concanavalin-a and Hemolysis

Topics: Animals; Animals, Newborn; Antibody Formation; Antibody-Producing Cells; B-Lymphocytes; Blood; Cell Adhesion; Cell Division; Concanavalin A; Cyclophosphamide; Erythrocytes; Hemolysis; Horses; Immune Tolerance; Mice; Radiation Chimera; Sheep; Spleen; Splenectomy; T-Lymphocytes

As of late, numerous reports have demonstrated the multiple biological activities of polyphenolic flavonoids. Amongst these reports, some indicate that the flavonoids play an important role in inflammation therapy. In this present study, we investigated the effect of rutin, a polyphenolic flavonoid, on septic arthritis due to Candida albicans, a major etiological agent that causes fungal arthritis. To induce septic arthritis, an emulsified mixture of C. albicans cell wall and Complete Freund's Adjuvant (CACW/CFA) was injected into BALB/c mice via hind footpad route once a day, everyday, for three days. In order to determine the effect of rutin, twenty-four hours after the final injection, mice having the swollen footpad were given the flavonoid (1 mg/dose/mouse) intraperitoneally every other day for three times. The footpad-edema was measured for a period of 17 days. Results showed that the rutin treatment reduced app. 45% of the edema at the peak day (day 11) of septic arthritis (P<0.05). In addition, 6 days after the peak, there was an app. 35% additional reduction of the edema (P<0.05). We found that this anti-arthritic activity was mediated by rutin's ability to inhibit nitric oxide production from macrophages and T-cells proliferation. Furthermore, this flavonoid also inhibited the growth of C. albicans yeast cells (P<0.01) and resulted in no hemolysis. These data indicate that rutin, which has both anti-arthritic and antifungal effects, can safely be administered into the blood circulation for treatment of septic arthritis caused by C. albicans. Ultimately, it can be suggested that the dual effects of rutin, anti-arthritic and anti-candidal may be helpful as an all-in-one treatment for septic arthritis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antifungal Agents; Arthritis, Experimental; Arthritis, Infectious; Candida albicans; Candidiasis; Cell Proliferation; Concanavalin A; Female; Hemolysis; Lipopolysaccharides; Macrophages; Mice; Mice, Inbred BALB C; Mitogens; Nitric Oxide; Rutin; T-Lymphocytes

Saliva and salivary gland extract (SGE) of Ixodes ricinus ticks have suppressive effects on the innate immune response of BALB/c mice. Tick saliva prevents hemolysis of sheep red blood cells (SRBC) by the human alternative pathway of complement. The adaptive immune response is also modulated by tick antigens (saliva or SGE). When stimulated in vitro with increasing doses of tick antigens, the proliferation and IL-4 production of draining lymph node T cells of mice infested with nymphal ticks increase, peak and then decrease. These results indicate that immunostimulative and immunosuppressive molecules have competing effects in tick saliva or in SGE. I. ricinus saliva inhibits, in a dose-dependent manner, splenic T cell proliferation in response to concanavalin A (ConA). Tick SGE or saliva injected intraperitoneally to BALB/c mice simultaneously with SRBC systemically immunosuppress the anti-SRBC response as shown in vitro by the reduced responsiveness of sensitized splenic T cells to restimulation with SRBC. In brief some components of SGE or tick saliva reduce the responsiveness of draining lymph node T cells and of sensitized splenic T cells in vitro. The responsiveness of naive splenic T cells to ConA stimulation in vitro is also decreased by tick saliva. Modulation of host responses by tick antigens may facilitate tick feeding, transmission and the propagation of pathogens.

Topics: Animals; Concanavalin A; Female; Hemolysis; Immunosuppression Therapy; Interleukin-4; Ixodes; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Saliva; Salivary Glands; Sheep; Spleen; Tissue Extracts

Topics: Animals; Antioxidants; Blood; Cats; Cell Division; Concanavalin A; Dietary Fats, Unsaturated; Erythrocytes; Fatty Acids, Unsaturated; Female; Ferric Compounds; Fish Oils; Hemolysis; Hydrogen Peroxide; Lymphocytes; Male; Nutritional Requirements; Oxidation-Reduction; Random Allocation; Vitamin E

The complement system is an essential part of the innate defense, and C3 is an integral part of this powerful system. In previously identified complement C3 deficient guinea pigs only approx. 5% of the normal serum C3 level is detectable. No differences were found between in vitro C3 protein synthesis and C3 mRNA levels of cells from C3-deficient and wild-type animals and the amino acid sequences of both C3 proteins are identical as deduced from cDNA sequencing. Previously, the principal inability to form a C3 thiolester was discussed as a possible reason for this C3-deficiency. Here we report the isolation of two functionally different C3 species from the C3-deficient animals. Only one of these C3 proteins exhibits normal hemolytic activity and contains a thiolester group. The second C3 species is exclusively present in C3-deficient animals and lacks a thiolester, explaining its failure to express hemolytic activity. The presence of a second C3 species lacking a thiolester structure only in C3-deficient animals indicates that the stability of the thiolester may play a role in C3 deficiency. However further analysis of the in vitro stability of the thiolesters of C3 from normal and C3-deficient guinea pigs revealed no differences. A decreased in vivo thiolester stability might lead to the presence of C3 with and without a thiolester or alternatively the expression of two isoforms of C3 in these animals. Considering the central role of C3 in host defense, the mechanisms of C3 thiolester formation require further analysis.

