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Biomedical subjects

H J Wedner

Publications and source records attributed to H J Wedner.

At least 37 records · Page 2Linked to original sources
PubMed1988

Immunoblotting of pollen proteins: Tween 20 mediates nonspecific binding of gammaglobulins to immunoblots of oak-pollen proteins.

Oak-pollen extracts, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred onto nitrocellulose, demonstrated extensive banding in the absence of human sera or with nonatopic sera with both a horseradish peroxidase-conjugated goat antihuman IgE and 125I-labeled rabbit antihuman IgE. When the nonionic detergent Tween 20 was removed from all incubations and washes, negative banding appeared, corresponding to bands that were clearly visible when Tween 20 had been used as a blocking agent. When Tween 20 was reintroduced, the bands reappeared. These results demonstrate that, although Tween 20 is effective in blocking unbound sites on nitrocellulose membranes, this and other nonionic detergents may cause nonspecific binding to immunoblots of oak-pollen proteins. Thus, care must be used in the interpretation of immunoblots to detect allergens when Tween 20 is used as the blocking agent.

Animals↗
PubMed1987

Effect of serotonin (5-HT) and other monoamines on murine macrophages: modulation of interferon-gamma induced phagocytosis.

We have previously shown that serotonin (5-HT) suppresses interferon-gamma (IFN-gamma)-induced Ia expression. In the present report, we show that 5-HT as well as other monoamines, histamine and dopamine, modulate IFN-gamma-induced phagocytosis in murine bone marrow macrophages. The effect of 5-HT on IFN-gamma-induced phagocytosis varied according to the concentration of IFN-gamma to which the macrophages were exposed. At low concentrations of IFN-gamma, 5-HT augmented phagocytosis, whereas at high concentrations of IFN-gamma, 5-HT suppressed phagocytosis. At both low and high IFN-gamma concentrations the response to 5-HT was dose-related and occurred at physiologic concentrations; the half-maximal effect was 6 X 10(-7) M and 3 X 10(-7) M for low and high IFN-gamma concentrations, respectively. Both histamine and dopamine also augmented IFN-gamma (1 U/ml) induced phagocytosis, at half-maximal augmenting concentrations of 7 X 10(-8) M and 4 X 10(-7) M, respectively. The 5-HT effects were blocked by the 5-HT antagonists spiperone, ketanserin, LY53857, mCPP, and PAPP, but not by the histamine antagonists pyrilamine, chlorpheniramine, or cimetidine. Histamine augmentation of IFN-gamma-induced phagocytosis was blocked by the H1 antagonists pyrilamine and chlorpheniramine, but not by the H2 antagonist cimetidine. The dopamine effect was blocked by spiperone and pyrilamine, both of which have been shown to block dopaminergic effects in other systems. This data provides functional evidence that at least part of the modulation of IFN-gamma-induced phagocytosis by 5-HT occurs through a 5-HT receptor-mediated mechanism, and 5-HT, dopamine, and histamine modulate IFN-gamma-induced phagocytosis independently through their respective receptors.

Animals↗
PubMed1987

Stimulation of the phosphorylation of a 66,000-Da soluble tyrosine phosphoprotein by agents that activate or augment activation of human peripheral blood T lymphocytes.

Previous work from our laboratory has demonstrated the rapid phosphorylation on tyrosine of a 66-kDa soluble protein (TPP 66), when human T lymphocytes were incubated with the mitogenic lectins phytohemagglutinin or concanavalin A. To further explore the role of TPP 66 in lymphocyte activation we have utilized a variety of agents which either activate directly or augment the activation of the T cell and assayed their effect on TPP 66 phosphorylation. Wheat germ agglutinin, the divalent cation ionophore A23187, and phorbol myristate acetate have been demonstrated to activate human T cells and each of these agents also induced the rapid phosphorylation of TPP 66. In addition, epidermal growth factor and platelet-derived growth factor also induced the phosphorylation of TPP 66. Phosphorylation was seen at concentrations that are within the active dose range for all agents tested. Phosphorylation was on tyrosine, since base hydrolysis of TPP 66 followed by single-dimension high-voltage electrophoresis revealed radioactivity only in the area corresponding to authentic phosphotyrosine. These data demonstrate that a variety of agents which either directly activate human T lymphocytes or augment activation are associated with the tyrosine phosphorylation of TPP 66 and provide further evidence for a critical role for this protein in the control of lymphocyte activation.

Calcimycin↗
PubMed1987

Anaphylaxis to intravenous furosemide.

