; Cells, Cultured; Concanavalin A; Cytoplasm; DNA; Humans; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Microscopy, Electron; Mitochondria; T-Lymphocytes; Thymidine; Tritium

Topics: Autoradiography; Blood; Chronic Disease; Concanavalin A; Culture Media; Female; Fluorescent Antibody Technique; Humans; Immunoglobulin A; Inclusion Bodies; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Microscopy, Electron; Middle Aged; Stimulation, Chemical; Thymidine; Tritium

Topics: Animals; Cells, Cultured; Complement System Proteins; Concanavalin A; DNA; Fluorescent Antibody Technique; Hodgkin Disease; Humans; Immunoglobulins; Kidney Diseases; Lectins; Leukemia, Lymphoid; Rabbits; T-Lymphocytes; Tritium

Topics: Acid Phosphatase; Cell Nucleus; Concanavalin A; Cytoplasm; Histocytochemistry; Humans; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Lysosomes; Microscopy, Electron

Topics: Acid Phosphatase; Cells, Cultured; Concanavalin A; Humans; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Lysosomes; Microscopy, Electron; Mitogens; Thymidine; Tritium

Topics: Agammaglobulinemia; Cell Fractionation; Cells, Cultured; Centrifugation, Density Gradient; Concanavalin A; Humans; Lectins; Leukemia, Lymphoid; Lymphocytes; Mitogens; Mitosis; Serum Albumin, Bovine; Thymidine; Tritium

Topics: Concanavalin A; Culture Techniques; Humans; Lectins; Leukemia, Lymphoid; Lymphocytes; Lysosomes; Mitogens

Topics: Cells, Cultured; Chronic Disease; Concanavalin A; Humans; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Thymidine; Tritium

Topics: Agglutination; Animals; Avian Sarcoma Viruses; Cell Line; Cell Transformation, Neoplastic; Chick Embryo; Concanavalin A; Erythrocytes; Humans; Lectins; Leukemia, Lymphoid; Liver; Lymphocytes; Mice; Neoplasms, Experimental; Neuraminidase; Plant Lectins; Rats; Spleen; Triticum; Trypsin

Essentially no specific binding sites for insulin are detected in small lymphocytes freshly isolated from human blood. Insulin-binding sites appear on the lymphocyte surface during transformation in vitro with concanavalin A, and the number of these receptors increases sharply to reach a maximum between 24 and 46 hr after exposure to the mitogen. The appearance of de novo binding sites for insulin coincides with the increase in [(3)H]thymidine uptake into nuclear DNA and clearly precedes the appearance of enlarged, morphologically transformed cells. No changes in insulin-binding are detected in unstimulated control cultures. A maximum of about 350 molecules of insulin can bind per transformed lymphocyte, while less than six insulin molecules bind to an untransformed cell. Circulating human leukemic lymphoblasts bind about as much insulin as the lymphocytes transformed in vitro. Giant, polynucleated, transformed lymphocytes cultured in the presence of cytochalasin B bind about 10 times more insulin than transformed lymphocytes, which is in harmony with a 10-fold increase in cell-surface area in these cells. Specific binding of insulin is a saturable process in transformed lymphocytes but not in the untransformed cells. In transformed cells, [(125)I]-insulin is displaced by as little as 2 ng/ml of native insulin, while in untransformed cells no significant displacement is observed with native insulin. Digestion of transformed cells with phospholipase C (EC 3.1.4.3.) enhances the specific binding of [(125)I]insulin 3-fold, but no effect occurs with untransformed cells. These observations indicate a possible functional role of insulin and of adenylate cyclase in cell growth and division.

Topics: Binding Sites; Cell Differentiation; Cell Division; Cells, Cultured; Clostridium perfringens; Concanavalin A; DNA; Humans; Insulin; Iodine Isotopes; Kinetics; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Mycotoxins; Phospholipases; Protein Binding; Receptors, Drug; Thymidine; Tritium

Topics: Animals; Antibody Formation; Binding Sites; Bone Marrow; Bone Marrow Cells; Carbon Isotopes; Cells, Cultured; Complement System Proteins; Concanavalin A; Fluorescent Antibody Technique; Guinea Pigs; Histocompatibility; Immune Adherence Reaction; Immune Sera; Immunoelectrophoresis; Immunoglobulins; Lectins; Leukemia, Experimental; Leukemia, Lymphoid; Lymph Nodes; Lymphocytes; Lysine; Thymidine; Thymus Gland; Tritium

Topics: Adult; Aged; Animals; Antilymphocyte Serum; B-Lymphocytes; Binding Sites; Cell Membrane; Concanavalin A; Erythrocytes; Female; Fluorescent Antibody Technique; Humans; Immune Adherence Reaction; Immunoglobulin G; Immunoglobulin M; Isoantigens; Leukemia, Lymphoid; Leukocyte Count; Lymphocytes; Lymphoma, Non-Hodgkin; Male; Middle Aged; Sheep; T-Lymphocytes

We have identified two populations of human lymphocytes differing in responsiveness to the plant mitogen concanavalin A (Con-A). When peripheral blood lymphocytes are passed through a nylon column a population of lymphocytes highly responsive to Con-A adheres to the fibers while a second population of cells relatively unresponsive to Con-A emerges from the column. The untreated peripheral blood lymphocytes are termed "unfiltered" cells while the lymphocytes which pass through the column are termed "filtered" cells. Under standard assay conditions the Con-A-stimulated DNA synthesis is 6.5-fold greater, and the percentage blast formation is four-to fivefold greater in the unfiltered than in the filtered population. Mixing unfiltered with filtered cells fails to induce responsiveness in the latter indicating that a "helper" cell is not involved. The failure of filtered cells to respond to Con-A is specific for that mitogen since both populations respond nearly equally to erythroagglutinating phytohemagglutinin (E-PHA) and the poke weed mitogen (PWM). Binding studies with Con-A-(131)I demonstrate that the unfiltered population possesses approximately three times as many Con-A receptor sites per cell as the filtered cells, although both cell populations bind the mitogen with the same affinity (apparent association constant [K] of 1.67 x 10(6)m(-1)). The relationship between Con-A binding and lymphocyte activation was determined by measuring the effect on DNA synthesis of incubating the two lymphocyte populations with increasing amounts of Con-A. The concentration of Con-A required for half-maximal stimulation of DNA synthesis was 5-14 times greater for the filtered cells. However in the presence of very high Con-A concentrations the filtered cells achieved a maximal rate of DNA synthesis approaching that of the unfiltered population. These data implicate the decreased number of Con-A receptor sites on the filtered cells in their failure to respond to low concentrations of Con-A. A crucial event in the activation of lymphocytes by plant mitogens may be the binding of a critical number of the mitogen molecules to the cell surface.

Topics: Binding Sites, Antibody; Cell Fractionation; Cell Membrane; Concanavalin A; DNA; Humans; Iodine Isotopes; Lectins; Leukemia, Lymphoid; Lymphocyte Activation; Lymphocytes; Thymidine; Tritium

Topics: Chromatography; Concanavalin A; Culture Techniques; DNA; Humans; Iodine Isotopes; Lectins; Leukemia, Lymphoid; Lymphocytes; Micropore Filters; Mitogens; Mitosis; Stimulation, Chemical; Thymidine; Tritium

Topics: Animals; Binding Sites; Cells, Cultured; Concanavalin A; DNA; Humans; Lectins; Leukemia, Lymphoid; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Male; Niobium; Rats; RNA; Thymidine; Tritium; Uridine