concanavalin-a and Lymphoma

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: Animals; Antigens, Surface; Biophysical Phenomena; Biophysics; Cell Membrane; Concanavalin A; Epitopes; Fibroblasts; H-2 Antigens; Humans; Lymphocytes; Lymphoma; Mice; Mice, Inbred A; Receptors, Concanavalin A; Rotation

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

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

Stress alters the neuroendocrine system, immunity, and cancer. Although the classic stress hormones are glucocorticoids and catecholamines, thyroid hormones have also been related to stress. We recently reported that chronic restraint stress impairs T-cell mediated immunity and enhances tumor growth in mice.. To study the participation of these hormones on the stress-induced alterations of the immune function and lymphoma growth, mice were subjected to acute or chronic stress, with or without thyroxin supplementation. Hormone levels, immune status, and cancer progression were evaluated.. Differential endocrine alterations were observed in response to acute and chronic stress. Although corticosterone and noradrenaline levels were increased by acute stress, they were restored after prolonged exposure to the stressor. Instead, thyroid hormone levels were only reduced in chronically stressed animals in comparison with control subjects. Correlating, chronic but not acute stress impaired T-cell reactivity. Thyroxin replacement treatment of chronic restraint stress-exposed mice, which restored the euthyroid status, reversed the observed reduction of T-cell lymphoproliferative responses. Moreover, therapeutic thyroid replacement also reversed the alterations of lymphoma growth induced by chronic stress in syngeneic mice bearing tumors as well as Interleukin-2 production and specific cytotoxic response against tumor cells. Finally, we found that the isoforms theta and alpha of the protein kinase C are involved in these events.. These results show for the first time that thyroid hormones are important neuroendocrine regulators of tumor evolution, most probably acting through the modulation of T-cell mediated immunity affected by chronic stress.

Topics: Animals; Cell Proliferation; Concanavalin A; Corticosterone; Disease Models, Animal; Disease Progression; Female; Flow Cytometry; Lymphoma; Mice; Mice, Inbred BALB C; Mitogens; Norepinephrine; Protein Kinase C; Restraint, Physical; Stress, Psychological; T-Lymphocytes; Thymidine; Thyroid Hormones; Thyroxine; Tritium

It has been reported that concentrations of neopterin in the urine are changed according to the host immunological conditions. In the present study, we measured urinary concentration of neopterin in patients with malignant hematological disorders and investigated the relationship between urinary neopterin levels and laboratory indices for cellular immunity. Urine neopterin levels were correlated with serum sIL-2R levels in the patients with malignant lymphoma, and inversely correlated with lymphocyte reactivity with ConA in the patients with acute myelocytic leukemia. However, no significant correlation was observed between urine neopterin levels and lymphocyte reactivity with phytohemagglutinin and pokeweed mitogen, CD4/8 ratio, CD56+ 16+ subset or serum IFN-gamma levels. In the patients with malignant lymphoma, parallel changes in serum sIL-2R and urine neopterin were observed. The presented results suggest that urine neopterin levels are related to the activation of T cells in malignant lymphoma.

Topics: Adult; Aged; Concanavalin A; Female; Humans; Leukemia, Myeloid, Acute; Lymphoma; Male; Middle Aged; Neopterin; Receptors, Interleukin-2

The effect of thyroid hormone on immune function is unclear. The influence of L-triiodothyronine on expression of the interleukin-2 receptor alpha chain by peripheral blood mononuclear cells from healthy volunteers and YT cells (an interleukin-2 independent natural killer-like cell line) was examined. Concanavalin A stimulation significantly (p<0.05 and p<0.01) increased soluble interleukin-2 receptor alpha chain production when mononuclear cells were cultured with triiodothyronine (1-100 nmol/l) for 3 days. The stimulatory effect of triiodothyronine on interleukin-2 receptor alpha chain expression was greater in the presence of concanavalin A (5 microg/ml) plus interleukin-2 (1 U/ml) than in the presence of concanavalin A alone. Triiodothyronine also significantly (p<0.01) increased interleukin-2 receptor alpha chain expression when YT cells were cultured for 2 days with interleukin-2 (1 U/ml), but did not influence receptor expression when YT cells were cultured with forskolin or 12-O-tetradecanoyl phorbol 13-acetate, potent activators of signal transduction. In conclusion, triiodothyronine may have an immunomodulatory effect by enhancing expression of the interleukin-2 receptor alpha chain on peripheral blood mononuclear cells in the presence of interleukin-2.

Topics: Cells, Cultured; Colforsin; Concanavalin A; Gene Expression; Humans; Killer Cells, Natural; Leukocytes, Mononuclear; Lymphoma; Receptors, Interleukin-2; Solubility; Tetradecanoylphorbol Acetate; Triiodothyronine; Tumor Cells, Cultured

Mice lacking beta2-microglobulin (beta2m- mice) express greatly reduced levels of MHC class I molecules, and cells from beta2m- mice are therefore highly sensitive to NK cells. However, NK cells from beta2m- mice fail to kill beta2m- normal cells, showing that they are self tolerant. In a first attempt to understand better the basis of this tolerance, we have analyzed more extensively the target cell specificity of beta2m- NK cells. In a comparison between several MHC class I-deficient and positive target cell pairs for sensitivity to beta2m- NK cells, we made the following observations: First, beta2m- NK cells displayed a close to normal ability to kill a panel of MHC class I-deficient tumor cells, despite their nonresponsiveness to beta2m- concanavalin A (Con A)-activated T cell blasts. Secondly, beta2m- NK cells were highly sensitive to MHC class I-mediated inhibition, in fact more so than beta2m+ NK cells. Thirdly beta2m- NK cells were not only tolerant to beta2m- Con A blasts but also to Con A blasts from H-2Kb-/Db- double deficient mice in vitro. We conclude that NK cell tolerance against MHC class I-deficient targets is restricted to nontransformed cells and independent of target cell expression of MHC class I free heavy chains. The enhanced ability of beta2m- NK cells to distinguish between MHC class I-negative and -positive target cells may be explained by increased expression of Ly49 receptors, as described previously. However, the mechanisms for enhanced inhibition by MHC class I molecules appear to be unrelated to self tolerance in beta2m- mice, which may instead operate through mechanisms involving triggering pathways.

Topics: Animals; Antigen Presentation; beta 2-Microglobulin; Concanavalin A; Crosses, Genetic; Cytotoxicity, Immunologic; Genes, MHC Class I; H-2 Antigens; Histocompatibility Antigen H-2D; Interferon-gamma; Killer Cells, Natural; Lung Neoplasms; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred BALB C; Mice, Knockout; Neoplasm Transplantation; Recombinant Proteins; Self Tolerance; T-Lymphocytes; Tumor Cells, Cultured

Overexpression of the bcl-2 oncogene in the lymphoid compartment of transgenic mice prolongs the lifespan of lymphocytes and leads to a low incidence of lymphomas at later age. Transgenic mice carrying a mutated T-cell receptor lacking the variable domain (deltaV-TCRbeta) suffer from lymphocyte depletion and are highly predisposed to lymphoma development. We intercrossed Bcl-2-Ig and deltaV-TCRbeta transgenic mice to assess whether Bcl-2 could synergize with deltaV-TCRbeta in tumorigenesis as reported previously for other oncogenes. Surprisingly, bitransgenic deltaV-TCRbeta; bcl-2-Ig mice showed a reduction in the incidence of lymphomas. Analyses of prelymphomatous mice showed that Bcl-2 restored some of the phenotypic aberrations caused by the deltaV-TCRbeta transgene in the lymphoid compartment. The inhibitory activity of Bcl-2 on deltaVTCRbeta-induced lymphomagenesis was not observed when both transgenes were crossed into the RAG-1-/- background suggesting an important role for more mature lymphocytes in this phenomenon. These results show that, depending on the specific conditions, overexpression of Bcl-2 can both promote as well as impair lymphoma development.

Topics: Animals; Antigens, CD; B-Lymphocytes; Cell Survival; Concanavalin A; Crosses, Genetic; Female; Interleukin-7; Lipopolysaccharides; Lymphoma; Macrophage Colony-Stimulating Factor; Male; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Mice, Transgenic; Phenotype; Proto-Oncogene Proteins c-bcl-2; Receptors, Antigen, T-Cell, alpha-beta; Sex Factors; Spleen; Survival Rate; T-Lymphocytes; Time Factors

We have evaluated the production of PRL by human peripheral mononuclear cells (PBMNC) from normal subjects and patients with systemic lupus erythematosus (SLE). Conditioned medium prepared from basal and Con-A-stimulated PBMNC was assessed for the presence of PRL-like by its ability to stimulate growth of PRL-responsive Nb2 rat lymphoma cells. In the presence or absence of Con-A, SLE PBMNC secrete significantly higher (P < 0.001) amounts of bioactive PRL-like species than normal cells. Growth of Nb2 cells by conditioned medium was inhibited with specific antiserum to human PRL. Western blotting using a polyclonal antibody to human PRL revealed a single 60-kDa PRL-like species in both normal and SLE PBMNC extracts, the immunoreactivity of which was preferentially found in SLE subjects. With the use of reverse transcription-PCR an expected 633-bp band was observed, and its similarity to pituitary PRL was further confirmed by Southern blot analysis with human PRL complementary DNA as a probe. We conclude that a high molecular mass PRL-like species is synthesized and secreted by PBMNC, and patients with SLE have an increased secretion of lymphocyte-derived PRL-like material.

Topics: Adolescent; Adult; Animals; Biological Assay; Blotting, Southern; Blotting, Western; Cells, Cultured; Concanavalin A; Culture Media, Conditioned; Female; Gene Expression; Humans; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Lymphoma; Molecular Weight; Polymerase Chain Reaction; Prolactin; Rats; RNA, Messenger

Thymocyte apoptosis was assessed by counting apoptotic bodies with flow cytometry (FCM) and measuring DNA fragmentation with fluorescence spectrophotometry (FSP). J-shaped dose-response curves were obtained after both whole-body irradiation (WBI) of mice and in vitro irradiation of EL4 cells with doses ranging from 0.025 to 4 Gy X-rays. There was a significant reduction of apoptosis rate to below control level with doses within 0.2 Gy, and a dose-dependent increase in apoptosis with doses above 0.5 Gy. When thymocytes were cultured 24 h after WBI with 75 mGy X-rays in complete RPMI 1640 medium, a reduction in apoptosis was observed in the course of incubation for 72 h, and the presence of Con A in the medium accentuated this reduction in a dose- and time-dependent manner. The implications of these observations and the possible molecular mechanisms for future studies are proposed.

Topics: Animals; Apoptosis; Cell Cycle; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Radiation; Lymphoma; Male; Mice; Mice, Inbred Strains; Thymus Gland; Tumor Cells, Cultured; Whole-Body Irradiation; X-Rays

Several studies have demonstrated that dietary nucleotides play a role in maintaining T-cell dependent immunity. In this work, we investigated the effects of nucleotide supplementation of a nucleotide-free diet (NFD) on some immunity parameters in BALB/c mice. Twenty day old mice were maintained on diets for 30 days prior to use in experiments. The addition of nucleotide mixtures to NFD resulted in an increase in the response of hemolytic IgG-forming cells induced by previous immunization with sheep erythrocytes. When NFD was supplemented with single nucleotides, AMP, GMP, or UMP increased the IgG response, whereas CMP and IMP were without effect. GMP was the only nucleotide that increased the hemolytic IgM-forming cell response. Neither the contact hypersensitivity response to dinitrofluorobenzene nor the time of death after transplantation of a syngenic lymphoma was modified by nucleotide addition to NFD. The in vitro proliferative response of splenocytes to LPS was not affected by nucleotide supplementation of NFD, but the ConA-driven proliferative response was increased in mice fed NFD supplemented with nucleotide mixtures or with UMP. These data show that dietary mononucleotides stimulated at least some T-cell dependent immunity mechanisms. Moreover, these stimulatory effects may be obtained by supplementing a nucleotide-free diet with a mixture in which mononucleotides are at the same levels as in murine breast milk.

Topics: Animals; B-Lymphocytes; Cells, Cultured; Concanavalin A; Dermatitis, Contact; Dinitrofluorobenzene; Erythrocytes; Female; Food, Fortified; Immunoglobulin G; Lipopolysaccharides; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred BALB C; Nucleotides; Sheep; Transplantation, Isogeneic

The prolactin (PRL)-like activity released into synthetic culture medium from concanavalin-A (Con-A)-stimulated mouse splenocytes was bioassayed using 3H thymidine incorporation into Nb2 lymphoma cells. At low cell density (1.0 x 10(6) cells/ml medium) Con-A-stimulated splenocytes from Balb/c mice released more PRL-like activity than did splenocytes from normal C3H/HeJ mice. This difference was maintained with animals of either sex, and for animals of different ages (1, 3 and 5 months). The strain difference was observed both when splenocytes were co-cultured with Nb2 cells, and when postcultured medium from splenocytes was tested. Con-A-stimulated splenocytes also released a substance into the medium which was inhibitory to PRL-induced Nb2 cell proliferation. The concentration of the inhibitory substance in the medium was related to the number of splenocytes in culture and was similar in splenocyte media preparations from both strains of mice. Polyclonal antisera to bovine PRL, rat PRL, mouse placental lactogen II, and mouse PRL did not neutralize the effect of Con-A-stimulated splenocytes to induce Nb2 cell proliferation. Bioassay of the partially purified PRL-like activity found in postcultured medium indicated that the dose-dependent induction of Nb2 cell proliferation was parallel with that of purified mPRL. Using two mouse PRL antisera (Sinha and Talamantes #118) in Western blot analyses, no antibody binding was observed to the partially-purified splenocyte PRL-like material. The data from the antisera neutralization experiments and the SDS/PAGE Western blot analyses clearly indicated that the PRL-like activity produced by Con-A-stimulated splenocytes has little homology with pituitary PRL. Further studies are required to establish the precise molecular identity of the PRL-like activity from Con-A-stimulated splenocytes.

Topics: Animals; Blotting, Western; Cell Division; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Drug; Female; Immune Sera; Lymphoma; Male; Mice; Mice, Inbred BALB C; Prolactin; Spleen; Thymidine

The production of a prolactin (PRL)-like substance by mitogen-stimulated immunocompetent cells has been reported previously for a number of species. The Snell dwarf mouse has a deficiency in thyrotropin (TSH), growth hormone (GH) and prolactin (PRL) as a result of a defect in the pituitary Pit-1 promoter. Since the gene for PRL is present in the dwarf mouse pituitary but not activated it was of interest to determine whether a similar deficiency existed for splenocytes from the dwarf animal. Irradiated splenocytes from dwarfs and normal littermates were cocultured in synthetic AIM-V medium with Nb2 cells and stimulated with concanavalvin A (Con-A). The 3H thymidine incorporation into Nb2 cells in cocultures was quantitated by the addition of mouse PRL to Nb2 cells alone. Splenocytes from dwarf mice produced significantly less PRL-like activity (p < 0.02) than did splenocytes from normal animals. The administration of thyroxine (T4) to dwarf mice increased body weight (BW) gain and the number of splenocytes/g BW. The administration of recombinant bovine GH but not recombinant bPRL further increased body weight gain over T4 alone but neither pituitary hormone had any additional effect on the number of splenocytes/g BW over that noted for T4 alone. Prolactin and GH alone had no effect on splenocyte numbers/g BW. The decreased production of PRL-like activity in the dwarf mouse was not altered by either GH or PRL injection. The injection of T4 alone and in combination with pituitary hormones increased the production of PRL-like activity by dwarf splenocytes to values similar to that observed for normal animals.