A patient with long-standing hypertension developed urticaria, angioedema, and hypotension within 5 minutes after the intravenous administration of furosemide. Immediate hypersensitivity was documented by positive skin tests to furosemide as well as to related sulfonamide-based drugs. This is the first finding of an anaphylactic reaction to furosemide and underscores the need to consider such adverse reactions when patients who are sensitive to other sulfonamide-containing drugs are being treated.

Adult↗
PubMed1987

Hypersensitivity reaction to desipramine.

Adverse reactions to the tricyclic antidepressant drugs imipramine and desipramine have been described and include eosinophilia, pulmonary infiltrates with eosinophilia, and elevated total serum IgE levels. The immunologic mechanism accounting for these adverse reactions has not been elucidated. This article describes a patient manifesting bronchospasm, profound eosinophilia, and elevated serum IgE levels after therapy with desipramine that resolved rapidly after withdrawal of the drug. Immunologic investigations failed to demonstrate specific IgE directed against a protein conjugate of desipramine but demonstrated the ability of desipramine to induce mast cell degranulation with direct intradermal skin challenges.

Desipramine↗
PubMed1987

Allergic reactivity of Parthenium hysterophorus (Santa Maria feverfew) pollen: an unrecognized allergen.

Parthenium hysterophorus (PH), a member of the Asteraceae, is a ubiquitous weed found in great profusion in the United States Gulf coast area. A plant survey carried out in Texas Gulf coast demonstrated that around Corpus Christi, PH is the most prevalent weed. We have studied known ragweed-sensitive patients living in Austin and Corpus Christi, by radioallergosorbent test (RAST) analysis for PH and Ambrosia psilostachya (AP; western ragweed). Nine of ten patients from Austin were RAST-positive to PH and AP; 8 of the 10 patients had positive scores to both pollen extracts. All of the 8 patients from Corpus Christi were positive to both PH and AP. The Austin group showed a significant correlation between AP and PH RAST scores while no correlation between RAST values was seen in the Corpus Christi group. RAST inhibition analysis in this highly selected population did reveal patients that had true sensitivity to PH pollen alone or who were sensitive to both PH and AP pollen. This suggests that in areas where it is highly prevalent PH pollen is an overlooked cause of allergic disease.

Allergens↗
PubMed1987

Allergic reactions to drugs.

Allergic reactions can result from virtually any drug. In the pharmacopeia the possibility of an allergic reaction to a drug must be considered in any instance when a patient reports an adverse effect to a drug. Although most allergic reactions to drugs are mild, they may be severe and at times fatal. Thus, it is imperative that every physician understand the spectra of drug allergy, the methods for diagnosing, and the means of treating allergic reactions to drugs. This review places drug allergies within the context of the over-all spectra of adverse reaction to drugs, defines the spectra of allergic reactions in terms of the organ systems involved and the severity of the reactions, suggests mechanisms for diagnosing drug allergy by historical evaluation, and where possible, by skin testing procedures and provocative dose challenges, and provides a variety of methods for reintroduction of drugs to which patients have been demonstrated to be allergic. The most important aspect of allergic reactions to drugs, however, is the understanding on the part of the physician that any drug has the potential for causing an allergic reaction and that extreme vigilance in terms of the diagnosis and treatment of these reactions is necessary.

Desensitization, Immunologic↗
PubMed1986

Induction of the tyrosine phosphorylation of a 66 kd soluble protein by DMSO in human peripheral blood T lymphocytes.

Previous studies from our laboratory have demonstrated that a 66 KD cytoplasmic protein (TPP 66) is newly phosphorylated on tyrosine when human peripheral blood T lymphocytes are incubated with a variety of agents that activate these cells or augment activation by known mitogens. Since DMSO has been shown to activate tyrosine specific protein kinases we have examined the role of this agent on the phosphorylation of TPP 66. 5 to 40% DMSO induced the phosphorylation of TPP 66 with the maximal increase in phosphorylation seen at 20%. Concentrations greater than 40% were inhibitory. Phosphorylation of TPP 66 in DMSO treated cells could be detected as early as 2 min following the addition of DMSO, with increased [32P]O4 incorporation over the next 60 min. The phosphorylation was on tyrosine residues demonstrated by base hydrolysis of TPP 66 extracted from the gels followed by single dimension high voltage electrophoresis. Since DMSO augments activation of T lymphocytes by lectins, this data provides further support for a critical role for the tyrosine phosphorylation of TPP 66 in the mediation or modulation of T lymphocyte activation.

Autoradiography↗
PubMed1986

Tyrosine phosphorylation of a 66,000 Mr soluble protein in lectin-activated human peripheral blood T lymphocytes.