Topics: Animals; Biotin; Blotting, Western; Carbohydrates; Carbon Radioisotopes; Chemical Fractionation; Complement C3; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Guinea Pigs; Hemolysis; Isotope Labeling; Methylamines; Mice; Mice, Knockout; Protein Isoforms; Sodium Dodecyl Sulfate; Sulfhydryl Compounds

Due to their multivalent binding character, lectins when added exogenously will cross-link membrane surface receptors leading to lateral molecular reorganizations in the plane of the bilayer. This study reports for the first time that agglutination of rabbit erythrocytes by lentil lectin and concanavalin A increases their osmofragility. Increase in osmofragility was detected by measuring the hemolysis of erythrocytes in hypotonic as well as in isotonic solutions. It was also found that agglutination per se does not increase osmofragility but the binding of legume lectin is essential since human Rh+ cells agglutinated by a monoclonal antibody do not exhibit hemolysis.

Topics: Animals; Concanavalin A; Erythrocyte Membrane; Hemagglutination; Hemolysis; Lectins; Male; Osmotic Fragility; Rabbits

Selection of poultry for fast growth rate is often accompanied by a reduction in specific immune responses or increased disease susceptibility. In this study, 17-wk-old male turkeys from each of four closed genetic lines, a randombred control (RBC) line and its subline (F) selected for increased 16-wk BW, and another RBC and its subline (E) selected for increased egg production, were tested for in vivo response to toe web inoculation with phytohemagglutinin-P (PHA-P), in vitro response of lymphocytes in whole blood to PHA-P and concanavalin A (Con A), hemolytic complement activity, differential white blood cell counts, hematology, and serum chemistry values. Fifteen male turkeys from each of two commercial lines, Com A and Com B, were also tested. The large-bodied F line birds had a lower toe web response to PHA-P, lower lymphocyte counts, and lower relative spleen weights than their smaller parent line. Body weights, total erythrocyte counts, blood urea nitrogen (BUN) levels, and in vitro mitogenic response to PHA-P and Con A were higher in the F line birds. Line E had lower hemolytic complement levels, lower relative spleen and relative bursal weights, and a higher in vitro mitogenic response to PHA-P than its parent line. The Com B line had a lower toe web response to PHA-P, and lower serum levels of gamma-glutamyltransferase and bilirubin than Com A. Line Com B had higher total RBC counts and higher levels of alanine aminotransferase (ALT) than Com A. These results support the concept that some changes in the cell-mediated immune response, as well as other physiological changes that may potentially affect immune response, appear to accompany selection for faster growth.

Topics: Alanine Transaminase; Alkaline Phosphatase; Animals; Aspartate Aminotransferases; Blood Urea Nitrogen; Body Weight; Breeding; Cells, Cultured; Complement System Proteins; Concanavalin A; Eggs; Erythrocyte Count; Female; gamma-Glutamyltransferase; Growth; Hemolysis; Hypersensitivity; Leukocyte Count; Lymphocyte Activation; Lymphocyte Count; Lymphocytes; Male; Organ Size; Oviposition; Phytohemagglutinins; Spleen; Turkeys

The immunogenicity of a pore-forming polypeptide, equinatoxin II, from the sea anemone Actinia equina was studied after attenuation of the toxin's lethal and cytolytic activity by autologous polar lipids. In BALB/c mice, the lipid-inactivated toxin was used to raise specific antibodies and cellular immunity, resulting in in vivo protection. In vitro, haemolytic activity could be diminished by both normal and immune serum, the latter being more efficient. Purified specific IgG1 and IgG2 did not or only poorly neutralized the haemolytic activity, therefore implying the marked role of serum lipoproteins in the toxin attenuation. In response at the cellular level, equinatoxin II activated specific splenocytes. Increased concanavalin A stimulation of specific splenocytes was observed in the absence of antigen.

Topics: Animals; Antibodies; Antibody Specificity; Cnidarian Venoms; Concanavalin A; Dose-Response Relationship, Drug; Hemolysis; Immunization; Immunoglobulin G; Lipids; Mice; Mice, Inbred BALB C; Sea Anemones

The antibiotic bleomycin was examined as a possible component of hybrid molecules composed of an address fragment and a generator of reactive oxygen species. The bleomycin-Fe(II) complex was found to destroy the erythrocyte membrane by generating reactive oxygen. The ability of antioxidants to slow down haemolysis points to a free-radical mechanism for this process. The protective effects of catalase and superoxide dismutase indicate that hydrogen peroxide and the superoxide radical formed on autoxidation of the complex are essential for membrane damage. Haemolytic activity is also exhibited by bleomycin-Fe(III) reduced in the NADPH-cytochrome P450 reductase reaction. The covalent binding of bleomycin to such address molecules as concanavalin A and antierythrocyte immunoglobulin G enhances the ability of the bleomycin-Fe(II) complex to destroy the plasma membrane of erythrocytes.

