concanavalin-a and Rhinitis--Allergic--Seasonal

The goal of our study was to examine spontaneous and stimulated apoptosis of peripheral blood MNC from allergic patients, sensitized to Der p I antigen as compared to cells from non-atopic subjects. Furthermore we aimed to investigate which populations of mononuclear cells (lymphocytes, monocytes) undergo the apoptosis and to determine relations between apoptosis and serum levels of sFas/APO-1, ICE/caspase-1 or TNF-alpha.. The study included 17 patients with perennial, allergic asthma and/or allergic rhinitis [6 male and 11 female; mean age 29,5 years; (range 15-49)]. Apoptosis was assessed by fluorescence technique and confirmed by flow-cytometric method and DNA ladder. Serum levels of sFas, ICE/caspase-1 or TNF-alpha were determined by immunoassays (ELISA).. Apoptotic index of unfractionated mononuclear cells (MNC) and lymphocytes (but not monocytes) were significantly higher in allergic patients as compared to non-allergic subjects after 48 and 72 hours of culture (p<0.05). Incubation of cells with ConA (10 microg/ml) resulted in a significant increase in the proportion of apoptotic cells in all populations once the apoptotic index for MNC and lymphocytes (but not monocytes) was again significantly higher in allergic as compared to non-allergic subjects after 24, 48 and 72 hour of culture. In allergic patients, mean serum sFas level, was significantly lower then in non-allergic group (mean value 624.8 pg/ml +/- 25.67 versus 802.0 pg/ml +/- 31.91; p = 0.003) and in both groups sFas level correlated inversely with apoptosis of MNC. The mean ICE/caspase-1 concentration was significantly higher in sera of allergic patients as compared to non-allergic group (mean value 27.71 pg/ml +/- 3.79 vs 23.54 pg/ml respectively; p<0.01). ICE/caspase-1 levels in allergic patients correlated with apoptotic index of mononuclear cells (r = 0.57; p<0.001).. An increased spontaneous and mitogen-induced apoptosis of MNC from peripheral blood of atopic patients as well as different serum levels of sFas and ICE/caspase-1 correlating with apoptosis, suggest different regulation of apoptotic process in peripheral blood mononuclear cells of patients with allergic asthma and/or rhinitis.

Topics: Adolescent; Adult; Apoptosis; Asthma; Caspase 1; Concanavalin A; fas Receptor; Female; Flow Cytometry; Humans; Leukocytes, Mononuclear; Male; Middle Aged; Rhinitis, Allergic, Seasonal; Tumor Necrosis Factor-alpha

Lymphocytes play a central regulatory role in mechanisms contributing to impaired function of immune system in atopy. The aim of our study was evaluate the mitogen-induced proliferation of lymphocytes in a group of asymptomatic, seasonal allergic rhinitis patients. A highly significant lower mitogen-induced proliferation and, in contrast to other studies, significantly lower background proliferative activity of lymphocytes were found in the atopic persons, comparing to the controls. We concluded that the decreased mitogen-induced proliferation of lymphocytes observed in allergic patients reflects abnormal T cell function, which is due to the atopic status, and not only as it was believed to the antigen-induced lymphocyte activation.

Topics: Adolescent; Adult; Concanavalin A; Female; Humans; Lymphocyte Activation; Male; Phytohemagglutinins; Rhinitis, Allergic, Seasonal

Loratadine, a new nonsedating histamine H1-antagonist, has been shown to inhibit immunologic release of inflammatory mediators in addition to its H1-receptor blocking properties. After oral administration, the agent is metabolized primarily to desethoxycarbonyl-loratadine (DCL). The basic piperidine, DCL, is readily soluble in water, whereas the nonbasic urethane, loratadine, is insufficiently soluble in water for some in vitro investigations. Therefore we used the metabolite, DCL, to study its influence on in vitro leukocyte histamine release (LHR) in 24 allergic and 22 nonallergic subjects. IgE-mediated and calcium ionophore A23187-induced LHR were inhibited by DCL in a dose-dependent fashion (values of drug concentration to induce 30% inhibition after stimulation with inhalant antigen, anti-IgE, concanavalin A, and calcium ionophore A23187 were 6, 8, 5, and 11 mumol/L, respectively). Higher concentrations of DCL caused mediator release in all subjects (n = 45, 30 mumol/L DC: 11% +/- 2% LHR, 100 mumol/L DCL: 35% +/- 1% LHR), abolishing any inhibitory effect of the drug. Rapid onset of inhibition by 10 mumol/L DCL was found in kinetic studies (n = 10). The inhibition of anti-IgE-induced histamine secretion was synergistically increased by simultaneous preincubation of DCL with the potent histamine H2-agonist, FRA-19. Additional data indicate that the inhibition of LHR by DCL might involve biochemical events that occur after cellular Ca++ influx because LHR induced by N-formyl-methionyl-leucyl-phenylalanine or the phorbol ester, 12-O-tetradecanoyl phorbol-12-acetate, was not significantly affected by DCL.