Topics: Animals; Body Weight; Cells, Cultured; Concanavalin A; Drug Interactions; Female; Growth Hormone; Lymphoma; Male; Mice; Mice, Inbred C3H; Prolactin; Spleen; Stimulation, Chemical; Thyroxine; Tumor Cells, Cultured

Class I molecules of the MHC affect target cell sensitivity to NK cell repertoire. In the mouse, absence of MHC class I molecules on target cells is associated with an increased susceptibility to NK cell-mediated lysis, while syngeneic class I molecules confer protection. In contrast to the protective role of syngeneic class I molecules, less is known about the interaction between murine NK cells and allogenic class I MHC molecules. In theory, such could either be triggering, inert, or inhibitory. To directly address the role of allogeneic class I in interaction with NK cells of the mouse, a panel of polyclonal allogeneic murine NK cells were exposed to H-2b class I positive or negative target cells, and susceptibility to lysis was assessed. For all effectors studied, regardless of H-2 haplotype or genetic background, a preferential killing of class I-deficient targets was observed. This pattern was observed in vitro with tumor as well as lymphoblast targets, and in vivo in rapid elimination studies of radiolabeled tumor cells. The results demonstrate that protection from murine NK cell-mediated lysis can be conferred by the expression of allogeneic class I molecules. No evidence for a triggering effect caused by the expression of allogeneic class I molecules was observed. The data are discussed in relation to current models on NK cell/MHC class I interactions, alloreactivity mediated by NK cells, and the role of NK cells in allogeneic graft rejection responses.

Topics: Animals; Cell Death; Concanavalin A; Cytotoxicity, Immunologic; H-2 Antigens; Haplotypes; Histocompatibility Antigens Class I; Immunotherapy, Adoptive; Killer Cells, Natural; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred Strains; T-Lymphocytes; Tumor Cells, Cultured

Rat Nb 2 lymphoma cells have been widely used to bioassay human growth hormone and many species of prolactin. Because their morphologic characterization suggests a T-cell lineage, Nb 2 cells were examined for their response to the T-cell mitogens concanavalin A, pokeweed mitogen, and phytohemagglutinin P. As expected, a dose-response to rat prolactin was observed; however, attempts to induce proliferation using the conventional T-cell mitogens failed at concentrations normally stimulatory for rat primary lymphocytes. Moreover, when Nb 2 cells were simultaneously incubated with lectin plus a suboptimal concentration of prolactin, a dose-dependent suppression of the stimulatory effects of prolactin was observed with phytohemagglutinin P and pokeweed mitogen, although not with concanavalin A. Culture medium of prolactin-stimulated Nb 2 cells also contained a factor which inhibited normal rat lymphocyte activation by concanavalin A. The factor did not block induction of the IL-2 receptor and proliferation of IL-2-dependent CTLL-2 cells could be restored by exogenous IL-2. Because Nb 2 cells evolved from a lactogen-dependent lymph node tumor, these results may have implications for further understanding the role of pituitary hormones, particularly prolactin, in the immune response to hormone-dependent tumor progression.

Topics: Animals; Cell Division; Concanavalin A; Humans; Interleukin-2; Lymphocyte Activation; Lymphoma; Mitogens; Prolactin; Rats; Spleen; Tumor Cells, Cultured

We have assessed several age-sensitive indicators of immune status in young (i.e., 6 to 11-month-old) mice of a genetically heterogeneous population to see if these varied in parallel and to determine if one or more of the status indices predicted life span or cancer incidence. We report that the number of memory (i.e., CD44hi) T cells within the CD8 subset is correlated with number of memory cells in the CD4 population, and inversely correlated with the number of naive (i.e., CD45RBhi) CD4 cells at both 6 and 11 months of age, suggesting that the conversion of naive to memory cells may occur at similar rates in both T cell subsets. Mice that ranked high in the proportion of memory T cells (within the CD4 and CD8 pools) at 6 months of age tended to retain their ranking at 11 months, suggesting that the pace or extent of memory cell formation may be a consistent trait that distinguishes mice at least within a genetically heterogeneous population. Mice that at 6 months of age exhibited high levels of CD4 or CD8 memory T cells, low levels of naive CD4 cells, or low levels of T cells able to proliferate in response to Con A and IL-2 were found to be significantly more likely than their littermates to die within the first 18 months of life. Cases of follicular cell lymphoma, lymphocytic and lymphoblastic lymphoma, and hepatic hemangiosarcoma were seen within the group of mice dying at early ages.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Aging; Animals; Carrier Proteins; Cause of Death; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Concanavalin A; Female; Hyaluronan Receptors; Immunologic Deficiency Syndromes; Immunologic Memory; Interleukin-2; Longevity; Lymphocyte Activation; Lymphoma; Male; Mice; Mice, Inbred AKR; Mice, Inbred C57BL; Mice, Inbred DBA; Mice, Inbred Strains; Phenotype; Receptors, Cell Surface; Receptors, Lymphocyte Homing; T-Lymphocyte Subsets

The influence of co-cultures of irradiated mouse thymocytes, splenocytes, and mesenteric lymph node cells on the proliferation of Nb2 cells in synthetic medium were studied. Irradiated thymocytes with or without the addition of ovine prolactin (oPRL) decreased the incorporation of 3H-thymidine into Nb2 cells. Irradiated splenocytes and lymph node cells when co-cultured with Nb2 cells and stimulated with concanavalin A (Con-A) induced Nb2 cell proliferation. Lipopolysaccharide stimulation of irradiated splenocytes and lymph node cells, however, did not alter Nb2 cell proliferation. Isolated thioglycolate-induced peritoneal macrophages stimulated Nb2 cells to incorporate 3H-thymidine while IFN-gamma-stimulated macrophages decreased PRL-induced Nb2 cell proliferation when isolated 48 but not 24 hr before co-culture. The addition of a mouse PRL antiserum to macrophage/Nb2 cell co-cultures did not alter the proliferative activity induced by macrophages. The preliminary data presented indicate that Con-A-stimulated, irradiated splenocytes and lymph node cells, and isolated peritoneal macrophages secrete a PRL-like activity. The PRL-like activity of macrophages, however, does not appear to be of mouse origin, and it is suggested that macrophages, which have surface PRL receptors, sequestered PRL from the fetal calf serum in the medium used to isolate them and release this PRL when co-cultured.

Topics: Animals; Cell Division; Cell Line; Concanavalin A; Cytokines; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukins; Lipopolysaccharides; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Lymphoma; Macrophages, Peritoneal; Mice; Mice, Inbred BALB C; Prolactin; Rats; Recombinant Proteins; Sheep; Spleen; T-Lymphocytes; Thymidine; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Natural polyamines (putrescine, spermidine, and spermine) are ubiquitous cellular cations that play an important role in cell proliferation and differentiation. Ornithine decarboxylase is the first and a rate-limiting enzyme in the biosynthesis of polyamines. Polyamine depletion using DL-alpha-difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase, has been shown to suppress cell growth in a variety of settings, including those of tumor and lymphocyte proliferation. The objective of the present investigation was to examine the inhibitory effects of DFMO on a variety of murine in vitro immune responses, including lymphocyte proliferation in response to T-cell mitogen (concanavalin A), B-cell mitogen (lipopolysaccharide), and alloantigen as well as cytotoxicity. DFMO-mediated inhibition of cell proliferation in these cases correlated with depletion of intracellular polyamines. The inhibitory effects of DFMO were reversed by polyamine repletion with putrescine. Putrescine also reversed the growth-inhibitory effects of DFMO on 4 tumor cell lines that we tested: 28-13-3S, YAC-1, P-815, and K562. However, putrescine homologues exhibited a differential effect in preventing DFMO-mediated inhibition of cell growth in normal lymphocytes and cancer cell lines. Only putrescine homologues containing a shorter methylene chain were effective in preventing the growth-inhibitory action of DFMO on normal immune response. In contrast, only the longer chain homologue 1,5-diaminopentane overcame the effect of DFMO on tumor cell growth. These findings suggest that supplementation with selected polyamine homologues may sustain normal immune response in DFMO-treated individuals while effectively suppressing malignant cell growth. The potential clinical relevance of these observations is discussed.

Topics: Animals; Cell Division; Cell Line; Concanavalin A; Cytotoxicity, Immunologic; Eflornithine; Isomerism; Lymphocyte Activation; Lymphoma; Male; Mast-Cell Sarcoma; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Polyamines; Putrescine; Spermidine; Spermine

Binding of the lectin Con A to its ligand on the cell surface of normal circulating lymphocytes induces capping in 28-32% of these cells. This Con A cap formation is markedly decreased in malignant cells from the human hematopoietic system. Among others, the human lymphoma Daudi cell line exhibit a cap formation with Con A in only 5-10% of the cells. In this study, we found that inactivated influenza viruses induced changes in the cell surface membrane of Daudi cells resulting in an increased percentage of Con A cap forming cells (30-40%). This phenomenon occurred independently of viral replication and was initiated by adsorption of inactivated viral particles or isolated hemagglutinin and neuraminidase viral glycoproteins. This phenomenon may be due to the binding of Con A molecules to viral receptors and to cell receptors leading to crosslinking of Con A receptors that will induce their mobility and the formation of a cap. Alternatively, experiments performed with cytochalasin B and colchicine suggest that the viral interaction with the cell membrane may have induced changes in the cytoskeleton at the level of microtubules. These changes induced increased lateral movement of the Con A receptors resulting into formation of a cap.

Topics: Adsorption; Cell Membrane; Colchicine; Concanavalin A; Cytochalasin B; Cytoskeleton; Humans; Lymphoma; Orthomyxoviridae; Receptors, Concanavalin A; Tumor Cells, Cultured; Virus Replication

An immunogenic protein with an identical Mr (64 kDa) was isolated from syngeneic concanavalin A-induced lymphoblasts (syn-Con A-blasts) and YAC lymphoma cells, both derived from A mice. The 64-kDa protein was purified by a sequence of biochemical steps: Sephadex G-100 gel filtration, ion-exchange chromatography in a fast protein liquid chromatography system, Con A-Sepharose affinity chromatography, and preparative gel electrophoresis. The immunogenic fraction isolated in each step was moved to the next one, and so on. The immunogenicity of the separated fractions was measured by a lymph node proliferation (LNP) assay, which is indicative of a delayed-type hypersensitivity response. For instance, the final 64-kDa isolated protein of the syn-Con A-blasts induced an efficient LNP response in A mice which was detected after challenge with the final 64-kDa isolated protein of YAC cells. In addition to their identical molecular weight, both proteins were eluted at the same ionic strength and both expressed affinity to Con A-Sepharose beads, suggesting that they were glycosilated. Similar 64-kDa proteins were isolated by a different purification procedure, which was performed in the presence of protease inhibitors, excluding the possibility that the final antigen was an autodigested product. As the 64-kDa protein is immunogenic in the syngeneic host, it may be employed as a immunotherapeutic reagent against the original tumor and perhaps against other tumors expressing the same antigen.

Topics: Animals; Antigens, Differentiation; Chromatography, Affinity; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Female; Hypersensitivity, Delayed; Immunization; Lymphocyte Activation; Lymphocytes; Lymphoma; Mice

An in vitro technique was developed to generate activated rat T cells, with antitumor activity. Splenic mononuclear cells (SMC) from outbred Wistar and inbred Wistar-Munich rats were stimulated with Concanavalin A and recombinant human interleukin-2 (rIL-2) in vitro for 48 h. After 2 days, the nonadherent cells began proliferating and were maintained in rIL-2 for up to 18 days in vitro. FACScan analysis revealed that SMC was a mixture of cell types; however, CD5+ T cells rapidly increased and became the predominant cell type after 5 days in culture. SMC induced cytolysis of YAC-1, but not C6 glioma cells in 4 h 51Cr release assays. In contrast, 5- and 9-day T cells lysed C6 glioma and YAC-1 cells. The C6 cells were admixed with cultured effector cells at various effector-to-target (E:T) ratios and were injected into the right cerebral hemisphere of Wistar and Wistar-Munich rats for a Winn assay. Histopathologic evaluations revealed that a) SMC had no effect; b) 2- and 5-day T cells, injected at E:T ratios greater than 5:1, caused significant reduction in tumor size; and c) 2- or 5-day T cells, at a 40:1 E:T ratio, resulted in little or no histologic evidence of tumor. Eighty-three percent of animals receiving C6 and 5-day mitogen-stimulated lymphokine activated killer cells at an E:T ratio of 40:1 were alive 120 days postinjection (p less than 0.05).

Topics: Animals; Brain Neoplasms; Cell Division; Cell Survival; Cells, Cultured; Concanavalin A; Glioma; Immunophenotyping; Immunotherapy, Adoptive; Interleukin-2; Killer Cells, Lymphokine-Activated; Lymphoma; Male; Neoplasm Transplantation; Rats; Rats, Inbred Strains; Recombinant Proteins; Spleen; Tumor Cells, Cultured

Using five different cell lines as reactive cells, the proliferative inhibitory effect of Con A induced suppressive cells (CISC) was observed. The percentage of inhibition of MLA144 was 93.1%, K562 77.5%, Raji 62.5%, Molt-4 46.4%, and P388 25.9%. It is possible that K562 and MLA144 can be used as reactive cells instead of the peripheral blood mononuclear cells. Supernatants of CISC were also shown to be inhibitory, although less so than CISC. The inhibitory activity of CISC did not seem to be related to prostaglandin. However, its activity could be diminished by anti-T11 monoclonal antibody and complement. These results indicate that CISC is a T11-positive lymphocyte population. The inhibitory activity of CISC from 48 normal donors was determined by this modified method and compared with that of 22 patients with non-Hodgkin's lymphoma. The CISC activity of the patients was significantly lower than that of the normal subjects. Addition of exogeneous IL-2 to the culture enhanced the inhibitory activity in both groups.

Topics: Animals; Antibodies, Monoclonal; Concanavalin A; Humans; Interleukin-2; Leukemia, Myeloid; Lymphoma; T-Lymphocytes, Regulatory; Tumor Cells, Cultured

Thymic-like lymphomas are very sensitive to killing by phytohemagglutinin. To investigate the mechanism of cytotoxicity, we studied the effect of PHA on cytosolic calcium [( Ca2+]i) and cAMP in the S49 mouse lymphoma cell line. PHA produced a slow continuing rise in [Ca2+]i. Estimation of cell number by Coulter counting showed that PHA induced rapid lysis of S49 cells in a dose-dependent manner. Nicardipine (10(-5) M) did not prevent PHA induced cell lysis or [Ca2+]i increase. Also ionomycin (10(-7) M) did not induce cell lysis. The data suggest that PHA induced increase in [Ca2+]i is the result rather than the cause of cell lysis. Elevated intracellular cAMP has an antiproliferative effect on S49 cells. PHA had no effect on cAMP levels in S49 cells. Also S49 cyc- clone which is deficient in Gs was susceptible to killing by PHA. These results suggest that the cytotoxic effect of PHA on S49 cells is rapid, but is not mediated by cAMP generation or an increase in [Ca2+]i, and other mechanisms should be investigated.