Topics: Animals; Antioxidants; Bleomycin; Catalase; Concanavalin A; Erythrocyte Membrane; Hemolysis; Immunoglobulin G; Kinetics; Mice; Mice, Inbred CBA; Protein Binding; Superoxide Dismutase

The temperature (0 degrees C and 37 degrees C) and the medium tonicity (0.15-1.20 M NaCl) were shown to affect erythrocyte agglutination by concanavalin A. Treatment of cells with lectin caused no significant decrease in the erythrocyte hemolysis upon cooling. Diamide, unlike concanavalin A used at concentrations above 2.0 M decreases the cell sensitivity to the cold shock. The changes in the erythrocyte susceptibility to cooling within the temperature range of 37-0 degrees C correlate with changes in the electrophoretic spectrum of membrane proteins. The progressive decrease in the spectrin bands intensity with a simultaneous formation of high molecular weight protein aggregates not included in the gel composition was observed after diamide treatment. The diamide effect depends on the medium tonicity, at which the treatment was performed, being especially well pronounced in hypertonic media with 0.8-1.2 M NaCl concentrations, the maximal spectrin aggregation being observed under these conditions. It is suggested that the main factor of the mechanism underlying the erythrocyte hypertonic cold shock is the increase in the association of peripheral cytoskeleton proteins with plasma membrane in osmotically dehydrated cells which limits the ability of lipids to adapt during cooling and results in the stabilization of defects in the membrane structure at low temperatures. Diamide eliminates these unfavourable changes eventually resulting in the dissociation of peripheral proteins from the cytoplasmic surface of the membrane on the protein aggregation.

Topics: Adaptation, Physiological; Blood Proteins; Cold Temperature; Concanavalin A; Diamide; Erythrocyte Membrane; Erythrocytes; Hemolysis; Humans

Hybrid molecules consisting of an address peptide and an active oxygen-generating fragment may be used for selective destruction of cells. We tested the possibility of using the antibiotic bleomycin (BLM) as an active oxygen-generating fragment of such a molecule. It was found that bleomycin can induce destruction of cell membranes. BLM-mediated cell destruction was inhibited by addition of catalase, superoxide dismutase, and OH. scavangers (mannitol and ethanol), suggesting that hydroxy radical is involved in the process. BLM can induce membrane impairment at the expense of electrons supplied by NADPH-cytochrome P450 reductase. Covalent binding of BLM to an address fragment (concanavalin A, immunoglobulin G) increases the ability of BLM to destroy erythrocyte membranes. The data obtained lead to the conclusion that BLM can be used as an active oxygen-generating fragment of a proposed cell-destroying hybrid molecule.

Topics: Animals; Bleomycin; Catalase; Concanavalin A; Erythrocyte Membrane; Erythrocytes; Ethanol; Hemolysis; Immunoglobulin G; Kinetics; Mannitol; Mice; Mice, Inbred CBA; Potassium; Protein Binding; Superoxide Dismutase

Topics: Animals; Calcium; Complement Activation; Concanavalin A; Ethylamines; Hemolysis; In Vitro Techniques; Lipopolysaccharides; Magnesium; Mannose; Sheep; Sodium

Quantitatively much higher Concanavalin A (Con. A) agglutinability, haemolytic potency, and activities of acid hydrolases, namely phosphatase (EC 3.1.3.2), ribonuclease (EC 2.7.7.16), deoxyribonuclease (EC 3.1.4.5) and proteinase--were observed in a virulent strain of Entamoeba histolytica (IP-106), as compared to attenuated and avirulent strains (200-NIH) and DKB respectively. In addition, significant differences in these parameters were observed among clonal cultures derived from the latter two cultures by cultivation of single amoebic cells picked out by micromanipulation. Repeated sub-culturing of parent cultures of both these strains in cholesterol-enriched medium resulted in marked enhancement of all the above activities, but no such change occurred in the derived clonal cultures following similar cholesterol treatment. The implication of these findings in relation to enhancement of the virulence of E. histolytica by cholesterol is discussed.

Topics: Acid Phosphatase; Agglutination; Animals; Cholesterol; Concanavalin A; Culture Media; Deoxyribonucleases; Endopeptidases; Entamoeba histolytica; Hemolysis; Ribonucleases; Virulence

Crude extract prepared from isolated and purified nematocysts (stenoteles, desmonemes, isorhizas) of Hydra attenuata Pall. (Cnidaria, Hydrozoa) contains two main toxic proteins. The first is a hemolysin (100-200,000 mol.wt) which also causes initial spasmodic contractions in larval and adult specimens of Drosophila. Both, hemolytic and neurotoxic activities are inhibited by low concentrations of Triton X-100. The second protein (30-100,000 mol.wt), which is not susceptible to Triton causes long lasting paralysis leading to death of the test animals (LD50 approximately 5 mg crude nematocyst extract per kg). Neither of the toxins is identical with the previously described phospholipase.

