concanavalin-a and Anaphylaxis

Nowadays there is growing evidence that some cytokines regulate biological functions of the mature mast cells. Therefore, we have studied whether TNF-alpha, the cytokine of multifunctional activities, could directly stimulate rat peritoneal mast cells to histamine secretion and whether it could modulate rat mast cell reactivity in anaphylactic (with ConA) and anaphylactoid (with compound 48/80) reactions. We have established that rat recombinant TNF-alpha does not activate rat mast cells to histamine release. However, TNF-alpha-treatment causes the decrease of spontaneous histamine release up to 85% (TNF-alpha concentration: 2 x 10(-9) M). Pretreatment of mast cells with TNF-alpha inhibits ConA-stimulate release of histamine with the percent release decreasing up to 33.7% of the control value (TNF-alpha concentration: 5 x 10(-9) M) and this decrease is statistically significant. Pretreatment of mast cells with TNF-alpha reduces compound 48/80-dependent histamine release as well and the percent release of histamine fell to 64.7% of the control value. We have concluded that TNF-alpha may play a significant role in regulation of mast cell secretory activity.

Topics: Anaphylaxis; Animals; Concanavalin A; Female; Histamine Release; Mast Cells; Rats; Rats, Wistar; Recombinant Proteins; Tumor Necrosis Factor-alpha

Mast cells represent significant cellular element of the skin. It is now postulated that they play an important role in cutaneous homeostasis and are engaged in some pathological processes as well. The aim of our study was to examine sensitivity of human cutaneous mast cells to anaphylactic and anaphylactoid stimuli. The studies were performed on human cutaneous mast cells obtained from healthy skin by enzymatic dispersion technique. The mast cells were activated in vitro with anti-IgE, concanavalin A (Con A), compound 48/80, substance P (SP) or tumor necrosis factor alpha (TNF-alpha). We have noticed that skin mast cells were susceptible to the challenge with anti-IgE and Con A, and histamine release was dose- and time-dependent. In both cases histamine release was high (44.0 +/- 4.1% with anti-IgE at dilution 1:500 and 20.1 +/- 2.4% with Con A in concentration 500 micrograms/ml). Cutaneous mast cells were challenged in a dose- and time-related fashion with compound 48/80, however histamine release was low (9.8 +/- 2.4%, at concentration of compound 48/80-100 micrograms/ml). SP and TNF-alpha also activated mast cells but the magnitude of histamine release was not high (up to 7.1 +/- 0.9%, SP in concentration 10(-4) M and 17.4 +/- 1.1%, TNF-alpha in concentration 10(-6) M) and maximal after 20 min reaction.

Topics: Anaphylaxis; Antibodies, Anti-Idiotypic; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Immunologic; Histamine Release; Humans; Immunoglobulin E; Kinetics; Mast Cells; p-Methoxy-N-methylphenethylamine; Sensitivity and Specificity; Skin; Substance P; Tumor Necrosis Factor-alpha

The study was undertaken to examine the effect of isoprinosine treatment on in vitro histamine release from peritoneal mast cells. The experiments were done on mice which received isoprinosine orally, at a dose of 50 or 100 mg per kilogram of body weight per day, in divided doses, every 8 hours, for a period of 1-9 days. After isoprinosine treatment, there was a significant decrease of in vitro Con A-induced histamine release from mast cells. This inhibitory effect was observed in both tested groups. In the second experiment, with mice sensitized with egg albumin, the treatment with isoprinosine gave also a significant inhibition of anaphylactic histamine release from mast cells, as compared with that of control without isoprinosine treatment. Results of both experiments suggested that isoprinosine may have some inhibitory effect on mast cell activation.

Topics: Anaphylaxis; Animals; Concanavalin A; Depression, Chemical; Female; Histamine Release; Inosine Pranobex; Mast Cells; Mice; Mice, Inbred BALB C; Peritoneal Cavity