Topics: Animals; Calcium; Cell Division; Cell Survival; Concanavalin A; Cyclic AMP; Drug Interactions; Ionomycin; Lymphoma; Mice; Phytohemagglutinins; T-Lymphocytes; Tumor Cells, Cultured

Mouse NK/LY lymphoma cells were modified by Con A or Vibrio cholerae neuraminidase (VCN). Severe damage of the modified cells was not revealed by either supravital staining or light microscopy. Ultrastructural signs of metabolic lowering in the Con-A-treated cells were determined. Some increase in electrophoretic mobility of Con-A-modified cells was found, along with its decrease in VCN-modified cells. The decrease in the pool of proliferating cells was determined in both the types of modification, being more evident in the Con-A-modified material. A marked tumorigenic decrease of Con-A-modified cells was fixed.

Topics: Animals; Cell Division; Concanavalin A; Electrophoresis; Lymphoma; Membrane Potentials; Mice; Mice, Inbred CBA; Neoplasm Transplantation; Neuraminidase; Tumor Cells, Cultured; Vibrio cholerae

A portion of our research involves the investigation of biomarkers as preclinical indicators of toxic stress. Stemming from this effort, we report three unique proteins that phosphorylate and synthesize during the S-phase of lymphoma cells following chemical insult. Mouse lymphoma cell nuclei were physically sorted (using a fluorescence activated cell sorter) from partitions of the cell cycle and specific nuclear proteins in each partition were examined by gel microelectrophoresis. The changes in [32P]-incorporation by stress proteins (SPs) were examined in each of seven partitions following administration of sodium arsenite. Four SPs [80,000-84,000 relative molecular mass (Mr)], designated 'c','b','x', and 'y', underwent significant alterations in [32P]-labeling and each exhibited varying degrees of differential [32P]-incorporation in partition 3 of G1 phase, all five partitions of S-phase, and partition 1 of G2 phase of the cell cycle. Three other predominantly S-phase SPs (designated S1, S2 and S3) were phosphorylated after sodium arsenite treatment. Stress protein S1 was labeled exclusively in S-phase, while proteins S2 and S3 were labeled only in partition 3 of G1 and S-phase. Stress protein S1 possessed an identical isoelectric point, molecular mass, distribution of polypeptide fragments and immunochemical determinants as S-phase SPs found previously in mouse spleen (X') and mouse liver (LP-S). The identical biochemical characteristics of these three S-phase (SPs), found in diverse tissue types, suggest they are homologous.

Topics: Animals; Arsenic; Arsenites; Cell Nucleus; Concanavalin A; Electrophoresis, Gel, Two-Dimensional; Heat-Shock Proteins; Isoelectric Point; Isoproterenol; Liver; Lymphoma; Mice; Molecular Weight; Nuclear Proteins; Peptide Mapping; Phenobarbital; Phosphorus Radioisotopes; S Phase; Sodium Compounds; Spleen; Tumor Cells, Cultured

The method of skin window was used in the investigation of cell reaction of healthy volunteers (25 persons) and patients with malignant melanoma (12 cases), breast cancer (18 cases), and with various malignant lymphomas (6 cases) to a local application of the lectin Concanavalin A (Con A, concentration of 10 mg/ml of saline, 0.02 ml per scarification). The obtained results were compared with cell reaction after the application of saline alone. Correlation between cell reaction and clinical state of patients was studied as well. No significant difference in the amount of lymphocytes after the application of Con A was observed between the group of healthy individuals and the group of patients with malignant tumors. On the contrary, the reaction of neutrophil granulocytes to Con A was significantly lower in cancer patients than in healthy volunteers. After the application of Con A, all preparations showed a higher absolute amount of all cells. Con A induced a significant increase in the amount of lymphocytes in patients after therapy and in poor clinical state. The persons with good clinical state reacted by a slight increase or decrease in the number of lymphocytes to the application of lectin.

Topics: Antibodies, Monoclonal; Breast Neoplasms; Concanavalin A; Humans; Leukocyte Count; Lymphocyte Activation; Lymphocytes; Lymphoma; Melanoma; Mitogens; Skin; Skin Tests

The modifying effect of con A, Vibrio cholerae and non-Vibrio cholerae neuraminidase (VCN, NVCN) on the proliferative potential and immunogenicity of experimental lymphoma NK/Ly cells is shown in vitro and in vivo. The efficacy of immunization by the modified tumour cells of mice with transplantable lymphoma is evaluated. A considerable decrease in the proliferative potential of transplantability and a moderate increase in immunogenicity of Con A treated cells were determined in vitro. The high immunogenicity of VCN-modified cells was found within the limits of the studied immunologic parameters. However, the VCN-MTC immunotherapy had no advantages over Con A-MTC immunotherapy. It is found that the modifying effect of NVCN on a tumour cell is inferior to the effect of Con A and VCN.

Topics: Animals; Concanavalin A; Female; Immunity, Cellular; Immunization; Leukocyte Count; Lymphoma; Mice; Mice, Inbred AKR; Mice, Inbred CBA; Neoplasm Transplantation; Neuraminidase; Rosette Formation; Tumor Cells, Cultured; Vibrio; Vibrio cholerae

Comparisons were made between transformed S49 T-lymphoma cells and normal murine thymocytes in their polyphosphoinositides, inositol polyphosphates and cytosolic free calcium concentrations ([Ca2+]i), and the effects of the T-cell mitogen concanavalin A (Con A) on these properties. 1. The ratios of the polyphosphoinositides to phosphatidylinositol in both exponential-phase S49 cells and mitogen-stimulated thymocytes (G1 phase) were greater than in quiescent (G0-phase) thymocytes. 2. In response to Con A, the amount of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) in S49 cells decreased slightly (17% in 30 min), and this was sufficient to account for the small amounts of inositol phosphates that accumulated. In contrast, it has been shown previously that Con A stimulates a rapid resynthesis of PtdInsP2 in thymocytes and the amounts of inositol phosphates released rapidly exceed the steady-state amount of the PtdInsP2 precursor [Taylor, Metcalfe, Hesketh, Smith & Moore (1984) Nature (London) 312, 462-465]. 3. The [Ca2+]i did not differ significantly in S49 cells and thymocytes before the addition of Con A, and the increases in [Ca2+]i in response to Con A were similar in both types of cell. 4. The [Ca2+]i increase in response to Con A was inhibited by similar concentrations of intracellular cyclic AMP (2-10 microM) in S49 cells and thymocytes, suggesting that similar regulatory mechanisms act on this response in both types of cell. The data demonstrate that the basal [Ca2+]i and phosphoinositide metabolism is similar in both the normal cells and their transformed counterparts. In addition, they suggest that the activated Con A receptors generate very similar signals in the two cell types, and that any perturbations of primary signal transduction to the secondary phosphoinositide and [Ca2+]i responses in the S49 phenotype are quantitative rather than qualitative.

Topics: Adenosine Triphosphate; Aminoquinolines; Animals; Calcium; Cell Line; Concanavalin A; Cyclic AMP; Fluorescent Dyes; Inositol Phosphates; Lymphoma; Mice; Mice, Inbred BALB C; Phosphatidylinositols; T-Lymphocytes

We are interested in understanding the effects of virus infection on lymphocyte function. To approach this question, we used a unique system of two genetically related leporipoxviruses to generate recombinants. One of these viruses, malignant fibroma virus (MV), replicates in many different cell types, including lymphocytes. The other, Shope fibroma virus (SFV), replicates principally in fibroblasts, but cannot replicate in lymphocytes. Fibroblasts infected with SFV received restriction fragments from MV by transfection. Recombinant viruses were selected in vitro for their ability to replicate in lymphocytes. By these means we have identified one restriction fragment, the 10.8-kb BamHI "C" fragment, capable of transferring from MV to SFV the ability to replicate in lymphocytes. A family of recombinants bearing different sized inserts of this restriction fragment has been isolated and is being characterized. Lymphocytotropic recombinants bearing portions of this restriction fragment produce colony morphology in vitro intermediate between MV's plaques and SFV's foci. On the basis of their ability to grow in and suppress mitogen responsiveness of lymphocytes, these recombinants may be classified into four different groups. Group 1 viruses are the most immunosuppressive, whereas those of group 4 are least. These traits correlate with ability to replicate in lymphocytes. Genetic analysis of recombinants indicates that the most immunosuppressive recombinants do not necessarily contain the most fragment C DNA. Therefore, we have identified a restriction fragment, one or more of the genes of which are sufficient to allow an otherwise nonlymphocytotropic virus to replicate in lymphocytes. Additional genetic and immunologic analysis should permit us to determine the structure and function of the protein responsible for this effect.

Topics: Animals; Concanavalin A; DNA Tumor Viruses; DNA, Viral; Fibroblasts; Fibroma Virus, Rabbit; Genes, Viral; Immune Tolerance; Lymphocyte Activation; Lymphocytes; Lymphoma; Polymorphism, Restriction Fragment Length; Poxviridae; Rabbits; Recombination, Genetic; Transfection; Tumor Cells, Cultured; Virus Replication

Topics: Adjuvants, Immunologic; Animals; Concanavalin A; Immunotherapy; Lymphoma; Mice; Mice, Inbred CBA; Neoplasm Transplantation

Topics: Animals; Antibodies, Monoclonal; B-Lymphocytes; Concanavalin A; Female; Killer Cells, Natural; Lymphocyte Activation; Lymphoma; Male; Monkey Diseases; Papio; Precancerous Conditions; T-Lymphocytes; T-Lymphocytes, Regulatory

The effect of interleukin-2 (IL-2) preparations on the susceptibility of YAC cells to mouse natural killer (NK) cells was examined. Crude IL-2 preparations induced a significant decrease in the susceptibility of YAC cells. Highly purified IL-2 preparations, however, did not have a similar effect. Since the IL-2 preparations used in this study were purified from the culture supernatant of Con A activated spleen cells (Con A Sup.), our results indicated the presence in Con A Sup. of a lysis resistance inducing factor (LRIF), distinct from IL-2. We were able to purify LRIF from Con A Sup. by preparative isoelectric focusing and determined its isoelectric pH to be 4.8.

Topics: Animals; Cell Line; Concanavalin A; Cytotoxicity, Immunologic; Interleukin-2; Isoelectric Focusing; Killer Cells, Natural; Lymphokines; Lymphoma; Mice; Mice, Inbred C57BL; Spleen

Two cloned murine helper T lymphocyte (HTL) lines were used to investigate the immunoregulatory properties of highly purified interleukin 2 (IL2). The clone, designated J6.19, secretes lymphokines, including IL2, and proliferates when stimulated with ovalbumin in the presence of I-Ak-bearing spleen cells, while the alloreactive clone, L2, secretes lymphokines, including IL2, and proliferates when stimulated with Mlsa,d-bearing spleen cells. When either clone was exposed to a high concentration of pure IL2 for 1 to 2 days in the absence of either antigen or spleen cells, the HTL became unresponsive to rechallenge with antigen. Unresponsiveness to antigen was indicated by an inability of HTL to proliferate or secrete usual amounts of IL2 or colony-stimulating factor. Within 5-7 days after exposure to IL2, the cloned HTL again responded to antigen. Thus, in addition to being a growth factor for HTL, IL2 can limit the magnitude of the HTL response to antigen. L2 or J6.19 cells could also be induced to secrete lymphokines by the lectin, concanavalin A (Con A) or by a combination of the calcium ionophore, A23187, and phorbol myristate acetate (PMA). After exposure of L2 or J6.19 cells to sufficient IL2 to induce unresponsiveness to antigen, cells were also unresponsive to Con A, as indicated by a reduction in the level of lymphokines secreted. In contrast, lymphokine levels stimulated by A23187/PMA were comparable to those produced by cells not exposed to IL2. The failure of antigen to stimulate lymphokine release and proliferation by HTL previously exposed to IL2 therefore may result from an inability of HTL to recognize antigen or to transduce effectively the antigen recognition signal. Several T cell surface molecules are known to be involved in antigen activation of HTL; these include the antigen receptor and the "associative recognition" structures L3T4 and LFA-1. We observed that L2 cells, rendered unresponsive to antigen by exposure to IL2, expressed normal levels of antigen receptor, as identified by the monoclonal antibody, KJ16-133.18. Furthermore, expression of L3T4 and LFA-1 was not decreased. Unresponsiveness to antigen induced by IL2 thus could not be correlated with decreases in the expression of antigen receptors, L3T4, or LFA-1. Unresponsive HTL may therefore be capable of recognizing antigen but the signal generated by antigen binding may be attenuated during its transduction, resulting in the failure of cloned HTL to proliferate or secrete ly

Topics: Animals; Antigens; Calcimycin; Cell Line; Concanavalin A; Interleukin-2; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred CBA; Receptors, Antigen, T-Cell; Recombinant Proteins; T-Lymphocytes, Helper-Inducer; Tetradecanoylphorbol Acetate

Thymocytes of leukemic AKR mice were analysed with a fluorescence-activated cell sorter (FACS) using monoclonal antibodies (MAbs) reactive with H-2Kk and H-2Dk and conventional hetero-antibodies against MuLV gp-70 antigens. Comparison was made with thymocytes of the low-leukemia H-2k strain C3H, as well as with thymocytes of young AKR mice, late preleukemic AKR mice (6-9 months old) and mitogen-stimulated thymocytes of young AKR mice. Expression of H-2 antigens increased on cells of the majority of tumors, on thymocytes of some late preleukemic mice and on mitogen-stimulated thymocytes. An increase in H-2K and H-2D antigens was noted: imbalance of the K/D ratio in favor of H-2D was observed mostly in tumors with relatively lower total amounts of H-2K and D antigens; ratio shifts in favor of H-2K were also found, mostly in tumors with relatively higher amounts of H-2K and D antigens. Increased expression of MuLV antigens was found on cells of all tumors and on thymocytes of several late preleukemic mice. We show that in primary spontaneous AKR leukemias, in spite of large individual differences, the expression of high amounts of MuLV gp70 is not random, but is associated with high expression of H-2K and H-2D antigens. The same phenomenon is seen in thymocytes of preleukemic mice, but in mitogen-stimulated thymus cells increase of H-2 expression is not accompanied by increase of MuLV gp70.

Topics: Animals; Antigens, Viral; Concanavalin A; H-2 Antigens; Histocompatibility Antigen H-2D; Leukemia Virus, Murine; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred C3H; Phenotype; T-Lymphocytes

The specific activity in cells from lymph nodes, spleen and thymus was 32, 28 and 25 nmol/min per mg of cytosol protein, respectively, whereas that in bone marrow cells was significantly lower (10 units/mg of protein). No difference in specific DAN activity between isolated B- and T-lymphocytes was observed. Two types of lymphoid mouse cell lines (MOPC-31C plasmacytoma cells, S49 Cyc- lymphoma cells) showed specific activities similar to the normal lymphoid cells. In concanavalin A- stimulated spleen lymphocytes in culture there was a rapid increase in DAN activity shortly after maximum DNA synthesis, reaching a plateau 2-3 times the original level. The enzyme (DAN) of mouse tissues possessed the characteristic properties previously detected for the rat enzyme.

Topics: Animals; Bone Marrow; Cell Line; Concanavalin A; Cytosol; Kinetics; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Lymphoma; Male; Mice; Mice, Inbred Strains; Nucleotidases; Organ Specificity; Plasmacytoma; Spleen; Thymus Gland

We have studied early transmembrane events in mouse alloimmune cytotoxic T-lymphocyte (LC7, H-2b) activation by specific target cells (mouse mastocytoma P815, H-2d) and a mitogenic lectin, Con A, by using stopped-flow fluorometry with three different fluorescent probes. After binding to target cells (P815), cytotoxic T lymphocytes (LC7) first increased their membrane fluidity and, then, calcium was released from intracellular stores. After that, there was a calcium influx from the external medium into the T lymphocytes. This calcium influx was blocked by calcium antagonists (verapamil or diltiazem). The same sequence of events was also observed in the activation of T lymphocytes (LC7) by Con A and in the response of specific target cells (P815) after cytotoxic T lymphocytes (LC7) binding. Nonspecific (syngeneic) target cells (mouse lymphoma EL-4, H-2b) did not cause any early transmembrane events in cytotoxic T lymphocytes (LC7, H-2b).