Topics: Animals; Chromatography, Gel; Concanavalin A; Drosophila; Hemolysis; Humans; Hydra; In Vitro Techniques; Lethal Dose 50; Marine Toxins; Time Factors; Tissue Extracts

Repeated passage of the 200-NIH strain of Entamoeba histolytica through cholesterol-enriched axenic growth medium induced marked increases in cholesterol, phosphoglucomutase and hexokinase levels and a less prominent rise in the protein content of amoebic cells. There was also pronounced enhancement of haemolytic activity and Concanavalin A (Con A) agglutinability of the culture, but no significant change was observed in glucose phosphate isomerase. These cholesterol-induced effects persisted to a large extent when amoebae were subsequently repassaged through normal axenic medium lacking exogenous cholesterol, but changes in cellular cholesterol and protein levels did not persist. Qualitatively similar results were obtained whether the sterol was layered as a film on the glass walls of the culture tubes or supplied as sonicated micells, but the latter was in general more effective.

Topics: Agglutination; Animals; Cholesterol; Concanavalin A; Entamoeba histolytica; Glucose-6-Phosphate; Glucose-6-Phosphate Isomerase; Glucosephosphates; Hemolysis; Hexokinase; Phosphoglucomutase

The correlations among the potentiating activity of various PS analogs on concanavalin A (Con A)-induced rat mast cell degranulation, the hemolytic activity and the incorporation into the mast cell membrane were studied. The following results were obtained. Lysophosphatidylserine (LysoPS) caused rat mast cell activation (degranulation) in the presence of Con A. The order of the activity was as follows: 1-stearoyl lysoPS = 1-palmitoyl lysoPS greater than 1-myristoyl lysoPS greater than 1-lauroyl lysoPS. The relative hemolytic activity of these compounds was similar to that observed in the mast cell activation. Dilauroyl PS, which shows similar hemolytic activity to 1-myristoyl lysoPS, did not activate mast cells appreciably. The relative activity of these phospholipids in the binding to mast cells was 1-stearoyl lysoPS greater than dilauroyl PS greater than 1-lauroyl lysoPS. Hemolytic activity, as well as activity on mast cells, of lysoPS analogs was well correlated to mast cell membrane incorporation, whereas such a correlation was not found with PS analogs. Dilauroyl PS could be accumulated in the mast cell membrane and showed hemolytic activity, but did not activate histamine secretion.

Topics: Animals; Cell Membrane; Concanavalin A; Hemolysis; Histamine Release; Lysophospholipids; Male; Mast Cells; Phosphatidylserines; Rats; Rats, Inbred Strains; Serotonin

The role of mannose containing molecules in Sindbis virus envelope glycoproteins and cell membrane receptors was investigated by means of concanavalin A (con A). Treatment of virus or BHK 21 cells with the lectin before infection reduced viral infectivity. Hemagglutination and hemolysis showed a different sensitivity to con A. Hemagglutination was not affected by the lectin and treatment of erythrocytes with con A produced an enhancement of agglutinability by the virus. Hemolytic activity was, on the contrary, strongly reduced by the lectin. The inhibition observed is due to an action on viral structures and not to an interaction with erythrocyte receptors because con A did not affect the hemolysis when preincubated with erythrocytes.

Topics: Animals; Cell Line; Concanavalin A; Cricetinae; Erythrocytes; Geese; Hemagglutination, Viral; Hemolysis; Mannose; Receptors, Virus; Sindbis Virus; Viral Envelope Proteins; Virus Replication

Red blood cells were collected from Landrace X Duroc pigs in pooled and single batches. The RBC were stored for 24 hours to 20 days and exposed to 1 or more chemical and physical stressors. The chemicals were pyruvate, lactate, inosine, concanavalin A-luminol-bovine serum albumin conjugate, hydrogen peroxide, L-mimosine, and 3-amino-L-tyrosine. Physical stressors included thermogenic microwave radiation (2,450 MHz, mean specific absorption rate of 91 W/kg) and conventional heating with hot air or hot-water bath. Heating erythrocytes to 43 C for 10 minutes with microwaves or hot air did not significantly (P greater than 0.05) increase hemolysis, compared with hemolysis of RBC at 4 C (controls). Pyruvate or lactate did not affect the RBC under these conditions. Hemolysis of cells coated with concanavalin A-luminol-bovine serum albumin and heated to 43 C for 10 minutes by use of microwaves or hot air was significantly (P less than 0.05 by Student's t test) greater (48%) than that of RBC at 4 C (controls). The method of heating or the presence of pyruvate, lactate, or inosine did not have a significant (P greater than 0.05) effect on hemolysis. Lysis of RBC (14 days after collection) coated with concanavalin A-luminol-bovine serum albumin and stored at 4 C was not significantly different (P greater than 0.05) than that of noncoated cells (6 days after collection) stored at 4 C (control). When RBC were heated 20 days after collection to 48 C for 30 minutes, using a hot water bath, hemolysis was 60.6% greater than that of control cells (4 C).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Concanavalin A; Erythrocyte Aging; Erythrocytes; Hemolysis; Hot Temperature; Hydrogen Peroxide; Inosine; Lactates; Luminescent Measurements; Luminol; Mimosine; Pyruvates; Serum Albumin, Bovine; Swine; Time Factors; Tyrosine

The ability of concanavalin A (Con A) to inhibit the complement consumption by sheep erythrocytes coated with IgG, (IgG2-SpA1)2 complex and IgM anti-Forssman antibodies was studied. The (IgG2-SpA1)2 anti-Forssman antibody complex showed an increased hemolytic activity compared to uncomplexed IgG antibody. Con A in the presence of the (IgG2-SpA1)2 complex or the IgM antibody exhibited a high inhibitory capacity. An inhibitory capacity of Con A on IgG antibody hemolytic activity was also proved but at a Con A:IgG antibody concn ratio about 10 times higher than that for the Con A-IgM antibody system. A time dependence of Con A inhibitory capacity was determined, thus explaining the variable behaviour of the IgG antibody in the inhibition experiments. Radioactivity experiments demonstrated that only half of the Con A interacted with the (IgG2-SpA1)2 complex compared to the amount of Con A which interacted with IgG uncomplexed antibody.