Experimentally-induced type 1 hypersensitivities were induced in normal dogs to either ovalbumin or Ascaris antigen. In vitro and in vivo cell-mediated immune responses were measured before sensitization and again at 1 and 6 days after induction of anaphylaxis by intravenous challenge with antigen. Histamine-modulated lymphocyte functions, such as histamine-induced suppression, histamine co-mitogen induced blastogenesis and the in vivo cutaneous responses to intradermally injected mitogens decreased post anaphylaxis. Spontaneous suppression of the autologous mixed-lymphocyte reaction increased post anaphylaxis. Lymphocyte blastogenic response to Concanavalin A (Con A) decreased at 6 (but not at 1) days post anaphylaxis probably due to a mediator other than histamine. Blastogenesis of 24 h preincubated cells by suboptimal concentration of Con A, declined post anaphylaxis, but Con A-induced suppression was not significantly altered. Dogs with atopic dermatitis have some altered cell-mediated immune responses. Altered histamine-induced and spontaneous suppression, histamine suppression of mitogenesis and decreased contact sensitivity observed in this experimental type 1 hypersensitivity mimicked that of atopic dogs. Increased cutaneous response to mitogens observed in atopic dogs was not reproduced in the type 1 hypersensitive dogs. These findings suggest some of the altered cell-mediated immune functions observed in dogs with atopic dermatitis result from type 1 hypersensitivity. The other abnormalities may be intrinsic to the atopic state.

Topics: Anaphylaxis; Animals; Antigens, Helminth; Ascaris; Concanavalin A; Dog Diseases; Dogs; Drug Synergism; Female; Histamine; Immune Tolerance; Immunity, Cellular; Immunization; Intradermal Tests; Lymphocyte Activation; Lymphocytes; Male; Ovalbumin

The effects of coumarins on anaphylactic mediator release from rat mast cells were investigated. Since Pd-Ia (3'-angeloyloxy-4'-acetoxy-3',4'-dihydroseselin) causes relaxation of smooth muscle by inhibiting calcium influx, and since mediator release is a calcium-dependent process, studies were made on whether coumarins block calcium influx into rat mast cells stimulated by concanavalin A. Pd-Ia isolated from Peucedanum praeruptorum Dunn and related compounds, named Pd-C-II, Pd-C-III and Pd-C-IV, from Peucedanum decursivum Maxim., (Angelica decursiva Fr. et Sav.) inhibited anaphylactic mediator release from purified mast cells induced by concanavalin A with phosphatidylserine; their IC50 values were 79, 100, 102 and 73 microM, respectively. Pd-III, decursidin and water-soluble analogues of Pd-Ia (Pd-Ia-OH, Pd-Ia-OCH2CH3) did not inhibit the release. Thus the inhibitory actions of coumarins on calcium influx seemed to depend on the chemical structures of these compounds; an acetoxyl residue at position 3' or 4' on the seselin or xanthyletin skeleton might be essential for an inhibitory effect. Furthermore, Pd-Ia and Pd-C-III caused more than 40% reduction in 45Ca uptake induced by concanavalin A, whereas decursidin had little effect on it. Therefore, the inhibitions of mediator release of mast cells by some of coumarins from "Qian-Hu" seem to depend on their effects in blocking calcium influx.

Topics: Anaphylaxis; Animals; Calcium; Calcium Channel Blockers; China; Concanavalin A; Coumarins; Histamine Release; In Vitro Techniques; Male; Mast Cells; Plant Lectins; Plants, Medicinal; Rats; Rats, Inbred Strains

The peritoneal mast cells of rat (Wistar X August) F1 were incubated with purified mouse IgE antibodies and then challenged (in the presence of 50% D2O) with antigen, anti-mouse IgE and concanavalin A. It was found that incubation of mast cells with IgE antibodies led to different level of cell sensitization (in mast cells from different rats), expressed in antigen-induced histamine release (0-52%). Moreover, a) significant antigen-induced histamine release was usually accompanied by high Con A- and anti-IgE-induced histamine release from these cells; the magnitude of release was comparable to Con A- and anti-IgE-induced release from control, nonsensitized cells of the same rat; b) low antigen-induced histamine release was accompanied by the decrease of Con A- and anti-IgE-induced release, as compared to the release from control cells. This fall of reactivity to Con A and anti-IgE was statistically significant and was irreversible during 120 min.

Topics: Anaphylaxis; Animals; Concanavalin A; Histamine Release; Immunization, Passive; Immunoglobulin E; Male; Mast Cells; Rats; Rats, Inbred Strains

Mature circulating granulocytes have been considered functionally end stage cells unable to reconstitute their specific cytoplasmic granules. We have reevaluated this assumption by studying guinea pig peripheral blood basophils maintained in vitro for periods up to 72 hours after anaphylactic degranulation. Guinea pig basophils degranulated in vitro by exposure to either specific antigen (sheep serum) or lectin (Concanavalin A) synthesized new cytoplasmic granules. This process was characterized by the appearance of abundant rough endoplasmic reticulum, activation of the Golgi zone, interiorization of plasma membrane, and successive formation of empty vacuoles, multivesicular bodies, immature granules, and, finally, typical mature basophil granules. Although basophil neogranulogenesis in vitro was similar in most respects to that occurring in the bone marrow, it differed significantly in that regranulating basophils retained the nuclear characteristics of mature granulocytes. New basophil granule formation was more prominent and developed earlier when sheep serum was employed as the degranulation stimulus. Cultures degranulated with Concanavalin A, and, less commonly, sheep serum, also contained small numbers of basoblasts, large bizarre basophils with immature nuclear and cytoplasmic features, cytoplasmic lipid droplets, and mature, immature, and fused granules. These cells may arise by a process analogous to lymphocytes blast transformation.