Topics: Animals; Biological Transport; Calcium; Cell Line; Concanavalin A; Cytotoxicity Tests, Immunologic; Diltiazem; Fluorescent Dyes; Fluorometry; Lymphocyte Activation; Lymphoma; Mast-Cell Sarcoma; Membrane Fluidity; Mice; T-Lymphocytes, Cytotoxic; Verapamil

Sendai virus vesicles were used as vehicles for the insertion of various cell membranes into different cell lines. The transplantation efficiency was measured by using FITC-labelled concanavalin A (Con A) or monoclonal antibodies against the T-cell marker Lyt 2 and the major histocompatibility complex product H-2Db in a fluorescence-activated cell sorter. Results indicate that it is possible to transplant mitogen responsiveness to certain cell types. Con A stimulation of T-cell membrane transplanted BW 5147 showed that it is possible to induce a mitogen dose-dependent T-cell growth factor production. Consequently this method appears to be an attractive model for further study of the properties of the membrane structures involved in mitogen triggering of cells.

Topics: Animals; Cell Fusion; Cell Line; Cell Membrane; Concanavalin A; Dose-Response Relationship, Immunologic; Interleukin-2; Leukemia L1210; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred AKR; Mice, Inbred C57BL; Mice, Inbred DBA; Receptors, Antigen, T-Cell; Receptors, Concanavalin A; T-Lymphocytes; Thymus Neoplasms

Considerable evidence documents the importance of co-factors, including the immune response, in expression of oncogenicity of tumour viruses. To determine whether a common protozoal infection that can depress lymphocyte function alters manifestations of oncogenic virus infection, a mouse model of Toxoplasma infection with depressed T lymphocyte function was developed. In this model, Toxoplasma depressed blastogenic transformation to the T-cell mitogen Concanavalin A and primary antibody response to sheep red blood cells which requires T cell help. Uninfected and Toxoplasma-infected mice were then infected with Moloney leukaemia or Moloney sarcoma viruses and development of lymphoma and sarcoma were evaluated. Toxoplasma infection, which induced depression of T-cell function, decreased the incidence of Moloney sarcoma virus induced rhabdomyosarcomas but did not alter progression or regression of tumour in those mice that developed tumour. Conjoint infection with Toxoplasma and Moloney leukaemia virus did not increase incidence of lymphoma when compared with incidence of lymphoma in mice infected with Moloney leukaemia virus alone.

Topics: Animals; Antibody Formation; Concanavalin A; Female; Leukemia, Experimental; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred BALB C; Moloney murine leukemia virus; Moloney murine sarcoma virus; Rhabdomyosarcoma; Sarcoma, Experimental; T-Lymphocytes; T-Lymphocytes, Helper-Inducer; Thymus Gland; Toxoplasmosis

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

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

The ontogeny of proliferative and cytotoxic responses to concanavalin A (Con A) and interleukin 2 (IL 2) in C57BL/6J (B6) fetal thymus (FT) was investigated. Embryonic thymocytes were either taken from embryos at different times of gestation or from 14 day B6 FT that were maintained as organ cultures for various times. It was found that the B6 FT could proliferate to Con A and EL4 SN (an IL 2 containing culture supernatant) in a synergistic fashion. This synergy between Con A and EL4 SN was first observed at the 16th to 17th day of gestation. A similar differentiation process took place in 14-day FT that had been maintained as organ cultures; the synergy between Con A and EL4 SN was first observed after 3 days in organ culture. This synergy increased with increasing time of organ culture, and was most evident after 10 days. The synergy between Con A and EL4 SN was also observed when the EL4 SN was replaced with IL 2 which had been purified from crude EL4 SN to apparent homogeneity. B6 FT could also form cytotoxic T lymphocytes (CTL) on stimulation with Con A and EL4 SN. Con A-activated CTL (polyspecific) were detected by including phytohemagglutinin in the assay medium. CTL response was first detected in the 17-day fetal thymus by using this assay. In organ cultures, CTL responses were first detected after 4 days in organ culture, and reached peak levels after 12 to 14 days. The CTL precursor (CTL-P) frequencies in the B6 FT after 2, 5, 10, and 14 days in organ culture were less than 1/10,000, 1/2232, 1/297, and 1/70, respectively; the corresponding CTL-P frequency in adult thymus was 1/60. After 6 days in organ culture, B6 FT could also form CTL in response to Con A and pure IL 2. This finding suggests that the ability to synthesize other differentiation factors that are required for CTL responses is acquired at an early time of thymic differentiation.

Topics: Animals; Cell Differentiation; Cell Line; Concanavalin A; Cytotoxicity, Immunologic; Female; Interleukin-2; Lymphocyte Activation; Lymphoma; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Organ Culture Techniques; Pregnancy; Stem Cells; T-Lymphocytes, Cytotoxic; Tetradecanoylphorbol Acetate; Thymus Gland

Ultrastructural studies of normal and neoplastic lymphocytes are presented that qualitatively and quantitatively assess the central cell organelle currently used by surgical pathologists in the classification of non-Hodgkin's lymphoma, the nucleus. Events occurring during normal lymphocyte transformation can be used to appreciate essential mechanisms involved in producing the appearance of the nucleus as seen by microscopy. Quantitation of nuclear subcompartments by morphometric image analysis reveals that determination of nuclear size is primarily due to the ribonucleoprotein materials distributed between condensed chromatin masses, the interchromatinic (euchromatin or nuclear matrix) region. Furthermore, such investigations show that amounts and distribution of condensed chromatin in lymphocyte nuclei cannot be adequately assessed from routine histologic sections. Ultrastructural morphometric analysis of representative cases of the principal subtypes of NHL indicates that the atypical morphologic appearance of neoplastic lymphocytes results from a complex interplay between total amounts of condensed chromatin in nuclei and the size of individual aggregates of condensed chromatin, one or both of which may be abnormal in NHL. Abnormalities of interchromatinic materials are also likely involved in ordering the gross appearance of the nucleus. Understanding of both the dynamic capabilities of the nucleus, and the organization of and interplay between the various subcompartments of this organelle will be helpful in improving the classification of NHL by surgical pathologists.

Topics: Animals; B-Lymphocytes; Biopsy; Cell Nucleus; Chromatin; Concanavalin A; Escherichia coli; Humans; Lipopolysaccharides; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Lymphoma; Mice; T-Lymphocytes

It is reported that concanavalin A (conA) and wheat germ agglutinin (WGA) have a differential binding pattern on normal mouse spleen lymphocytes and the surface of Dalton's lymphoma cells. It is suggested that sialic acid on the cell surface controls the expression of lectin binding sites. Further, it has been observed that the increased release of sialic acid from cell surfaces after cis-dichlorodiammine platinum (II) (cis-Platin) treatment is due to the increased activity of sialidase.

Topics: Animals; Binding Sites; Cisplatin; Concanavalin A; Fluorescein-5-isothiocyanate; Fluoresceins; Lectins; Lymphocytes; Lymphoma; Mice; Mice, Inbred DBA; Microscopy, Fluorescence; N-Acetylneuraminic Acid; Neuraminidase; Sialic Acids; Thiocyanates; Wheat Germ Agglutinins

The effect of castanospermine on the processing of N-linked oligosaccharides was examined in the parent mouse lymphoma cell line and in a mutant cell line that lacks glucosidase II. When the parent cell line was grown in the presence of castanospermine at 100 micrograms/ml, glucose-containing high-mannose oligosaccharides were obtained that were not found in the absence of inhibitor. These oligosaccharides bound tightly to concanavalin A-Sepharose and were eluted in the same position as oligosaccharides from the mutant cells grown in the absence or presence of the alkaloid. The castanospermine-induced oligosaccharides were characterized by gel filtration on Bio-Gel P-4, by h.p.l.c. analysis, by enzymic digestions and by methylation analysis of [3H]mannose-labelled and [3H]galactose-labelled oligosaccharides. The major oligosaccharide released by endoglucosaminidase H in either parent or mutant cells grown in castanospermine was a Glc3Man7GlcNAc, with smaller amounts of Glc3Man8GlcNAc and Glc3Man9GlcNAc. On the other hand, in the absence of castanospermine the mutant produces mostly Glc2Man7GlcNAc. In addition to the above oligosaccharides, castanospermine stimulated the formation of an endoglucosaminidase H-resistant oligosaccharide in both cell lines. This oligosaccharide was characterized as a Glc2Man5GlcNAc2 (i.e., Glc(1,2)Glc(1,3)Man(1,2)Man(1,2)Man(1,3)[Man(1,6)]Man-GlcNAc-GlcNAc). Castanospermine was tested directly on glucosidase I and glucosidase II in lymphoma cell extracts by using [Glc-3H]Glc3Man9GlcNAc and [Glc-3H]Glc2Man9GlcNAc as substrates. Castanospermine was a potent inhibitor of both activities, but glucosidase I appeared to be more sensitive to inhibition.

Topics: Acetylglucosaminidase; Alkaloids; Animals; Cell Line; Chromatography, Gel; Chromatography, High Pressure Liquid; Concanavalin A; Glucosidases; Glycoproteins; Indolizines; Lymphoma; Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; Methylation; Mice; Oligosaccharides

A monoclonal antibody, AN-18.17.24, specific for murine interferon-gamma (IFN-gamma) was produced by immunizing Wistar rats with IFN-gamma secreted by a T-cell lymphoma, L12-R4, upon stimulation with phorbol myristic acetate (PMA). Antiviral activity as well as tumoricidal activation induced by PMA-stimulated L12-R4 cell supernatant or by Con A-stimulated normal spleen cells were neutralized at the same extent by AN-18 monoclonal antibody. Moreover, depletion experiments showed that inhibition of tumoricidal macrophage activation must be ascribed to the direct binding of the IFN-gamma molecule by AN-18 MAb and not to the interference of the monoclonal antibody with the cell surface IFN-gamma receptor. These studies conclusively demonstrate that in supernatants of T lymphocytes stimulated with polyclonal activators IFN-gamma was the only molecule responsible for macrophage activation in tumor cell killing.

Topics: Animals; Antibodies, Monoclonal; Cell Line; Concanavalin A; Cytotoxicity, Immunologic; Female; Interferon-gamma; Lymphoma; Macrophage Activation; Mast-Cell Sarcoma; Mice; Rats; T-Lymphocytes; Tetradecanoylphorbol Acetate

Early transmembrane events of tumour cells (mouse myeloma X5563 and lymphoma RDM4) after binding of a monoclonal antibody against mouse MHC antigen and a mitogenic lectin, Con A, were examined by stopped-flow fluorometry with 3 different fluorescent probes. The results showed that membrane fluidities of the cells increased first after binding of anti H-2Kk monoclonal antibody (11-4.1), then calcium was released from intracellular stores into the cytoplasma, and lastly calcium influx occurred from the external medium into the cytoplasma. While Con A only induced calcium influx from the external medium into the cytoplasma.

Topics: Animals; Antibodies, Monoclonal; Biological Transport, Active; Calcium; Cell Line; Cell Membrane; Concanavalin A; Cytoplasm; H-2 Antigens; Lymphoma; Membrane Fluidity; Mice; Multiple Myeloma; Neoplasms, Experimental; Spectrometry, Fluorescence

Sixty-three patients with Hodgkin's disease (HD) or non-Hodgkin's lymphoma (NHL) were studied to analyze the mechanisms responsible for impaired in vitro lymphocyte reactivities to the mitogen concanavalin A. Lymphocytes from 43 of the 52 untreated patients acquired enhanced in vitro responsiveness after preculturing in media alone for 3 days. However, 38 of the untreated patients failed to achieve entirely normal lymphocyte responses after preculturing . Suppressor cells were detected in 25 patients, but the intensity of suppression was much less than expected when compared with the severity of in vitro impairments. Suppressor activity did correlate with certain clinical characteristics in NHL, whereas no correlation was observed for HD. In contrast to the untreated patients, successfully treated patients demonstrated either normal responses or profound, irreversible impairments. The data indicate that several mechanisms which usually coexist can contribute to the impaired in vitro lymphocyte responses in untreated HD and NHL, and that a single, irreversible type of mechanism explains the impaired reactivities in successfully treated patients.

Topics: Adult; Cell Division; Cells, Cultured; Concanavalin A; Female; Hodgkin Disease; Humans; Lymphocyte Culture Test, Mixed; Lymphoma; Male; Middle Aged; Monocytes; T-Lymphocytes, Regulatory

The cultured murine B cell lymphoma, 70Z /3, can be induced to express membrane IgM ( mIgM ) after exposure to lipopolysaccharide (LPS) or T cell-derived factors. The kinetics and magnitude of the responses have been compared in wild type 70Z /3 cells and three variants by using flow cytometric analysis and immunoprecipitation. Wild type 70Z /3 cells respond to LPS more quickly and with twofold greater mIgM than to concanavalin A-induced spleen cell supernatant (CAS). Variants were selected for their abnormal mIgM expression in response to LPS, but individual variants also showed normal, aberrant, or no response to CAS. When cells were induced with suboptimal amounts of LPS and CAS, a synergistic effect on the magnitude of mIgM expression was seen in wild type and variant cells. This suggests that both inducing agents are utilizing some part of a common inductive mechanism. The different responses of the variant cell lines will allow further genetic dissection and comparison of the mIgM expression pathways used in response to LPS and CAS.

Topics: Animals; B-Lymphocytes; Cell Line; Concanavalin A; Drug Synergism; Immunoglobulin Light Chains; Immunoglobulin M; Immunoglobulin mu-Chains; Lipopolysaccharides; Lymphocyte Activation; Lymphokines; Lymphoma; Male; Mice; Phenotype; Rats; Rats, Inbred Strains; Receptors, Antigen, B-Cell; T-Lymphocytes

Monoclonal antibodies against murine immune interferon (IFN-gamma) were produced by fusing the murine nonsecreting myeloma cell line P3.X63.Ag8.653 with spleen cells from rats immunized with IFN-gamma-containing supernatants obtained by stimulating a T-cell lymphoma, L12-R4, with phorbol 12-myristate 13-acetate. Supernatants from a twice-cloned hybridoma (AN-18.17.24) were found to neutralize and to adsorb in depletion experiments up to 27 units of mouse IFN-gamma but not equivalent amounts of mouse leukocyte or fibroblast IFNs. The AN-18.17.24 monoclonal antibody neutralized to the same extent mouse IFN-gamma from different sources--namely, (i) concanavalin A-stimulated spleen cells, (ii) alloantigen-stimulated spleen cells, and (iii) monkey fibroblasts transfected with the cloned gene of murine IFN-gamma. Moreover, the monoclonal antibody displayed species specificity, since it did not neutralize IFN-gamma of human origin. Binding inhibition experiments with murine IFN-gamma preparations exposed to enzymatic or physicochemical degradation demonstrated that the protein moiety and not the carbohydrate residues were responsible for the binding to the AN-18.17.24 monoclonal antibody. Finally, this monoclonal antibody immunoprecipitated two molecular species of IFN-gamma of about 16.8 and 17.8 kilodaltons, respectively, from [35S]methionine- or [3H]glucosamine-labeled supernatants of stimulated L12-R4 cells.