Topics: Basophils; Calcimycin; Concanavalin A; Histamine H1 Antagonists; Histamine Release; Humans; Immunoglobulin E; In Vitro Techniques; Loratadine; N-Formylmethionine Leucyl-Phenylalanine; Rhinitis, Allergic, Seasonal; Tetradecanoylphorbol Acetate

The binding of dog immunoglobulins G, A, M and E to concanavalin A (Con A) has been investigated. A passive cutaneous anaphylaxis test was used for measurement of dog IgE, and enzyme-linked immunosorbent assay was used for measurement of dog IgG, IgA and IgM. After the dog serum fraction was applied to a Con A-Sepharose column, sequential elution with different buffers was performed; 100% of IgE and IgM, 60% of IgG and 58% of IgA bound to the Con A-Sepharose. IgE was eluted by mannose, methylglucose, and methylmannoside. IgG was eluted by glucose, mannose, methylglucose, and methylmannoside. IgA and IgM were eluted by methylmannoside only. This provides a useful technique in the purification of dog immunoglobulins, especially dog IgE.

Topics: Animals; Chromatography, Affinity; Chromatography, Gel; Concanavalin A; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Immunoglobulins; Passive Cutaneous Anaphylaxis; Rhinitis, Allergic, Seasonal; Sensitivity and Specificity

Topics: Concanavalin A; Culture Media; Humans; Immunoglobulin E; In Vitro Techniques; Lymphocyte Activation; Phytohemagglutinins; Rhinitis, Allergic, Seasonal; T-Lymphocytes; Thymus Extracts

The helper/suppressor T cell ratio, as defined by monoclonal antibodies, was significantly higher in hay fever sufferers compared with controls (P less than 0.05), but only during or shortly after the pollen season. This was due to a reduction in the suppressor subset, which returned to control values in the winter. There was no significant difference in the non-specific concanavalin A-induced suppressor cell function compared with controls. The mean summer value was significantly lower than the winter value (P less than 0.05), but we cannot be sure that this was not the result of changes in laboratory conditions. No relationship was found between T cell subsets or suppressor cell function and total or specific IgE levels, or between T cell subsets and suppressor cell function. Our findings suggest that in hay fever, reduction in suppressor cell numbers and function is a secondary phenomenon.

Topics: Adolescent; Adult; Antibodies, Monoclonal; Concanavalin A; Female; Humans; Immunoglobulin E; Male; Middle Aged; Rhinitis, Allergic, Seasonal; Seasons; T-Lymphocytes, Regulatory

We have studied the influence of substance P (SP) on the proliferative response of concanavalin A (ConA)-activated peripheral blood lymphocytes from 16 birch pollen-allergic patients, sampled before and during the pollen season, and from 15 normal individuals. The median response to ConA 3 micrograms/ml in the presence of SP 10(-11)-10(-6) M, was in most instances within +/- 10% of the control value for cells from both healthy and atopic individuals. However, the individual differences were considerable. Analysis of the proliferative responses to ConA of the cells from the allergic patients sampled before and during season, revealed higher responses in the presence of 10(-6) than of 10(-7) M SP. This was in contrast to the findings in the normal individuals: only half of their cells showed such increased responses. This difference in response frequency was statistically significant between allergic patients before season and normal individuals (P less than 0.05) and between allergic patients during season and normal individuals (P less than 0.01). The difference in proliferation rate in the SP concentration interval, 10(-6) to 10(-7) M, for the cells from allergic patients, sampled both before and during season, was significantly different from the cells from healthy individuals (P less than 0.03 and P less than 0.001 respectively). The cells sampled from four allergic subjects during the birch pollen season showed a more profoundly decreased response to ConA in the presence of SP 10(-8) M, compared with their cells sampled before season. Such responses were never seen with cells sampled before season and with cells from normal individuals. The results suggest an involvement of SP in the immunoregulation, particularly in patients exhibiting allergic reactions to birch pollen.

Topics: Adult; Cells, Cultured; Concanavalin A; Dermatitis, Atopic; Humans; Lymphocyte Activation; Middle Aged; Plant Lectins; Pollen; Rhinitis, Allergic, Seasonal; Rosette Formation; Substance P

Histamine-induced suppressor cell function was measured in a group of twenty-five hay fever suffers and sixteen non-atopic controls. No difference was found in the suppression index between sufferers and controls. There was, however, a significant relationship between the suppression index and nasal sensitivity to grass pollen, whether measured pre- or post-season, but no correlation between specific or total IgE levels and the suppression index. Histamine-induced suppressor cell function may influence pollen sensitivity in the most sensitive subjects, but not via an effect on IgE production.