Topics: Animals; Antigens, Heterophile; Complement System Proteins; Concanavalin A; Dose-Response Relationship, Immunologic; Erythrocytes; Forssman Antigen; Hemolysis; Immunoglobulin G; Immunoglobulin M; Sheep; Staphylococcal Protein A; Time Factors

Succinylated concanavalin A (SCon A) lyses sheep erythrocytes (E) in the presence of complement, whereas the native tetravalent lectin, Con A, is inactive. We have studied the ability of E-SCon A (ES) to interact with early acting guinea pig (gp) or human (hu) complement components (C1, C2, C4) and found that cell intermediates ESC1, ESC4, ESC14, and ESC142 can be generated that are analogous to intermediates conventionally prepared with E and rabbit IgM (pentameric) anti-Forssman antibody. Titration of gp or hu C1, C4, and C2, and quantification of the number of activated C1 molecules bound to ESC4 by the C1 fixation and transfer test showed in each case that an average of one effective site per cell was sufficient to cause cell lysis. Determination of tmax for optimal formation of ESC142 sites depended on the species combination of components used to make the intermediates, and the decay of ESC142 and EAC142 sites or sites generated with ESC4, EAC4, and trypsin-activated C2 were similar. The sugar alpha-D-methylglucopyranoside (alpha-MGP) inhibited binding of SCon A to E and eluted the lectin from ES, whereas galactose was nearly inactive, consistent with lectin sugar-binding selectivity. In contrast, both sugars were ineffective in eluting SCon A or C4hu from ESC4hu, indicating that C4hu blocked the interaction between lectin and alpha-MGP, perhaps by steric hindrance. SCon A is a divalent functional analogue of IgM anti-Forssman antibody that may be a uniquely suited reagent specific for cell membrane glycoconjugates for studying the mechanism of binding and activation of complement components.

Topics: Animals; Binding Sites, Antibody; Complement Activation; Complement C1; Complement C2; Complement C4; Complement Fixation Tests; Concanavalin A; Erythrocytes; Guinea Pigs; Hemolysis; Humans; In Vitro Techniques; Methylglucosides; Rats

Alpha toxin purified from Staphylococcus aureus strain Wood 46 and radioiodinated by the lactoperoxidase method retained full haemolytic activity and was used to study factors affecting binding to rabbit and horse erythrocytes. A relatively fixed percentage of added toxin bound to both cell types; the percentage bound was independent of temperature, pH, cell concentration and toxin concentration. Neither a 50-fold excess of native toxin nor Concanavalin A inhibited the binding of iodinated toxin to erythrocytes. The results suggest that differences in the sensitivity of erythrocytes to haemolysis do not reflect the abundance of high affinity toxin receptors on sensitive cells, but are more probably the result of differences in the intrinsic stability of the membrane and its sensitivity to perturbation by amphiphilic agents.

Topics: Animals; Bacterial Toxins; Concanavalin A; Erythrocytes; Hemolysin Proteins; Hemolysis; Horses; Hydrogen-Ion Concentration; Kinetics; Rabbits; Staphylococcus aureus; Temperature

A new test modification for the detection of stimulant-induced cytotoxicity of human mononuclear blood cells is described. Papain-treated human erythrocytes were used as indicator cells. The effector cells were lymphocytes. The ratio of target cells to effector cells was 10/1. Haemoglobin as a marker of lysis of the erythrocytes, released in the supernatant, was measured quantitatively in form of its pseudoperoxidase-activity. PHA, ConA and tannic acid were ascertained and tested as stimulants of cytotoxicity. The reaction was inhibitable by anti-human-lymphocyte-globulin. The test conditions were optimized in regard to incubation time, -temperature, -vessels, culture medium and target cells. The technique is easy to manipulate, has only slight pretensions to the equipment of the laboratory and appears to be very effective. We recommend to apply this method of stimulant-induced cytotoxicity within the detection of the immune state, especially in the progress of immunopathological diseases and the analysis of efficiency of immunosuppressive therapy.