Topics: Animals; Antibodies, Monoclonal; Cell Line; Concanavalin A; Haplorhini; Interferon-gamma; Isoantigens; Lymphoma; Mice; Mice, Inbred C57BL; Molecular Weight; Rats; Spleen; Tetradecanoylphorbol Acetate

We have examined a concanavalin A-resistant (Con AR) Chinese hamster ovary cell (CR-7) that has a defect in the synthesis of asparagine-linked oligosaccharides and consequently an altered expression of membrane carbohydrate. The CR-7 mutant, which has a decreased ability to incorporate mannose into oligolipid and membrane glycoprotein and an increased membrane fucose, was more sensitive to natural killer (NK) cell lysis than the parental wild type (CHO-WT). Splenocytes mediating the lysis of the CR-7 line were asialo GM1+, nonadherent, IFN stimulatable, absent in the bg/bg mutant, and co-fractionated on Percoll density gradients with cells mediating lysis of the YAC-1 murine lymphoma. The increase in NK lysis correlated with enhanced binding of NK cells to the mutant determined by adsorption on tumor monolayers, cold target inhibition, and target binding analysis. A revertant of CR-7 (RCR-7), which showed wild-type levels of NK lysis, was intermediate in its ability to bind or cold target inhibit NK cells. The CR-7, CHO-WT, and RCR-7 lines were equally sensitive to hypotonic lysis and cytotoxicity by human lymphokine-activated killer (LAK) cells suggesting that the mutation did not nonspecifically alter membrane fragility. The NK-sensitive CR-7 line was less tumorigenic after subcutaneous injection in nude mice when compared with the parental CHO-WT or RCR-7 lines. This decreased tumorigenicity could be reversed by the i.v. injection of antiserum directed at the NK cell determinant asialo GM1. In conclusion, a ConAR tumor cell with a demonstrable oligosaccharide biosynthetic defect, exhibited enhanced NK lytic sensitivity and was poorly tumorigenic in vivo, a feature which may also be a consequence of its altered NK reactivity.

Topics: Absorption; Animals; Binding, Competitive; Cell Communication; Cell Line; Cell Membrane; Concanavalin A; Cricetinae; Cricetulus; Cytotoxicity, Immunologic; Killer Cells, Natural; Lymphoma; Male; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Mice, Nude; Neoplasm Transplantation; Oligosaccharides

Phytohemagglutinin (PHA) and concanavalin A (Con A) were used as probes to detect changes in the cell surface of Dalton's lymphoma, sarcoma-180 and Ehrlich's carcinoma after short in vitro exposure to acriflavine. Dye-treated cells showed enhancement of agglutination both by PHA and Con A, and such enhancement was found to be dependent on the time of exposure and concentration of acriflavine. However, PHA-induced percent agglutination seemed to be much higher than that of Con A among the 3 cell types. There were also marked differences among the 3 cell types in order of their sensitivity to lectin-mediated agglutination. The strength of the response was greater in lymphoma to both PHA and Con A than that of sarcoma-180 and carcinoma cells, which appeared to be most resistant. Acriflavine, which is known as an intercalative agent with DNA, induces cell surface changes by promoting lectin-mediated cellular agglutination.

Topics: Acriflavine; Agglutination; Aminoacridines; Animals; Carcinoma, Ehrlich Tumor; Cell Membrane; Cells, Cultured; Concanavalin A; Lymphoma; Male; Mice; Phytohemagglutinins; Sarcoma 180

Concanavalin agglutinin (Con A) binding sites were studied in paraffin embedded lymph node specimens of reactive follicular hyperplasia (12 cases) and follicular lymphoma (37) using the avidin-biotin-peroxidase complex method, and the results were compared with those of Peanut agglutinin (PNA) and lysozyme stains. Very similar to the PNA stain, two categories of Con A receptor sites were observed: cytoplasmic and cell surface. In the reactive lymph nodes, the cells showing cytoplasmic receptor sites (CR+ cells) corresponded to macrophage-histiocytes and possibly dendritic reticulum cells in the H & E stained sections, while those showing cell surface receptor sites (SR+) corresponded to lymphoid cells. Unlike the PNA binding, however, the staining reaction of SR+ lymphoid cells was weak, and another staining pattern, a dot-like stain, was observed in some lymphocytes, both SR+ and SR-. In follicular lymphomas, CR+ histiocytes were distinctly displayed within the follicular centers in 25 of 37 cases, including 12 cases in which PNA stains on adjacent or nearby sections were negative for intrafollicular macrophage-histiocytes. Similarly, Con A stains were positive for the intrafollicular CR+ cells in four of the five cases in which lysozyme stains were negative. Many of these intrafollicular CR+ cells contained inclusion-like cytoplasmic globules and/or vacuoles, a hallmark of the large CR+ cells of germinal centers. These observations suggest that macrophage-histiocytes of presumably germinal center origin are retained in neoplastic follicular centers in varying degrees, and Con A might be a useful marker for macrophage-histiocytes in paraffin-embedded routine pathological specimens, in addition to the currently accepted markers, PNA and lysozyme.

Topics: Cell Membrane; Concanavalin A; Cytoplasm; Female; Histiocytes; Histocytochemistry; Humans; Hyperplasia; Lymph Nodes; Lymphoma; Male; Middle Aged; Receptors, Concanavalin A; Staining and Labeling

Tumor pairs, selected on the basis of their different capacities to metastasize in vivo (SP73/AS and ASML from the rat, Eb/ESb from the mouse), have been assayed for their membrane associated actin through the DNase inhibition assay. It is found that, provided inhibitions per cell are corrected for the influence of gross heterogeneities in size distributions, the more metastatic tumor cells have significantly higher DNase I inhibitions than their less invasive counterparts. This observation, which extends our previous study of normal recirculating lymphocytes, is rationalized by postulating a participation of these actin pools to a property critical for both normal recirculation and metastatic spreading, arguments are presented which favor cell surface deformability as a possible candidate.

Topics: Actins; Adenocarcinoma; Animals; Concanavalin A; Deoxyribonuclease I; Endodeoxyribonucleases; Female; Fibrosarcoma; Lymphoma; Male; Neoplasm Metastasis; Neoplasms; Rats

Monocyte depleted blood lymphocytes were analyzed in 28 patients with non-Hodgkin's lymphoma in early unmaintained complete clinical remission, in 8 long-term complete remission patients, and in 31 age-matched healthy controls, by SRBC rosetting and DNA synthesis induced by concanavalin A, pokeweed mitogen and PPD antigen. The median observation time from testing was 22 months (range 10-49 months). Early complete remission patients with a normal mitogen induced lymphocyte stimulation remained free of relapses. In contrast, patients who relapsed during the observation period had lymphocytopenia (p less than 0.05), reduced lymphocyte responses to mitogen (p less than 0.001) and to PPD antigen (p less than 0.05) as compared with healthy controls. Long-term complete remission patients did not differ from healthy controls with respect to lymphocyte counts and response to mitogen stimulation.

Topics: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Concanavalin A; DNA; Female; Follow-Up Studies; Humans; Lymphocyte Activation; Lymphocytes; Lymphoma; Male; Middle Aged; Mitogens; Pokeweed Mitogens; Prognosis; Recurrence; Rosette Formation; Tuberculin

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

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

It has been demonstrated that the human histiocytic lymphoma-derived cell line U937, which has monocytoid characteristics, responds to a concanavalin A-induced T-cell-derived suppressor supernatant (T-SFS) with the release of a factor markedly suppressing mitogen-stimulated proliferation of normal peripheral blood lymphocytes. The suppressor material is not dialyzable, appears within 2 hr of exposure of U937 cells to the T-SFS, persists for at least 24 hr, and has a Mr of approximately 40,000 by gel chromatography. The suppressor factor does not affect the proliferation of continuous T- and B-lymphoid cell lines, distinguishing it from the inhibitor of DNA synthesis also released by U937, but appears to be specific for a stage of activation of normal lymphocytes that is independent of (a) utilization of interleukin-2 and (b) inhibition of production of interleukin-2.

Topics: B-Lymphocytes; Cell Line; Concanavalin A; Humans; Interleukin-2; Lymphocyte Activation; Lymphokines; Lymphoma; Macrophages; Molecular Weight; Pokeweed Mitogens; Suppressor Factors, Immunologic; T-Lymphocytes

Rat polymorphonuclear leukocytes (PMN) pretreated with supernatant of a spleen cell culture stimulated with concanavalin A (Con A sup) inhibited 3H-thymidine uptake of various syngeneic, allogeneic, and xenogeneic tumor cells. Con A sup-treated PMN were cytostatic not only to tumor cells but also to embryonal fibroblasts and normal Con A blasts. Stimulated PMN also killed some tumor cells, as shown by 3H-uridine release assay. Cytostasis by Con A sup-treated PMN began at a very early phase of incubation, but in the cytolysis assay, incubation for more than 12 hr was required to obtain significant killing. The time required for activation of cytostatic PMN by Con A sup was short (5 min was enough) if high titers of Con A sup were used for activation. PMN-stimulating activity of Con A sup was not abrogated by treatment at 60 degrees C for 30 min or by change of pH from 1 to 11. The mechanisms of cytotoxicity by Con A sup-treated PMN are discussed.

Topics: Animals; Cell Survival; Cells, Cultured; Concanavalin A; Cytotoxicity, Immunologic; Drug Stability; Lymphokines; Lymphoma; Macrophages; Mice; Neutrophils; Rats; Rats, Inbred F344; Rats, Inbred Lew; Rats, Inbred Strains; Spleen; Time Factors

A tetrazolium salt has been used to develop a quantitative colorimetric assay for mammalian cell survival and proliferation. The assay detects living, but not dead cells and the signal generated is dependent on the degree of activation of the cells. This method can therefore be used to measure cytotoxicity, proliferation or activation. The results can be read on a multiwell scanning spectrophotometer (ELISA reader) and show a high degree of precision. No washing steps are used in the assay. The main advantages of the colorimetric assay are its rapidity and precision, and the lack of any radioisotope. We have used the assay to measure proliferative lymphokines, mitogen stimulations and complement-mediated lysis.

Topics: Animals; Cell Line; Cell Survival; Colorimetry; Concanavalin A; Cytotoxicity Tests, Immunologic; Dose-Response Relationship, Immunologic; Interleukin-2; Lipopolysaccharides; Lymphocyte Activation; Lymphocytes; Lymphoma; Macrophage Activation; Mice; Mice, Inbred BALB C; Tetrazolium Salts; Thiazoles

YAC-1 lymphoma cells, both when cultured in vitro and when passaged in ascites form in vivo, synthesize gangliosides (means of 22.1 and 14.7 nmol lipid-bound sialic acid isolated per 10(8) cells, respectively) with potent inhibitory effects on mitogen- and antigen-induced lymphoproliferation: 10 to 30 nmol highly purified YAC-1 gangliosides/ml caused greater than 90% inhibition of proliferative responses of murine lymphocytes to concanavalin A, lysozyme (a soluble specific antigen), and allogeneic cells (mixed-lymphocyte response). Measureable quantities of these gangliosides were shed by the tumor cells in vitro and also were recovered from the ascites fluid in vivo. Furthermore, the gangliosides isolated from ascites fluid (mean of 15.3 nmol/ml) had inhibitory activity of a magnitude similar to that of the gangliosides isolated from the tumor cells. Therefore, significant inhibition of normal lymphoproliferative responses by tumor-derived gangliosides occurred at ganglioside concentrations which are actually present in the fluid surrounding the tumor cells in vivo. These results support the hypothesis that shedding of gangliosides may serve to protect tumor cells from host immune destruction.

Topics: Cells, Cultured; Concanavalin A; Gangliosides; Humans; Lymphocyte Activation; Lymphoma; Muramidase

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

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

Various cloned murine T cell hybridomas and T cell lymphomas were evaluated for their ability to produce interferon (IFN). Two T cell tumor clones, L12-R1 and L12-R4, derived from the spontaneously in vitro transformed cell lines L12 originally established from fetal calf serum-primed C57BL/6 spleen cells were found to produce high IFN amounts upon mitogen stimulation. Phorbol myristate acetate led to maximal IFN production (2187 IU) by L12-R4 cells at concentrations of 2 x 10(-7) M, whereas concanavalin A and phytohemagglutinin induced lower levels of IFN synthesis (160 to 243 IU). None of the cell lines tested produced IFN constitutively or upon lipopolysaccharides stimulation. The IFN was characterized as immune (y) by being labile at pH 2 and neutralized by two rabbit anti-murine IFN-y antisera but not by antiserum to murine leukocyte (alpha) and fibroblast (beta) IFN. Phenotypic characterization of IFN-y-producing cells showed the L12 clones to be Thy-1.2+, Lyt-1+, 23-, and Ig-. The L12-R4 tumor cell therefore provide a unique source of IFN-y for purification, and may represent a useful model for studying the molecular mechanisms involved in T cell differentiation leading to IFN-y production.

Topics: Animals; Concanavalin A; Fibroblasts; Humans; Hybridomas; Interferons; Lymphoma; Mice; Mice, Inbred C57BL; Neutralization Tests; Phytohemagglutinins; Species Specificity; T-Lymphocytes; Tetradecanoylphorbol Acetate

Topics: Animals; B-Lymphocytes; Cell Transformation, Neoplastic; Concanavalin A; Epitopes; Female; Histocompatibility Antigens Class II; Immunoglobulin M; Interleukin-1; Lipopolysaccharides; Lymphokines; Lymphoma; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Proteins; Receptors, Antigen, B-Cell; Time Factors

When stained for reactive sialyl groups with fluorescein-labelled Aprotinin (FLA), lymphocytes of three diffuse lymphomas were uniformly faintly fluorescent. The nodules of a nodular lymphocytic lymphoma showed dimly fluorescing lymphocytes surrounded by brightly fluorescing, apparently normal cells. The spleens of eight patients with Hodgkin's disease showed involvement in six cases. With FLA, the two uninvolved spleens contained only brightly fluorescing lymphocytes, whereas the foci of Hodgkin's lesions in the six spleens and in eight involved lymph nodes from a further eight patients contained varying proportions of brightly and dimly fluorescing lymphoid cells. Mononuclear Hodgkin's cells and bi- or multinucleated Reed-Sternberg cells fluoresced faintly. Fluorescein-labelled Ricinus communis agglutinin (FL-RCA) for galactose, and Concanavalin A (FL-Con A) for mannose or glucose, showed eosinophils, reticulin and collagen fibres especially in nodular sclerosing Hodgkin's disease, whereas all lymphocytes, Hodgkin and Reed-Sternberg cells stained faintly with either lectin. The reduction of reactive sialyl groups in malignant lymphocytes of lymphomas and Hodgkin's lesions is similar to that in lymphocytic leukaemias. It is suggested that in Hodgkin's disease these lymphocytes together with the Hodgkin and Reed-Sternberg cells represent the malignant component, whereas the brightly fluorescent normal lymphocytes, together with histiocytes, eosinophils (and neutrophils) represent a reactive component in the lesions. Similarly, the reactive lymphocytes in sarcoid lesions and sinus histiocytosis were brightly fluorescing.