Topics: Adolescent; Adult; Concanavalin A; Female; Histamine; Humans; Immunoglobulin E; In Vitro Techniques; Lymphocyte Activation; Male; Nasal Provocation Tests; Plant Lectins; Pollen; Rhinitis, Allergic, Seasonal; T-Lymphocytes, Regulatory

It has been suggested that IgE-dependent basophil histamine release (HR) does not necessarily relate to the amount of cell-bound IgE and, therefore, basophil "releasability" must be considered an important factor in this secretory process. To compare an IgE-dependent basophil HR process in nonatopic subjects and patients with allergic rhinitis, concanavalin A (Con A) was used as a secretagogue to stimulate mediator secretion. In 1.0 mmol/L of calcium-containing buffer, basophil HR to Con A (3.0 mcg/ml) was 50.2 +/- 8.6% in patients with allergic rhinitis and only 10.1 +/- 3.9% in nonatopic subjects. To evaluate whether this enhanced HR might be related to increased membrane influx of calcium, the following strategy was followed. Strontium (3.0 and 10.0 mmol/L) enhances immunologic (IgE) release of basophil histamine. Although the mechanism for strontium enhancement is not established, strontium may pass through the membrane channel more easily than calcium to increase secretion. We reasoned that if the enhanced release of histamine to Con A was related to increased membrane permeability to calcium, stimulation of basophil histamine secretion in the presence of strontium would reduce this difference. In both nonatopic subjects and patients with allergic rhinitis, strontium (3.0 and 10.0 mmol/L) enhanced HR. Enhanced HR with strontium was greater with basophils from normal subjects than from subjects with allergic rhinitis. Whether our observations with strontium indicate that the enhanced histamine releasability to Con A in subjects with allergic rhinitis may, in part, be due to a greater influx of calcium after immunologic stimulation must await characterization of the strontium effect or direct measurements of calcium ion disposition.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Basophils; Buffers; Calcimycin; Concanavalin A; Dose-Response Relationship, Immunologic; Female; Histamine Release; Humans; Immunoglobulin E; Male; Rhinitis, Allergic, Seasonal; Strontium

Affinity-purified IgE-binding factors from the plasma of patients with the hyper IgE syndrome (HIE) were assessed for their capacity to enhance IgE synthesis by B cells derived from patients with allergic rhinitis or normal nonatopic donors. IgE-binding factors from three of four HIE patients enhanced IgE synthesis by B cells from patients with perennial allergic rhinitis, or with seasonal allergic rhinitis (SAR) and recent pollen exposure, but did not enhance IgE synthesis by B cells from nonatopic donors or from SAR patients with no recent pollen exposure. IgG synthesis was not affected by HIE IgE binding factors. In contrast, IgE binding factors from three of three nonatopic donors failed to enhance IgE or IgG synthesis. Plasma IgE-binding factors from the fourth patient with HIE contained a mixture of IgE-potentiating activity and IgE-suppressive activity. These two activities could be separated on concanavalin A Sepharose or peanut agglutinin agarose columns. Human IgE potentiating factor, but not IgE suppressive factor, had affinity for concanavalin A but not peanut agglutinin and fractionated into two peaks on gel filtration over Sephadex G-75: one peak with a molecular size of approximately 15,000 D and the other with a molecular size of approximately 60,000 D. The isolation of functional IgE binding factors which potentiate IgE synthesis from the plasma of patients with HIE suggests that IgE-binding factors play an important role in the in vivo regulation of IgE synthesis in man.

Topics: B-Lymphocytes; Concanavalin A; Humans; Hypergammaglobulinemia; Immunoglobulin E; Lectins; Lymphokines; Molecular Weight; Peanut Agglutinin; Prostatic Secretory Proteins; Rhinitis, Allergic, Seasonal

Spontaneous in vitro production of IgE was found higher in a group of untreated grass sensitive atopic patients than in normal volunteers when assessed at the cellular level with a reverse hemolytic plaque assay. This study also confirmed the increase of IgE synthesis after pokeweed mitogen stimulation in non-atopic donors and its decrease in atopic patients. Moreover, in this work we looked for a potential defect in immunoregulatory functions in atopic patients toward the in vitro IgE production. Indeed, histamine is known to activate suppressor cells capable, in turn, of suppressing IgG and IgE production from normal cells. In atopic patients, histamine could activate cells capable of suppressing IgG production but not IgE. Furthermore, similar findings were found when Concanavalin A-induced suppressor cells were examined. These findings suggest (a) a defective regulatory function towards IgE in atopic patients and (b) that the same subpopulation of suppressor cells seems to be activated by histamine and ConA and defective in atopic patients.

Topics: Antibody-Producing Cells; B-Lymphocytes; Concanavalin A; Hemolytic Plaque Technique; Histamine; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Immunoglobulin G; Pokeweed Mitogens; Rhinitis, Allergic, Seasonal; T-Lymphocytes, Regulatory

Topics: Animals; Basophils; Bronchi; Concanavalin A; Cyclic AMP; Dogs; Haplorhini; Histamine Release; Humans; Immune Sera; Immunoglobulin G; Iodine Radioisotopes; Isoproterenol; Macaca; Mast Cells; Plant Lectins; Pollen; Rabbits; Radioimmunoassay; Rhinitis, Allergic, Seasonal; Sheep; Theophylline