Topics: Cells, Cultured; Concanavalin A; Cytotoxicity, Immunologic; Erythrocytes; Hemolysis; Humans; Lymphocytes; Phytohemagglutinins; Tannins

Mouse seminal plasma (SP), a mixture of aqueous extracts of prostate, seminal vesicle, and epididymis, exerts potent immunosuppressive effects in vivo. The SP mixture completely suppressed both primary and secondary humoral immune responses in mice to low immunizing doses of antigen (bovine serum albumin or washed epididymal sperm), and also significantly suppressed the antibody response to high doses of immunizing antigen. The humoral immune response to epididymal sperm was also suppressed when sperm were incubated in SP and then washed before immunization and when SP was administered at a secondary site (i. p.) at the time of sperm immunization (subcutaneous). When SP components were tested individually in in vitro assays, all of them (prostate, seminal vesicle, and epididymis) suppressed mitogen-induced lymphocyte transformation. Prostatic fluid also inhibited complement-mediated hemolysis in a standard immune hemolytic assay. These data indicate that potent immunosuppressive factors are present at several locations within the male reproductive tract; these factors may serve to protect sperm from immunologic damage and prevent sensitization of females to sperm antigens after insemination.

Topics: Animals; Antigens; Cattle; Complement System Proteins; Concanavalin A; Female; Hemolysis; Immunosuppression Therapy; Lymphocyte Activation; Male; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Phytohemagglutinins; Semen; Serum Albumin, Bovine

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

Native tetravalent Con A and the divalent acetylated derivative increased the hemolytic titer (i.e., the reciprocal of the antibody dilution required to give an average of one lytic site per sheep erythrocyte) of IgG antibodies against Forssman antigen by up to 225% with guinea pig and human complement. Although the average number of lytic sites generated at each antibody concentration increased, the slope of the titration curve did not change. Other lectins with the same or different sugar specificity either augmented or inhibited hemolysis but were less potent than Con A. Augmentation by Con A was consistent with the ability of lectin on the cell surface to bind but not activate guinea pig C1. Thus it appears that cell-bound Con A and IgG yield a complex that behaves like a doublet of IgG antibody molecules in its ability to fix and activate C1, when activation is dependent on the IgG component. In contrast, the highest dose of Con A inhibited by at least 50% the hemolytic activity of IgG antibodies against either a sugar-free protein (HSA) or a protein reactive with Con A (human myeloma IgE) using cells to which these antigens were coupled with chromic chloride. This suggests that the identity, density, and/or mode of presentation of the antigen on the cell surface may be important determinants of lectin-induced augmentation. Although both the enhancement or inhibition by Con A in the presence of whole C correlated with the number of C1 molecules bound and activated, there was no correlation with the ability of the lectin to agglutinate the cells.

Topics: Antigen-Antibody Complex; Complement C1; Concanavalin A; Forssman Antigen; Hemolysis; Lectins; Temperature

Topics: Animals; Complement Activation; Complement C1; Complement Fixation Tests; Complement Pathway, Classical; Concanavalin A; Dose-Response Relationship, Drug; Erythrocytes; Guinea Pigs; Hemolysis; Lectins; Methylglucosides; Sheep; Wheat Germ Agglutinins

Mouse lymph node cells sensitized with PHA or Con A in liquid phase grew into T-cell colonies when seeded in a two-layer soft agar culture system containing the mitogen. The colony cells were of T-cell lineage. This was deduced from their morphology, ultrastructure, positive strain for theta-isoantigen and the fact that no colonies were formed by lymphoid cells from congenitally athymic nude mice. The architecture of the colonies and their component cells was studied by scanning electron microscopy. Clonogenic assay indicated that macrophages are active modulators of T cell proliferation. Colony formation was markedly enhanced by hemolysate and/or amino acid, L-glutamine or L-cystine, added to the culture medium. The largest number of colonies grew when both the liquid and soft agar media were supplemented with hemolysate and one of the amino acids. Under these conditions the minimal seeding level for colony formation could be reduced from 2.0 X 10(5) to 1.6 X 10(4) cells/culture.

Topics: Animals; Cell Division; Concanavalin A; Cystine; Glutamine; Hemolysis; Lymphocyte Activation; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Inbred ICR; Mice, Nude; Mitogens; Phytohemagglutinins; Rats; T-Lymphocytes

Topics: Animals; Carbohydrates; Complement Activation; Complement Pathway, Classical; Concanavalin A; Dose-Response Relationship, Drug; Guinea Pigs; Hemolysis; Humans

A purified toga-alphavirus, Getah (GET), showed optimal hemolytic activity for one-day-old chick red blood cells when incubated at 37 C for 120 min at pH 6.2. Experimental data obtained from various angles, such as pH dependency, inhibition by virus-specific antiserum and by concanavalin A, indicated that the hemolysis was a property of the virus particle itself. Although the mechanism of hemolysis by togaviruses has not been known, our results indicated that viral lipids may participate in this activity since the hemolytic activity was impaired by delipidation procedures.

Topics: Animals; Arboviruses; Calcium Chloride; Chickens; Concanavalin A; Detergents; Erythrocytes; Hemolysis; Hydrogen-Ion Concentration; Immune Sera; Mice; Pancreatin; Solvents; Temperature

Topics: Concanavalin A; Dithiothreitol; Glycoproteins; Hemagglutination, Viral; Hemolysis; Parainfluenza Virus 1, Human; Viral Proteins

Topics: Antibodies; Antibodies, Viral; Antigens; Antigens, Viral; Concanavalin A; Erythrocyte Membrane; Erythrocytes; Fluorescent Antibody Technique; Hemolysis; Lectins; Parainfluenza Virus 1, Human

Topics: Concanavalin A; Erythrocyte Membrane; Erythrocytes; Hemolysis; Immune Sera; Parainfluenza Virus 1, Human; Receptors, Concanavalin A

The relationship between the structures of six native dextrans and their effects on nonspecific resistance to infection (n.s.r.i.) in mice and also anticomplementary activity has been studied. The data obtained showed that the n.s.r.i. activity of dextrans generally increased with increase of extent of branching, but no direct correlation between these two factors was found. Data on exodextranase-catalyzed hydrolysis of dextrans suggest that the length of the outer chains may be important for the n.s.r.i. activity of the dextrans. Dextrans characterized by a significant extent of branching were anticomplementary, but no relationship between extent of branching and anticomplementary activity was observed.