Topics: Aprotinin; Bone Marrow; Concanavalin A; Fluorescent Antibody Technique; Hodgkin Disease; Humans; Lectins; Lymph Nodes; Lymphocytes; Lymphoma; Plant Lectins; Sialic Acids; Spleen; Staining and Labeling

Untreated patients with Hodgkin's disease are known to have significant impairment of cellular immunity. Recent studies have demonstrated that effector T cells from these patients have increased sensitivity to the suppression mediated by two normal immunoregulatory cells, ie, suppressor monocytes and suppressor T cells. Thus, increased sensitivity to suppression may be a common cause of multiple abnormalities of cellular immunity. Patients achieving long-term disease-free survival after chemotherapy have also been studied. Although they are no longer anergic, they have persistent reductions in peripheral blood E rosettes and T cell proliferation. Increased sensitivity to suppressor monocytes and T cells also persists. These abnormalities do not appear to be caused by the treatment since they were not detected in diffuse histiocytic lymphoma patients surviving after similar chemotherapy. Immunologic studies in family members are required to determine whether these abnormalities are a permanent immunologic deficit acquired only with the development of Hodgkin's disease or an inherited characteristic that predisposes a patient to develop Hodgkin's disease.

Topics: Concanavalin A; Hodgkin Disease; Humans; Immunity, Cellular; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphoma; T-Lymphocytes; T-Lymphocytes, Regulatory

By fusing primed murine lymphocytes with a syngeneic T cell lymphoma, we have been able to select for H-2-restricted, virus-specific cytotoxic T cell hybridomas (CTH). These T cell hybrids, which replicate in ordinary tissue culture medium or in ascites, are capable of lysing virally infected target cells, and their activity is facilitated by the presence of lectins in the assay medium. Unlike cells mediating lectin nonspecific lysis, these hybridomas are H-2 restricted and specific for single viral proteins. The ability to maintain these cells in culture for over 18 mo and to pass them in vivo without loss of activity or specificity indicates that they will provide sufficient material for the analysis of surface proteins and genetic information required for the recognition and lysis of virally infected cells by killer T cells.

Topics: Animals; Concanavalin A; Cytotoxicity, Immunologic; Female; H-2 Antigens; Hybridomas; Lymphoma; Mice; Mice, Inbred C3H; Reoviridae Infections; T-Lymphocytes

Human plasma contains chemoattractant activity for cultured cells from the mouse thymic lymphoma 6C3HED and also for lymphoblasts from concanavalin A-stimulated mouse spleen cells. A major portion of the attractant activity for both cell types could be attributed to plasma lysophosphatidylcholine. Studies on synthetic lysophosphatides showed that polar head group structure, acyl chain length, and stereochemical configuration are important determinants for attractant activity.

Topics: Animals; Cells, Cultured; Chemotaxis, Leukocyte; Concanavalin A; Humans; Lymphocytes; Lymphoma; Lysophosphatidylcholines; Mice; Spleen; Structure-Activity Relationship

The suppressed T lymphocyte response occurring during lymphoma growth in mice was largely due to the generation of suppressor macrophages which released high amounts of prostaglandin E2 (PGE2). Based on these findings, the role of macrophage-derived PGE2 as a regulating and potentially immunosuppressive agent of the immune response is discussed.

Topics: Animals; Caseins; Cell Separation; Concanavalin A; Dinoprostone; Immunosuppressive Agents; Lymphocyte Activation; Lymphoma; Macrophages; Male; Mice; Mice, Inbred DBA; Prostaglandins E; Spleen; T-Lymphocytes

A specific antibody against myosin light chain kinase (MLCK) was used to identify the presence of a Ca2+-calmodulin-activated MLCK in mouse 1-lymphoma cells. With a double immunofluorescence technique, MLCK was determined to be accumulated directly under Con A-capped structures in a manner similar to that of previously described accumulation of actomyosin. The lymphocyte MLCK was phosphorylated in the uncapped cell and, by immunoprecipitation with a specific MLCK antibody, was shown to possess a Mr of 130,000. The MLCK was also found to constitute a major fraction of the phosphoproteins present in the plasma membrane associated-cytoskeleton. Myosin light chain kinase catalyzed the phosphorylation of both endogenous lymphocyte myosin light chains and those from smooth and skeletal muscle. The enzyme activity was dependent on the presence of Ca2+-calmodulin and was inhibited by the calmodulin-binding drug, trifluoperazine. These data suggest that the membrane-cytoskeleton-associated MLCK activity may be important in regulation of the actinmyosin contraction which is believed to be required for the collection of surface receptors into capped structures.

Topics: Animals; Calmodulin; Cell Line; Cell Membrane; Concanavalin A; Cytoskeleton; Immunologic Capping; Lymphoma; Mice; Myosin-Light-Chain Kinase; Myosins; Protein Kinases; T-Lymphocytes

Topics: Animals; Antilymphocyte Serum; Cells, Cultured; Clone Cells; Concanavalin A; Cytotoxicity Tests, Immunologic; Female; Humans; Interleukin-2; Killer Cells, Natural; Leukemia, Experimental; Lymphoma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred CBA; Mice, Nude; T-Lymphocytes, Cytotoxic; Time Factors; Trypsin

Topics: Animals; Binding, Competitive; Concanavalin A; Cytotoxicity, Immunologic; Female; In Vitro Techniques; Lymphocyte Activation; Lymphoma; Macrophage Activation; Neoplasms, Experimental; Rats; Rats, Inbred WF

Monocyte functions were studied in 16 patients with Hodgkin's disease, 11 untreated and five in unmaintained complete remission. Eleven untreated patients with non-Hodgkin's lymphomas and 21 healthy persons were used as controls. Monocytes were isolated from peripheral blood and enriched to greater than 90%. Lymphoma monocytes showed normal ability to lyse human RBC coated with anti-D IgG antibodies as evaluated by a 51Cr-release assay. The ability of monocytes to augment or suppress concanavalin A stimulation of lymphocytes purified to greater than 98% was studied by incubation of a number of lymphocytes with increasing amounts of purified monocytes. The incorporation of 14C-thymidine was potentiated by a factor of 10 in the presence of equal amounts of monocytes. There was no difference between monocytes from Hodgkin, non-Hodgkin or healthy controls to augment patients' autologous or normal lymphocytes. Patient monocytes also suppressed the response at the same monocyte-lymphocyte ratio as normal monocytes. Stimulation of patient lymphocytes without the addition of monocytes was usually lower than that of normal control lymphocytes. The difference between patient and control lymphocyte stimulation was preserved in the presence of monocytes. It is concluded that monocytes from patients with active Hodgkin's disease or non-Hodgkin's lymphoma have normal helper and suppressor effects on patient or normal lymphocytes stimulated by Con A and normal antibody-dependent cytotoxicity.

Topics: Antibody-Dependent Cell Cytotoxicity; Concanavalin A; Hemolysis; Hodgkin Disease; Humans; Lymphocyte Activation; Lymphoma; Monocytes; Phagocytosis

Topics: Adolescent; Adult; Aged; Concanavalin A; Female; Humans; Immunity, Cellular; Immunosuppression Therapy; Lymphoma; Male; Middle Aged; T-Lymphocytes; T-Lymphocytes, Regulatory

Topics: Concanavalin A; Female; Humans; Immunoglobulins; Immunosuppression Therapy; Lectins; Lymphocyte Culture Test, Mixed; Lymphocytes; Lymphoma; Middle Aged; Receptors, Antigen, B-Cell; Rosette Formation; T-Lymphocytes

The proliferative responses of peripheral blood lymphocytes of inbred SJL/J mice to the mitogens phytohemagglutinin and concanavalin A were evaluated serially in individual animals before and after induction of lymphomas by transplantation. Tumor-bearing mice exhibited marked suppression of mitogen responsiveness. Proliferative responses in mixed lymphocyte-tumor cell culture and mixed lymphocyte culture were also suppressed. Suppression of responses was tumor related, and responsiveness was restored in animals whose tumors regressed. There was no deficiency of responder T-cells in the peripheral blood of tumor-bearing animals, and suppressor cell activity was not demonstrable in a number of in vitro assays. However, plasma from tumor-bearing animals exhibited potent immunosuppressive activity. The presence of plasma suppressive activity was similarly tumor related. Thus this study demonstrates nonspecific suppression of cell-mediated immune responses following tumor induction, apparently mediated by a plasma suppressor factor.

Topics: Animals; Concanavalin A; Female; Immune Tolerance; Immunity, Cellular; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphoma; Mice; Mice, Inbred Strains; Neoplasm Transplantation; Neoplasms, Experimental; Phytohemagglutinins; T-Lymphocytes, Regulatory

Procedures are described for the isolation of a mast cell growth factor (MCGF) from medium conditioned by mitogen-activated splenic leukocytes (CM). Although optimal conditions for the production of MCGF in CM are identical to those for the production of T cell growth factor (TCGF), MCGF can be dissociated from TCGF after the first stage of purification on a DEAE-cellulose column. MCGF elutes from the column in the breakthrough fraction, whereas TCGF binds avidly to DEAE and is eluted only at high salt concentration. MCGF also differs from TCGF with respect to m.w. (as estimated by Sephadex G-150 chromatography) and sensitivity to trypsin. In addition, MCGF is produced by the murine myelomonocytic leukemia WEHI-3 and the radiation induced thymic lymphoma LBRM-33 cells, whereas TCGF is produced only by the latter in the presence of a mitogen. Another hemopoietically active factor, granulocyte colony-stimulating factor (G-CSF) present in media conditioned by WEHI-3 and LBRM-33 cells, however, shares a number of properties with MCGF. Although studies with purified or partially purified MCGF have thus far failed to reveal a correlation between MCGF and G-CSF, further biochemical analyses are necessary to dissociate MCGF from G-CSF.

Topics: Animals; Cell Differentiation; Cells, Cultured; Chromatography, DEAE-Cellulose; Chromatography, Gel; Colony-Stimulating Factors; Concanavalin A; Female; Growth Substances; Interleukin-3; Leukemia, Myeloid; Lymphoma; Male; Mast Cells; Mice; Mice, Inbred DBA; Mice, Nude; Time Factors

In this paper we report the effect of cis-dichlorodiammine platinum(II) (cis-platin) on the agglutination of normal and transformed lymphocytes with lectins conconavalin A (Con A) and wheat germ agglutinin (WGA) which is a sensitive test to study the expression of cell surface components. Normal splenocytes showed small degree of cell agglutination with Con A and WGA. This cell agglutination of normal splenocytes with these lectins increases significantly after cis-platin treatment at 37 degrees C. The Dalton's lymphoma cells which show a high degree of cell agglutination with Con A and WGA when treated with cis-platin at 37 degrees C results in decrease in the degree of their cell agglutination with these lectins. Agglutination inhibition studies with alpha-methyl-D-glucopyranoside for Con A and N-acetyl-D-glucosamine for WGA shows that this increase and decrease of cell agglutination after platinum treatment of cells is a specific reaction and not non-specific. Incubation of cis-platin treated normal splenocytes with excess of neuraminic acid results in decrease of cell agglutination with both Con A and WGA. cis-Platin treatment of the cells results in the release of sialic acid from the surface of the cells. A correlation between the release of sialic acid from the cell surface and cell agglutination is also discussed.

Topics: Animals; Cell Aggregation; Cell Membrane; Cisplatin; Concanavalin A; In Vitro Techniques; Lectins; Lymphocytes; Lymphoma; Mice; Mice, Inbred DBA; Neoplasms, Experimental; Receptors, Mitogen; Sialic Acids; Wheat Germ Agglutinins

A current hypothesis related to non-Hodgkin's lymphoma states that the wide variety of cytologic types in this disorder reflects morphologic alterations during different stages (G1, S, and G2) of the cell cycle involved in the blastogenic transformation of normal lymphocytes. In our investigations of biochemical and structural changes during lymphocyte transformation, we have used correlated stereologic morphometric analysis, assessment of chromatin organization, and autoradiography of human peripheral T-lymphocytes labeled with 3H-thymidine and stimulated with concanavalin A. These studies have confirmed that the characteristic increase in nuclear size and disaggregation of condensed chromatin masses precedes and is independent of DNA synthesis. Since the full range of morphologic alterations observed in lymphocyte transformation can occur in the G1 phase of this process, modifications to the above hypothesis are required. Assessment of the nuclear contour index following mitogen stimulation indicates that at least in this in vivo system, there is no cleaved or convoluted phase during the transformation of human peripheral T lymphocytes.

Topics: Autoradiography; Cell Division; Cell Nucleus; Concanavalin A; DNA Replication; Humans; Lymphocyte Activation; Lymphoma; Rosette Formation; T-Lymphocytes

Cultures of murine T lymphocytes with cytotoxic activity towards syngeneic RBL-5 lymphoma cells were obtained from spleen cells of immunized animals after co-culture in vivo with irradiated RBL-5 cells. At different times after initiation, these mixed tumour-lymphocyte cultures (MTLC) were multiplied by transfer to conditioned medium (CM) containing T cell growth factor (TCGF) activity, produced by the stimulation of rat spleen cells with Con A. The effect of residual Con A was investigated by the addition of specific blocking sugar, alpha-methyl mannoside (alpha MM), to the CM in some experiments. This procedure did not reduce the growth potential of the cells, and resulted in a dramatic increase in the cytotoxic activity of the cultures as measured by a 4-hr 51Cr-release assay. The cultures multiplied 1 X 10(3)-fold over a 3-week period with retention of cytotoxicity for RBL-5 cells at levels up to 70-fold greater than those of the MTLCs from which they were derived. The cultured cells, when injected i.p. together with RBL-5 cells into normal mice, were shown to mediate a significant prolongation of the survival of the treated animals. This effect was, however, less dramatic than had been expected from the in vitro results. It would therefore appear that, while cells grown in tissue culture using Con A-conditioned medium may fulfill some theoretical requirements for the immunotherapy of experimental tumours, other factor(s) are required for full protection.

Topics: Animals; Cell Division; Cells, Cultured; Concanavalin A; Culture Media; Cytotoxicity, Immunologic; Growth Substances; Lymphoma; Male; Mice; Mice, Inbred C57BL; Neoplasms, Experimental; Rats; Spleen; T-Lymphocytes

Monoclonal antibodies recognizing human T cell differentiation antigens were used to study lymphocyte populations in three cutaneous diseases. Neoplastic lymphocytes from patients with varying phases of cutaneous T cell lymphoma (mycosis fungoides, Sezary syndrome and related presentations) were reactive with OKT1 and OKT3 (pan T cell reagents) and OKT4 (an antibody defining the functional "helper" T cell subset). The malignant cells lacked membrane antigens reactive with OKT5 and OKT8 (markers of "suppressor" T cells). The presence of an OKT1+, OKT3+, OKT4+, OKT5-, OKT8- phenotype on the neoplastic T lymphocytes of cutaneous T cell lymphoma (CTCL) supports the clinical impression that all phases of CTCL represent a single disease entity. A patient with pemphigus vulgaris, a disease of autoreactive, antiepidermal antibodies was shown to consistently have a marked expansion of the peripheral blood OKT4 reactive T lymphocyte population. These findings suggest that autoantibodies in pemphigus vulgaris may occur in the context of a profound OKT4/OKT5 immunoregulatory imbalance. Peripheral blood lymphocytes from patients ith extensive psoriasis vulgaris had a normal profile of reactivity with the OKT antibodies. In addition, OKT6 (marker of intrathymic T cells) has been shown to react with Ia+ dendritic cells in the epidermis suggesting that this antibody may recognize Langerhans' cells.