Topics: Anaphylaxis; Animals; Antigens; Basophils; Concanavalin A; Guinea Pigs; Kinetics; Microscopy, Electron; Sheep; Vacuoles

Anaphylactic degranulation of guinea pig basophilic leukocytes, induced in vitro either with Concanavalin A or sheep serum (antigen), was resolved by transmission electron microscopy into two phases: (1) fusion of cytoplasmic granule membranes to form degranulation sacs communicating with the extracellular space by narrow pores and (2) resolution of degranulation sacs with concomitant granule matrix extrusion. Fusion of granule membranes occurred in the absence of obvious alterations of cytoplasmic filaments or microtubules but was preceded by a rapid increase in the number of 50- to 70-nm. cytoplasmic vesicles, a process evident 1 minute after exposure to lectin. By 5 minutes and at later intervals up to 20 minutes, as individual granule membranes fused to form degranulation sacs, vesicle frequency plunged to values one-half or less of control levels. Cytoplasmic vesicles were apparently incorporated into degranulation sacs and may have had a role in joining together the membranes of adjacent granules. Histamine release, detected at 5 minutes and maximal at 20 minutes, occurred at times when communications between degranulations sacs and the extracellular space were so narrow as to retain most recognizable granule matrix material. Resolution of degranulation sacs proceeded over a period of a day in culture and, in Concanavalin A-induced anaphylaxis, was sometimes incomplete even after 36 hours. During this phase, the frequency of cytoplasmic vesicles returned to normal or supernormal values, and the thin cytoplasmic processes forming the walls of degranulation sacs developed prominent, longitudinally disposed cytoplasmic filaments and ultimately retracted into the main cell body, depositing the membrane-free cytoplasmic granule matrix material outside the perimeter of the cell. Guinea pig basophil anaphylactic degranulation thus differs morphologically and kinetically from mast cell and basophil degranulation in other species in which granule membrane fusion and granule matrix extrusion occur nearly stimultaneously and are complete within minutes. The guinea pig basophil provides a useful model for dissociating these two intrinsic components of the degranulation process.

Topics: Anaphylaxis; Animals; Basophils; Concanavalin A; Cytoplasmic Granules; Exocytosis; Guinea Pigs; Histamine Release; Microscopy, Electron; Time Factors

Peritoneal mast cells from rats of BH- and DA-inbred strains were compared for their capacity to passive sensitization in vitro with mouse IgE antibodies. In identical experimental conditions mast cells of BH-rats were good receptors for mouse IgE anti-KLH fraction and released histamine when challenged with specific antigen, whereas mast cells of DA-rats were much less sensitized and released little histamine on the challenge. In contrast, mast cells of DA-rats wee more susceptible to the challenge with anti-rat IgE and Concanavalin A, than were cells of BH-rats. This susceptibility correlated with total serum IgE level, which was higher in DA- than in BH-rats. These results show higher concentrations of IgE on mast cells of DA--rats. It is suggested that rat IgE present on normal mast cells may be one of the factors interfering in in vitro sensitization of the cells from DA-rats with mouse IgE antibodies.

Topics: Anaphylaxis; Animals; Ascitic Fluid; Concanavalin A; Female; Histamine Release; Immunoglobulin E; Mast Cells; Mice; Rats; Rats, Inbred Strains

The relative activities of dextran, mannan and ovomucoid as agents inducing anaphylactoid responses in rats vary with the route of injection, mannan being the most active especially by the intradermal route. All of these responses are potentiated by pretreatment with insulin and inhibited by glucose. Carrageenan and concanavalin A are also active but only by the subcutaneous route. Whereas the concanavalin A response is potentiated by insulin, that of carrageenan is reduced. There are therefore at least three classes of agents capable of inducing anaphylactoid responses in rats.

Topics: Anaphylaxis; Animals; Carrageenan; Concanavalin A; Dextrans; Injections, Intradermal; Injections, Intraperitoneal; Injections, Subcutaneous; Insulin; Mannans; Ovomucin; Rats

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