Topics: Animals; Anti-Bacterial Agents; Chemical Phenomena; Chemistry; Complement Fixation Tests; Complement Inactivator Proteins; Concanavalin A; Dextrans; Escherichia coli Infections; Glycoside Hydrolases; Guinea Pigs; Hemolysis; Male; Mice; Structure-Activity Relationship

Topics: Adsorption; Animals; Cats; Concanavalin A; Erythrocyte Membrane; Erythrocytes; Guinea Pigs; Hemagglutination, Viral; Hemolysis; Horses; Humans; In Vitro Techniques; Paramyxoviridae

Freeze-etch studies reveal that mild pronase treatment with subsequent incubation in concanavalin A induces aggregation of intramembranous particles (IMP) in intact human erythrocytes. This alteration in particle distribution is accompanied by a change in the distribution of the Con A molecules such that they also become clustered on the extracellular etch face. If divalent succinyl Con A is used after pronase instead of tetravalent Con A the IMPs still become clustered. Pronase only, Con A only, or succinyl Con A only does not cause the IMPs to become aggregated. Most surprising is the finding that pronase followed by Con A causes partial haemolysis of the cells whereas pronase only, Con A only, or pronase+succinyl Con A do not cause this haemoglobin loss. These perturbations of the erythrocyte plasma membrane appear to be a result of the pronase+Con A exerting a transmembrane effect on the spectrin. This conclusion is supported by sodium dodecylsulphate polyacrylamide gel electrophoresis of material crosslinked with dimethyl adipimidate dihydrochloride, which indicates that spectrin is more susceptible to being cross-linked after pronase+Con A; i.e. the spectrin is probably aggregated by the enzyme and lectin incubation.

Topics: Cell Aggregation; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Erythrocyte Membrane; Erythrocytes; Freeze Etching; Hemolysis; Humans; Membrane Proteins; Microscopy, Electron, Scanning; Pronase

Topics: Animals; Antibodies; Cattle; Concanavalin A; Eye; gamma-Globulins; Hemolysis; Immunity; Inflammation; Rabbits

When concanavalin A (1 microgram/ml) or wheat germ agglutinin (2 microgram/ml) was preincubated with a suspension of 2% rabbit erythrocytes for 5 min at 20 C, the binding [125I]-labeled staphylococcal alpha toxin to these erythrocytes was greatly inhibited and the hemolytic action of alpha toxin was decreased. The inhibitory effect of concanavalin A on hemolysis by alpha toxin was completely reversed in the presence of 0.1 M alpha-methyl-D-glucoside or alpha-methyl-D-mannoside. Phytohemagglutinin-P from Phaseolus vulgaris and soybean agglutinin inhibited hemolysis by the toxin at concentrations exceeding 20 microgram/ml. The effect of concanavalin A on alpha-toxin hemolysis was studied further to ascertain the nature of the inhibition. Double reciprocal plots were made of hemolysis against alpha toxin concentrations, and the data suggested that inhibition of the initial rate of the hemolysis by concanavalin A is competitive in nature. This was probably due to an interaction with the alpha toxin binding sites on the cell membrane surface.

Topics: Animals; Bacterial Toxins; Binding Sites; Binding, Competitive; Concanavalin A; Erythrocytes; Hemolysis; Kinetics; Lectins; Rabbits; Staphylococcus aureus

Lysis of sheep erythrocytes (E) sensitized with anti-Forssman antiserum (EA) is inhibited by the action of concanavalin A (Con A) on whole guinea pig complement (GPC). The degree of inhibition observed for a given quantity of GPC was dependent on the Con A concentration. Specifically, Con A inhibits the activity of the early acting complement components C1 and C2 in the fluid phase, but has no significant effect on lysis once these components are bound to EA. Results of tmax experiments performed in the presence or absence of Con A showed that inhibition of C2 activity results from a direct interaction between Con A and C2 and not from a decreased number of effective EAC14 sites. Furthermore, since Con A pretreated or untreated EAC14 cells had the same tmax value, Con A and C2 apparently do not compete for the same binding site on the indicator cells. The lectin has no observable effect on either fluid phase or cell-bound C4 activity. Under similar conditions, wheat germ or soy bean agglutinin, leucoagglutinin or pokeweed mitogen did not inhibit hemolysis.