Topics: Adolescent; Adult; Aged; Animals; Antibodies, Monoclonal; Cell Transformation, Neoplastic; Concanavalin A; Humans; Lymph Nodes; Lymphocytes; Lymphoma; Mice; Middle Aged; Pemphigus; Phenotype; Psoriasis; Rabbits; Skin Diseases; T-Lymphocytes

Topics: Candida albicans; Candidiasis, Oral; Central Nervous System Diseases; Child, Preschool; Concanavalin A; Eczema; Epithelium; Humans; Infant; Lymphoma; Male; Phytohemagglutinins; Pokeweed Mitogens; Recurrence; Rosette Formation; Thymus Gland; Wiskott-Aldrich Syndrome

Topics: Animals; Antigens, Surface; Cell Line; Cell Survival; Concanavalin A; Edetic Acid; Lymphoma; Mast-Cell Sarcoma; Mice; Mice, Inbred Strains; Spleen; T-Lymphocytes; Time Factors

Serum IgE levels were evaluated in 119 untreated and 112 treated patients with Hodgkin's disease (HD). 38 of the nonatopic untreated patients showed significantly increased (> 300 IU/ml) IgE concentrations. No relationship could be found between increased IgE levels and depressed lymphocyte response to phytohemagglutinin (PHA) or the imbalance of TM and TG lymphocyte subsets. On the other hand, the mean level of suppressor activity elicitable from cells of untreated HD patients by concanavalin A preincubation did not differ significantly from that of healthy control subjects. In contrast, in treated patients, where there was a significant reduction in the number of circulating T lymphocytes, a further depression of the lymphocyte response to PHA, a more marked disproportion of TM and TG cell subsets and a noticeable fall in IgE concentration was found. These data suggest that increased IgE concentrations seen in untreated patients with HD are unrelated to the T-cell defects. They also suggest that hyperproduction of IgE is probably not invariably a consequence of a suppressor cell deficiency.

Topics: Cell Division; Concanavalin A; Hodgkin Disease; Humans; Immunoglobulin E; Lymphocytes; Lymphoma; T-Lymphocytes

Of 12 T-lymphoma cell lines investigated, one line, EL-4 azgr (Thy-1+, Lyt-1+, Lyt-2-3-,Ia-, T-200+, sIg-, and FcR-) and, to a lesser extent, the parental cell line, EL-4, produced T cell growth factor(s) (TCGF) when stimulated by the T-cell mitogen concanavalin A (Con A). Induced production of TCGF-E was detected by 6 h and maximal at 18-24 h. Purified TCGF-E from this source had an approximately 30,000 mol wt and the biological activity of TCGF produced by whole spleen cells, including: augmentation of T cell-mitogen responses, cytotoxic T lymphocyte (CTL) proliferation support dependence, augmented generation of CTL, lack of strain specificity, and failure to stimulate resting T cells. TCGF-E is neither synthesized or secreted by this lymphoma cell line unless stimulated by Con A. X-irradiation up to 7,000 rad failed to inhibit synthesis and secretion. These observations have a practical application in providing a relatively homogeneous clonal cell product for T cell culture support and for structural and functional studies of the TCGF molecule(s). They suggest also a model for examining mechanisms of triggering production and secretion of a regulatory molecule that controls T cell functions.

Topics: Animals; Cell Line; Concanavalin A; Growth Substances; Lymphocyte Activation; Lymphoma; Mice; Molecular Weight; Spleen; T-Lymphocytes

Topics: Animals; Cell Division; Concanavalin A; Cytotoxicity, Immunologic; Epitopes; In Vitro Techniques; Lymphoma; Male; Mast-Cell Sarcoma; Mice; Mice, Inbred Strains; Neoplasm Transplantation; Neoplasms, Experimental; Protein Biosynthesis; T-Lymphocytes; Transplantation, Homologous

Antisera against the C57B1 (H-2b) mouse lymphoma, EL4 were prepared in rabbits. After absorption with mouse liver, red cells and thymocytes the antisera appeared to be cytotoxic for a subpopulation of peripheral T cells. The absorbed antisera blocked the immunosuppressor function of Con A-stimulated splenic lymphocytes, but was unreactive against Con A-stimulated and allogeneically primed cytotoxic cells, or helper T cells. Consequently, heteroantiserum against EL4 may provide a useful reagent for the differentiation of cytotoxic from suppressor T-cell subsets.

Topics: Animals; Antibody Formation; Cells, Cultured; Concanavalin A; Cytotoxicity, Immunologic; Fluorescent Antibody Technique; Immune Sera; Lymphoma; Mice; Mice, Inbred C57BL; Neoplasms, Experimental; Rabbits; Spleen; T-Lymphocytes, Regulatory

Topics: Animals; Chlorides; Concanavalin A; Cytotoxicity, Immunologic; Humans; Hydrogen Peroxide; Iodides; Iodine Radioisotopes; Lymphoma; Methylmannosides; Mice; Mice, Inbred BALB C; Neutrophils; Peroxidase; Peroxidases

Topics: Animals; Antibodies; Concanavalin A; Cytotoxicity, Immunologic; Female; H-2 Antigens; Immune Sera; Immunologic Capping; Lymphoma; Male; Mast-Cell Sarcoma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred DBA; Parainfluenza Virus 1, Human; T-Lymphocytes

The HPRS line 1 lymphoblastoid cells derived from an ovarian lymphoma induced by Marek's disease virus in chickens displayed increased agglutinability and haemadsorption when incubated with Con A. The increase was proportional to the concentration of Con A used. The same Con A-untreated cells and normal chicken lymphocytes incubated with Con A exhibited no increase in agglutinability and haemadsorption, however.

Topics: Agglutination; Animals; Cell Line; Cell Membrane; Chickens; Concanavalin A; Erythrocytes; Female; Hemadsorption; Herpesvirus 2, Gallid; Humans; Lymphoma; Ovarian Neoplasms; Receptors, Concanavalin A

Topics: Animals; Concanavalin A; Cytotoxicity, Immunologic; Lymphoma; Mice; Neoplasms, Experimental; T-Lymphocytes

Topics: Concanavalin A; Humans; Immunity, Cellular; Lymphocyte Activation; Lymphoma; Phytohemagglutinins

Topics: Animals; Concanavalin A; Electrophoresis; Lymphocytes; Lymphoma; Mice; Neoplasms, Experimental

A patient with Sézary syndrome developed a diffuse undifferentiated lymphoma of T-cell origin. After becoming resistant to multiple chemotherapeutic agents, the patient was treated with antithymocyte globulin. A 75% reduction in adenopathy and complete resolution of skin erythema was observed during an 8-day period. In addition the percent of circulating T cells and the ability of those cells to respond to phytohemagglutinin and concanavalin A were reduced after antithymocyte globulin therapy. The patient died of an intracerebral hemorrhage secondary to profound thrombocytopenia. The study suggests that tumor lysis may be achieved by passive antibody therapy in certain advanced lymphomas.

Topics: Antilymphocyte Serum; Concanavalin A; Dermatitis, Exfoliative; Humans; Immunotherapy; Keratoderma, Palmoplantar; Lectins; Lymphatic Diseases; Lymphocyte Activation; Lymphoma; Male; Middle Aged; Syndrome; T-Lymphocytes

Previously, 10 BALB/c T cell lines (Thy 1.2+, Ig-) were shown to express different combinations of Ly 1 and Ly 2 antigens. The possible immunologic function(s) of these tumor cells was determined by investigating the effects of these cells on the responses to mitogens, the mixed lymphocyte response (MLR), and the generation of cell-mediated lysis (CML) by normal spleen cells. Five T cell lines, P1798 and BALENTL 3, 5, 8, and 9, continued to synthesize DNA after exposure to large doses of irradiation. Only BALENTL 4, 6, 7, and 14 (Ly 1-(2+)) and BALENTL 13 (Ly 1+(2-)) were radiosensitive and therefore amenable to study. BALENTL 4 and 14 gave significant suppression of the MLR between BALB/c and C57BL/6; BALENTL 14 also inhibited the generation of BALB/c effector cells against C57BL/6 spleen cells. None of these T cell lines had any effect on the proliferative response of BALB/c spleen cells induced by concanavalin A. However, there was approximately 50% suppression of the phytohemagglutinin response of BALB/c spleen cells by BALENTL 14.

Topics: Animals; Cell Line; Concanavalin A; Cytotoxicity, Immunologic; Female; Immunity, Cellular; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphoma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Phytohemagglutinins; T-Lymphocytes; Thymidine

Topics: B-Lymphocytes; Cell Aggregation; Cell Line; Concanavalin A; Glycoproteins; Humans; Lymphoma; Membrane Proteins; Sialic Acids; Sialyltransferases; T-Lymphocytes; Transferases

Topics: Animals; Cell Cycle; Concanavalin A; Fluorescent Antibody Technique; Immunologic Capping; Lymphocyte Activation; Lymphocytes; Lymphoma; Membrane Fluidity; Mice; Microtubules; Receptors, Antigen, B-Cell; Tubulin

The dynamics of the toxin Ricinus communis agglutinin II (RCAII or ricin) on cells of a murine lymphoma line (BW5147) and a toxin-resistant variant line (BW5147RicR.3) that is 200 times more resistant than the parent to direct RCAII cytotoxicity were examined using ferritin-conjugated, affinity purified, 125I-labeled RCAII (ferritin-125I-RCAII). Ferritin-125I-RCAII was indistinguishable from native RCAII in quantitative binding and cytotoxicity experiments. When RCAII-sensitive BW5147 and -resistant BW5147RicR.3 cells were labeled with ferritin-125I-RCAII at various toxin concentrations (1--10 microgram/ml), no differences in toxin binding were observed. These same cells were examined by electron microscopy. At low ferritin-125I-RCAII concentrations (1-3 microgram/ml RCAII) where only the parental BW5147 cells were significantly more sensitive to RCAII, toxin receptors were internalized by ferritin-125I-RCAII-induced endocytosis. In parallel experiments, ferritin-125I-RCAII that bound to the resistant BW5147RicR.3 cells remained relatively dispersed or clustered, and there was little evidence of transport into cells via endocytosis. At higher ferritin-125I-RCAII concentrations (greater than 7 microgram/ml RCAII) where both parental and resistant variant cells are sensitive to the cytotoxic effects of RCAII, more ferritin-conjugated toxin was bound, and subsequent endocytosis occurred to a similar degree in both cell types. Endocytosis of ferritin-conjugated concanavalin A was indistinguishable on RCAII-sensitive parental and resistant variant cells at all concentrations tested. The results suggest that a specific defect on the selected BW5147RicR.3 cells prevents RCAII entry into these cells a low toxin concentrations, rendering them more resistant to the cytotoxic effects of RCAII.

Topics: Animals; Cell Line; Concanavalin A; Drug Resistance; Endocytosis; Ferritins; Genetic Variation; Iodine Radioisotopes; Lymphoma; Mice; Neoplasms, Experimental; Receptors, Concanavalin A; Receptors, Drug; Ricin

Topics: Adsorption; Agglutination; Animals; Cells, Cultured; Concanavalin A; Dacarbazine; Female; Leukemia L1210; Lymphoma; Male; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Triazenes

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

Peripheral blood lymphocytes and their in vitro reactivity have been recorded prior to treatment in 18 patients with Hodgkins disease, 11 with lymposarcoma, 13 with reticulosarcoma, 20 with various solid tumors and 37 normal control persons. The mean total numbers of lymphocytes, those of T lymphocytes,and the mean reactivity to PHA and ConA were reduced in all groups except lymphosarcoma, although with varying degrees of statistical significance. The percentages of T and B lymphocytes appeared to be normal in all groups, but the ranges of values were somewhat greater than among the normal controls. The mean total numbers of B lymphocytes were normal in all patient groups. All reductions seemed to be more pronounced in patients with disseminated than in those with localized disease, but none of these differences was statistically significant. All patient groups appeared to have reduced reactivity in MLC, while the ability to stimulate control lymphocytes was nearly normal. The results fail to indicate an in vitro immunological abberation specific to Hodgkin's disease. It seems that human malignant, neoplastic diseases are associated with a relatively selective reduction of the total numbers and reactivity of blood T lymphocytes. Various explanations of the reactivity impairment are proposed. The pathogenesis of the reduction of the total number of blood T lymphocytes remains obscure.

Topics: Adult; Aged; B-Lymphocytes; Concanavalin A; Female; Hodgkin Disease; Humans; Immune Adherence Reaction; Immunity, Cellular; Immunoglobulins; Lectins; Leukocyte Count; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphoma; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Male; Middle Aged; Mitogens; T-Lymphocytes

Topics: Adenocarcinoma; Animals; Cell Survival; Cell-Free System; Concanavalin A; Cytopathogenic Effect, Viral; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocytes; Lymphoma; Mammary Neoplasms, Experimental; Rats; Rats, Inbred BN; Rats, Inbred F344; Rats, Inbred WF; Spleen; Thymidine; Ultracentrifugation; Ultrasonics; Ultraviolet Rays

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 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

Injections of appropriate numbers of irradiated tumor cells produced antibodies against tumor cell-surface antigen(s) in both syngeneic tumor models studied: the early transplant generations of the spontaneous L2 lymphoma in AKR/J mice and the chemically induced EL 4 lymphoma in C57BL/6J mice. No antibody was detected in normal or nonimmunized tumor-bearing mice. Tumor inhibitory or enhancing activity was not demonstrated by these antibodies. Immunoprophylaxis or cell-mediated immunity against the L2 lymphoma was not observed after injections of irradiated L2 cells and/or BCG into AKR mice. However, injections of irradiated EL 4 cells alone were effective in immunoprophylaxis against as many as 10(6) EL 4 cells and in immunotherapy against 10(2) EL 4 cells per mouse. The addition of BCG injections made immunotherapy with irradiated EL 4 cells effective against a load of 10(4) EL 4 cells/mouse, though BCG alone was not effective for immunoprophylaxis against EL 4 cells. Resistance to EL 4 could be transferred with viable syngeneic peritoneal or nucleated spleen cells. In both tumor models, an ongoing delayed hypersensitivity reaction to BCG alone apparently did not inhibit bystander tumor cells even when tumor cells were mixed before inoculation with viable BCG. In neither tumor model were concanavalin A-coated tumor cells more potent for immunoprophylaxis than were irradiated tumor cells alone.

Topics: Animals; Antibodies, Neoplasm; Antibody Formation; Antigens, Neoplasm; BCG Vaccine; Concanavalin A; Female; Granuloma; Immunity, Cellular; Immunization; Immunotherapy; Lymphocytes; Lymphoma; Mice; Mice, Inbred AKR; Mice, Inbred C57BL; Mycobacterium bovis; Neoplasms, Experimental

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

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

Lymphocytes 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

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

The ability of cells to bind to nylon fibers coated with lectin molecules interspaced with varying numbers of albumin molecules has been analyzed. The cells used were lymphoma cells, normal lymphocytes, myeloid leukemia cells, and normal and transformed fibroblasts, and the fibers were coated with different densities of concanavalin A or the lectins from soybean or wheat germ. Cells fixed with glutaraldehyde did not bind to lectin-coated fibers. The number of cells bound to fibers could be increased by increasing the density of lectin molecules on the fiber, the density of specific receptors on the cell, or the mobility of the receptors. It is suggested that binding of cells to fibers involves alignment and binding of specific cell surface receptors with lectin molecules immobilized on the fibers, and that this alignment requires short-range rapid lateral mobility (RLM) of the receptors. The titration of cell binding to fibers coated with different densities of lectin and albumin has been used to measure the relative RLM of unoccupied cell surface receptors for the lectin. The results indicate a relationship of RLM to lectin-induced cell-to-cell binding. The RLM or receptors for concanavalin A (Con A) was generally found to be higher than that of receptors for the lectins from wheat germ or soybean. Receptor RLM could be decreased by use of metabolic inhibitors or by lowering the temperature. Receptors for Con A had a lower RLM on normal fibroblasts than on SV40-transformed fibroblasts, and trypsinization of normal fibroblasts increased Con A receptor RLM. Normal lymphocytes, lymphoma cells, and lines of myeloid leukemia cells that can be induced to differentiate had a high receptor RLM, whereas lines of myeloid leukemia cells that could not be induced to differentiate had a low receptor RLM. These results suggest that the RLM of Con A receptors is related to the transformation of fibroblasts and the ability of myeloid leukemia cells to undergo differentiation

Topics: Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Dinitrophenols; Fibroblasts; Glycine max; Lectins; Leukemia, Myeloid; Lymphocytes; Lymphoma; Mice; Models, Biological; Nylons; Plant Lectins; Rats; Serum Albumin, Bovine; Simian virus 40; Temperature; Triticum

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: Antigens, Neoplasm; Antigens, Viral; Binding Sites, Antibody; Cell Line; Concanavalin A; Herpesvirus 4, Human; Hodgkin Disease; Humans; Lymphatic Diseases; Lymphoma

The cell-to-cell binding induced by concanavalin A (Con A) and the lectins from wheatgerm, soybean, and waxbean has been analyzed by measuring the ability of single cells to bind to lectin-coated cells immobilized on nylon fibers. The cells used were lymphoma, myeloid leukemia, and normal fibroblast cells. With all lectins, cell-to-cell binding was inhibited if both cells were prefixed with glutaraldehyde. However, in most cases cell-to-cell binding was enhanced when only the lectin-coated cell was prefixed. With normal fibroblasts, treatment of either one or both cells with trypsin enhanced the cell-to-cell binding induced by Con A and the wheatgerm lectin. Neuraminidase, which increases the number of receptors for soybean agglutinin, increased cell-to-cell binding only if both cells were treated. Although cell-to-cell binding induced by the lectins from soybean and wheatgerm could be partially reversed by the appropriate competitive saccharide inhibitor, binding induced by Con A could not be reversed. The experiments indicate that cell-to-cell binding induced by a lectin can be prevented by an insufficient density of receptors for the lectin, insufficient receptor mobility, or induced clustering of receptors. These effects can explain the differences in cell-to-cell binding and agglutination observed with different cell types and lectins. They also suggest that cell-to-cell binding induced by different lectins with a variety of cell types is initiated by a mechanism involving the alignment of complementary receptors on the colliding cells for the formation of multiple cell-to-lectin-to-cell bridges.

Topics: Agglutination; Animals; Binding Sites, Antibody; Cell Adhesion; Cell Line; Cells, Cultured; Clone Cells; Concanavalin A; Cricetinae; Fibroblasts; Glutaral; Glycine max; Lectins; Leukemia, Myeloid; Lymphoma; Mice; Models, Biological; Neuraminidase; Nylons; Plant Lectins; Triticum; Trypsin; Vegetables

Concanavalin A (Con A) induces movement of its receptors on the cell surface membrane. This induction results in a concentration of Con A site complexes on one pole of the cell to form a cap. A marked difference was found in the mobility of Con A receptor between lymphocytes from normal persons and lymphocytes from patients with Hodgkin's disease and other malignant lymphomas. Lymphocytes isolated from tonsils of patients undergoing tonsillectomy and from axillary lymph nodes of breast cancer patients exhibited approximately 30% of cells with caps, which is identical with the cap formation ability of normal lymphocytes. In biopsy material from patients with Hodgkin's disease and other malignant lymphomas, a significant decrease in the ability of the lymphocytes to form caps was observed. This difference in the mobility of Con A sites was even more pronounced in lymphocytes isolated from the peripheral blood. In 123 patients with Hodgkin's disease and other malignant lymphomas, cap formation ranged between 3 and 12%. The ability of cells, from a normal donor or a lymphoma patient, to form caps was independent of the source from which the lymphocytes were isolated, e.g., lymph node, spleen, or blood. Lymphocytes from patients with lymphoma were also agglutinated by Con A to a higher degree than normal lymphocytes. These findings are discussed in relation to the association of the lymphocytes with these malignancies and as a possible aid in their differential diagnosis.

Topics: Agglutination; Binding Sites, Antibody; Cell Membrane; Concanavalin A; Hodgkin Disease; Humans; Lymph Nodes; Lymphocytes; Lymphoma; Lymphoma, Non-Hodgkin; Palatine Tonsil

Maintenance of remission solely by repeated BCG vaccinations in seven patients with non-Hodgkin's lymphoma who had achieved a complete clinical remission with initial standard therapy has provided sufficient encouragement to begin a randomized clinical trial. In vitro lymphocyte responses to mitogens and PPD used as parameters of cell-mediated immunity have not proved to be of value in predicting early or late recurrence in six pre-trial and trial patients. Eight out of twenty-one patients with malignant melanoma have shown a satisfactory clinical response (10-34 months) to immunotherapy. Those who respond must show immunological reactivity to the stimulating agent, however the best clinical responses were not associated with the highest degrees of in vivo and in vitro sensitization. The skin reactivity and the in vitro lymphocyte response to PPD as well as a 2-3-fold increase in the appearance of colony-forming units in the peripheral blood following the intratumour injection of BCG or PPD are helpful in prognosis and management of these patients. All patients with malignant melanoma who presented with a PHA response less than 40% of normal made a poor response to immunotherapy. Autopsies performed on seven patients dying with extensive melanocarcinomatous disease failed to show any serious adverse toxic reactions or infections from oral and intratumour injections of BCG.

Topics: Administration, Oral; Adult; BCG Vaccine; Concanavalin A; DNA; Female; Hematopoietic Stem Cells; Humans; Immunoglobulins; Immunotherapy; Injections, Intradermal; Lectins; Lymphocyte Activation; Lymphoma; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Melanoma; Skin Neoplasms; Tuberculin; Vaccinia virus

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

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

Topics: Adenosine Triphosphate; Animals; Cell Aggregation; Cell Division; Cell Line; Concanavalin A; Deoxy Sugars; Deoxyglucose; Fucose; Glucosamine; Glycoproteins; Lectins; Lymphoma; Mice; Sialic Acids; Time Factors

Sera from mice with transplanted 3-methylcholanthrene (MCA)-induced tumors inhibited the proliferative response of normal mouse lymphocytes to mitogens and allogeneic cells. The sera were not toxic to these cells and did not inhibit mitogen responses by an increased binding of mitogen to serum components. The sera could have prevented the initiation of the proliferative response or could have inhibited cells already proliferating. The uptake of 3H-uridine, an event preceding DNA synthesis, was also suppressed. The sera had no effect on proliferation of a normal tissue culture line or an allogeneic tumor cell line induced by MCA. However, sera from tumor-bearing mice slowed growth of the autochthonous tumor cells and allogeneic lymphoma cells but did not completely block their proliferation. Exposure of lymphoid cells to the sera for a period as brief as 1 hour markedly decreased the ability of cells to respond subsequently to mitogens, and washing of the cells did not restore that ability. The inhibitory activity of the sera was only partially removed by absorption with lymphoid cells or cell lines.

Topics: Animals; Cell Line; Concanavalin A; Lipopolysaccharides; Lymphocyte Activation; Lymphocytes; Lymphoma; Mice; Mice, Inbred Strains; Neoplasms, Experimental; Rhabdomyosarcoma; Thymidine; Uridine

Topics: Animals; Cell Line; Cell Membrane; Cells, Cultured; Complement System Proteins; Concanavalin A; Cytotoxicity Tests, Immunologic; Disease Models, Animal; Erythrocytes; Fibroblasts; Fluorescent Antibody Technique; Haplorhini; Herpesviridae; Immune Adherence Reaction; In Vitro Techniques; Interferons; Lectins; Lymphocyte Activation; Lymphocytes; Lymphoma; Lymphotoxin-alpha; Mitogens; Monkey Diseases; Oncogenic Viruses

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; Animals; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Culture Techniques; Electricity; Ferritins; Fibroblasts; Lymphocytes; Lymphoma; Lysine; Models, Biological; Moloney murine leukemia virus; Neoplasms, Experimental; Neuraminidase; Polymers; Rats; Simian virus 40; Surface Properties

Topics: Animals; Cell Line; Cell Membrane; Cell Nucleus; Concanavalin A; Freeze Etching; Lymph Nodes; Lymphocytes; Lymphoma; Membranes; Mice; Mice, Inbred C57BL; Microscopy, Electron; Swine; Temperature; Thymus Gland

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

The cell-cell binding induced by concanavalin A between single cells has been analyzed by use of cells attached to nylon fibers. Binding of a concanavalin A-coated cell to an untreated cell was found to a high degree between two lymphoma tumor cells, less frequently between a lymphoma cell and a normal lymphocyte, and only rarely between two normal lymphocytes. The binding was inhibited by the presence of a saccharide inhibitor of concanavalin A, but could not be reversed by addition of the inhibitor after the cells had bound to each other. Although no binding was obtained when both cells were coated with lectin or fixed with glutaraldehyde, fixation of a cell before coating with concanavalin A enhanced its ability to bind an untreated cell. The results indicate that cell-cell binding induced by concanavalin A requires short-range lateral movement of cell receptors for the lectin, that only one cell has to have mobile receptors, and that some receptors must be unoccupied by lectin molecules before cell-cell contact. Clustering of the receptors is not necessary and seems to hinder cell-cell binding. It is suggested that the short-range movement is required for alignment of individual receptors so as to form multi-point bridges between two cells by lectin molecules. The bridging is then followed by the formation of irreversible bonds between the cells. The receptors on tumor cells appear to have a greater ability than receptors on normal cells to align themselves for cell-cell binding.

Topics: Agglutination; Animals; Antibodies, Neoplasm; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Leukemia, Experimental; Lymphocyte Activation; Lymphocytes; Lymphoma; Mice; Mice, Inbred A; Microscopy, Fluorescence; Moloney murine leukemia virus; Rats; T-Lymphocytes

Topics: Agglutination; Agglutination Tests; Animals; Binding Sites, Antibody; Carbohydrates; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fibroblasts; Fluorescent Antibody Technique; Glycogen; Leukemia, Experimental; Leukemia, Myeloid, Acute; Lymphocytes; Lymphoma; Male; Mice; Movement; Rats; Tritium; Trypsin

Topics: Absorption; AKR murine leukemia virus; Animals; Antibody-Producing Cells; Antigens; Antilymphocyte Serum; Binding Sites; Chromium Radioisotopes; Complement System Proteins; Concanavalin A; Cytotoxicity Tests, Immunologic; Erythrocytes; Hemolytic Plaque Technique; Immunity, Cellular; Immunoglobulins; Lectins; Lymphocyte Activation; Lymphoma; Perissodactyla; Rats; Rats, Inbred F344; Rats, Inbred Lew; Sheep; Spleen; T-Lymphocytes

Topics: Agglutination; Animals; Ascitic Fluid; Binding Sites; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Formaldehyde; Glutaral; Glycine max; Lectins; Lymphoma; Mice; Moloney murine leukemia virus; Plant Lectins; Polysaccharides; Protein Binding; Time Factors; Triticum; Tritium

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Diffusion; Energy Transfer; Fibroblasts; Fluorescence; Glutaral; Lymphocytes; Lymphoma; Protein Binding; Rats; Rotation; Simian virus 40; Sucrose; Trypsin

Topics: Animals; Cell Line; Concanavalin A; DNA, Neoplasm; Galactosamine; Galactose; Lectins; Leukemia, Myeloid; Lipopolysaccharides; Lymphoma; Mannose; Methylglycosides; Mice; Mitogens; Multiple Myeloma; Neoplasms, Experimental; Polysaccharides, Bacterial; Ricin; RNA, Neoplasm; Salmonella typhi; T-Lymphocytes

Topics: Animals; Binding Sites; Cell Aggregation; Cell Line; Cell Membrane; Concanavalin A; Fluoresceins; Lectins; Lymph Nodes; Lymphocytes; Lymphoma; Mice; Microscopy, Ultraviolet; Moloney murine leukemia virus; Neoplasms, Experimental; Plant Lectins; Polysaccharides; Rats; Spleen; T-Lymphocytes; Triticum; Tritium

Topics: Cell Adhesion; Cell Line; Cells, Cultured; Concanavalin A; Cycloheximide; Dactinomycin; Edetic Acid; Galactose; Glass; Glycosides; Humans; Lectins; Leukemia, Myeloid; Lymphoma; Mannose; Mast-Cell Sarcoma; Microscopy, Phase-Contrast; Plant Lectins; Temperature; Trypsin; Vegetables

Topics: Agglutination Tests; Animals; Binding Sites; Binding Sites, Antibody; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Cricetinae; Fibroblasts; Goats; Lectins; Lymphocytes; Lymphoma; Mice; Models, Biological; Neoplasms, Experimental; Protein Binding; Rats

Topics: Animals; Binding Sites; Cell Aggregation; Cell Membrane; Cell Transformation, Neoplastic; Concanavalin A; Lectins; Lymphoma; Mice; Neoplasms, Experimental; Protein Binding

Topics: Animals; Concanavalin A; DNA; Lectins; Leukemia L1210; Lymphoma; Mice; Neoplasm Transplantation; Neoplasms, Experimental; Thymidine; Thymus Neoplasms

The carbohydrate-binding protein, Concanavalin A (Con A), binds to glucose- or mannose-like sites on the cell-surface membrane. Unless the cells are treated with trypsin, this protein agglutinates malignantly transformed cells, but not normal cells. The transformed cells were agglutinated at 24 degrees C but not at 4 degrees C. Transformed and normal cells treated with trypsin were agglutinated at both 24 degrees C and 4 degrees C with high concentrations of Con A (500 mug/ml), but only at 24 degrees C with low concentrations (5 mug/ml). The same number of Con A molecules were bound to normal and transformed cells at both temperatures. The results indicate that the site for Con A on the surface membrane contains two activities, a component that binds Con A molecules (B) and a component that determines agglutination (A). B is not temperature sensitive and is active in normal and transformed cells, whereas A, which is temperature sensitive, is in an active form only in transformed cells. A can be activated by trypsin, and the increased activity per cell allows agglutination at 4 degrees C with a high, but not with a low, concentration of Con A. Agglutination of transformed cells by wheat-germ agglutinin, which binds to N-acetyl-D-glucosamine-like sites, and by soybean agglutinin, which binds to N-acetyl-D-galactosamine-like sites, was not temperature sensitive. Thus, the temperature-sensitive component A is specific for Con A, and malignant transformation of normal cells, which results in agglutinability by Con A, is associated with the activation of a specific temperature-sensitive activity on the surface membrane.

Topics: Agglutination Tests; Animals; Avian Sarcoma Viruses; Binding Sites; Cattle; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Concanavalin A; Cricetinae; Haplorhini; Hexoses; Lectins; Lymphoma; Moloney murine leukemia virus; Neoplasms, Experimental; Nitrosamines; Polyomavirus; Protein Binding; Temperature

Topics: Carcinoma; Cell Migration Inhibition; Cell Movement; Concanavalin A; Culture Techniques; Lectins; Lymphoma; Neoplasms, Experimental