Topics: Antibodies; Complement C1; Complement C2; Complement C4; Complement System Proteins; Concanavalin A; Erythrocytes; Hemolysis; In Vitro Techniques; Lectins

Concanavalin A inhibits serum 5'-nucleotidase activity, without causing significant inhibition of alkaline phosphatase activity. This observation serves as the basis for a new method for assaying the 5'-nucleotidase activity in serum, which depends upon the difference between the enzymic hydrolysis of adenosine-5'-monophosphate in the presence and absence of concanavalin A. A denosine released by the 5'-nucleotidase reaction is deaminated by a coupled reaction with adenosine deaminase to liberate inosine and ammonia, and ammonia is measured colorimetrically by the Berthelot reaction. In sera from 40 healthy adult persons, 5'-nucleotidase activity averaged 6.4 U/liter (SD, +/-2.0; range, 3-12). In sera from 100 patients, measurements of 5'-nucleotidase activity by the new assay averaged 8% lower than by a generally accepted method in which phenyl phosphate is used to suppress hydrolysis of adenosine-5'-monophosphate by alkaline phosphatase activity. The clinical validy of the new assay was tested by measuring serum 5'-nucleotidase activities in rats with bile duct ligation and in rats treated with thioacetamide to induce hepatocellular injury.

Topics: Adult; Aged; Concanavalin A; Female; Hemolysis; Humans; Indicators and Reagents; Kinetics; Magnesium; Male; Methods; Middle Aged; Nucleotidases; Spectrophotometry

A number of T-independent antigens and B cell mitogens were examined for their ability to activate C3 via the alternative pathway of the complement system. Loss of hemolytically active C3, generation of anaphylatoxin activity, and immunoelectrophoretic conversion of C3 and factor B, were checked in normal and C4-deficient guinea pig serum, and, in some cases, in normal human serum. As judged by their activity in these assays, 10 lipopolysaccharides of different origin and constitution, pneumococcus type III polysaccharide, levan, dinitrophenylated aminoethyl-dextran, dinitrophenylated (D-glutamic acid, D-lysin) copolymer, polymerized flagellin, and pokeweed mitogen were all capable of initiating the alternative pathway, but differed with respect to their potency, their relative activity in the presence or absence of C4, and their ability to inhibit C3-turnover at high concentrations. Polyvinylpyrrolidone of intermediate molecular weight (4 x 10(4) daltons) was only active if the most sensitive assay was used (anaphylatoxin generation). Other species of polyvinylpyrrolidone, depolymerized pneumococcal polysaccharide, aminoethyl-dextran, [D-glutamic acid, D-lysin] copolymer, phytohemagglutinin and concanavalin A failed to activate C3. C3-consumption by concanavalin A was due to nonspecific binding.

Topics: Anaphylaxis; Antigens, Bacterial; B-Lymphocytes; Complement C3; Complement C4; Complement System Proteins; Concanavalin A; Dextrans; Dinitrophenols; Escherichia coli; Flagellin; Fructose; Hemolysis; Ileum; Immunoelectrophoresis; Lectins; Lipopolysaccharides; Polysaccharides, Bacterial; Povidone; Salmonella; Streptococcus pneumoniae; T-Lymphocytes; Veillonella

Topics: Chromatography, Affinity; Complement C1; Complement C1 Inactivator Proteins; Complement Inactivator Proteins; Concanavalin A; Deoxyribonucleases; Hemolysis; Humans; Immunodiffusion; Proteins

Topics: Binding Sites; Complement C1; Complement C2; Complement Inactivator Proteins; Concanavalin A; Hemolysis

Topics: Animals; Chromatography, Gel; Cnidaria; Concanavalin A; Drug Interactions; Erythrocytes; Female; Freezing; Hemolysis; In Vitro Techniques; Lethal Dose 50; Mice; Mice, Inbred ICR; Sheep; Toxins, Biological

Topics: Animals; Binding Sites; Cell Membrane; Concanavalin A; Erythrocytes; Hemagglutinins, Viral; Hemolysis; Horses; Monosaccharides; Parainfluenza Virus 1, Human

Topics: Amanitins; Amino Acids; Animals; Chromatography, DEAE-Cellulose; Chromatography, Gel; Concanavalin A; Glycoproteins; Hemolysis; Isoelectric Focusing; Lethal Dose 50; Mice; Molecular Weight; Rabbits; Rats

Topics: Alkylation; Animals; Antibody-Producing Cells; Centrifugation, Density Gradient; Chickens; Complement System Proteins; Concanavalin A; Coombs Test; Hemolysis; Hemolytic Plaque Technique; Immune Sera; Immunoglobulin G; Immunoglobulin M; Rabbits

Topics: ABO Blood-Group System; Antigen-Antibody Reactions; Blood Proteins; Cell Membrane; Chromatography, Affinity; Concanavalin A; Erythrocytes; Hemolysis; Humans; Immunoglobulin Fab Fragments; Immunoglobulin G; Lectins; Microscopy, Electron; Models, Biological; Molecular Conformation; Neuraminic Acids; Neuraminidase; Vibrio cholerae

Topics: Animals; Antimycin A; Concanavalin A; Cytotoxicity Tests, Immunologic; Erythrocytes; Freund's Adjuvant; Guinea Pigs; Hemolysis; Immunoelectrophoresis; Immunoglobulin G; Lymphocytes; Male; Spleen; Thyroglobulin; Thyroiditis