Protein phosphorylation was studied in human T lymphocytes stimulated with the mitogenic lectins phytohemagglutinin (PHA) and concanavalin A (Con A). The T lymphocytes were prepared from the venous blood of normal volunteers, their intracellular ATP pools were labeled with [32P]orthophosphate, and protein phosphorylation was assayed in the soluble fraction by two-dimensional gel electrophoresis and autoradiography. When lymphocytes stimulated with PHA or Con A were compared to unstimulated control cells, there was a general increase in protein phosphorylation and the specific phosphorylation of a soluble protein with Mr = 64.9 to 69 KD and pI = 5.6 to 5.8. Phosphorylation of this protein, designated TPP-66, was observed as early as 2 min after the addition of lectin with a gradual increase in the level of phosphorylation over the next 120 min. In the majority of experiments, there was no phosphorylation seen in the unstimulated lymphocytes; however, in some experiments, there was appreciable phosphorylation, which was seen beginning 60 min after the labeling period. When the TPP-66 spot from stimulated lymphocytes was excised from gels, was eluted, and was subjected to limited base hydrolysis followed by single-dimension high voltage electrophoresis, the major phosphorylated residue migrated with phosphotyrosine. In some experiments, there was phosphorylation of serine residues in both the stimulated and control cells; tyrosine phosphorylation was never seen in the unstimulated cell population. These data suggest that, like other stimuli for cell growth, the induction of lymphocyte growth by lectins is associated with the activation of a tyrosine-specific kinase. Thus, tyrosine phosphorylation may play a key role in the transmission of the signal for lymphocyte growth from the exterior to the interior of the cell.

Autoradiography↗
PubMed1986

Anti-IgA antibody associated reactions to intravenous gammaglobulin in a patient who tolerated intramuscular gammaglobulin.

A patient with common variable immunodeficiency syndrome tolerated intramuscular IgG (which contains IgA) and an initial infusion with intravenous (IV) IgG, but developed reactions to subsequent IV IgG. High-titre, class-specific anti-IgA antibodies were detected suggesting immunization by the IgA-contaminated IV immunoglobulin. Subsequent IgG replacement was achieved with IgA-deficient plasma infusions. Patients who tolerate intramuscular IgG may not tolerate the IV preparations.

Antibodies, Anti-Idiotypic↗
PubMed1986

Inhibition of cell-mediated cytotoxicity by 2-cyclohexene-1-one: evidence for a role for glutathione and/or glutathione-protein interactions in cytolysis.

In order to explore the role of glutathione in cell-mediated cytotoxicity, we have examined the effect of the sulphydryl-reactive and glutathione-depleting agent 2-cyclohexene-1-one on antibody-dependent cellular cytotoxicity, spontaneous cell-mediated cytotoxicity, and cell-mediated lympholysis by human peripheral blood mononuclear cells. 2-Cyclohexene-1-one significantly inhibited (P less than 0.001) both antibody-dependent and spontaneous cell-mediated cytotoxicity using three different cell-line targets, at three different killer:target cell ratios (10:1, 25:1 and 50:1). Using K-562 cell-line targets, spontaneous cell-mediated cytotoxicity was inhibited by 2-cyclohexene-1-one with an ID50 of 0.71 X 10(-4) M-1.48 X 10(-4) M, while antibody-dependent cellular cytotoxicity was less sensitive to inhibition, and required slightly higher concentrations of 1.48 X 10(-4) M-3.98 X 10(-4) M to achieve 50% inhibition. Similar results were seen with human colon tumour cell-line and Chang liver cell-line cells as targets. Maximal inhibition occurred when 2-cyclohexene-1-one was added to the cytotoxicity assay 60 min prior to, at the start of, or within the first 60 min of a 4-hr assay; inhibition of cytotoxicity occurred with pretreatment of effector cells; and no inhibition of cytotoxicity was observed with pretreatment of target cells. Both the allogeneic mixed leucocyte reaction and cell-mediated lympholysis were also significantly inhibited (P less than 0.001) by 2-cyclohexene-1-one. These studies demonstrate that 2-cyclohexene-1-one is an effective inhibitor of cell-mediated cytotoxicity and suggest that glutathione, specific glutathione-protein interactions, or protein-bound sulphydryl groups are involved in allowing cells to carry out cytolysis.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1985

Inhibition of lectin-induced lymphocyte activation by 2-cyclohexene-1-one: analysis of DNA synthesis in individual cells by BUdR quenching of Hoechst 33258.

A novel technique utilizing the quenching of fluorescence Hoechst 32258 by bromodeoxyuridine (BUdR) was used to investigate the effect of depressed glutathione (GSH) on the activation of human peripheral blood lymphocytes by phytohemagglutinin (PHA) or concanavalin A (con A). This technique allows the quantification of DNA synthesis in individual cells. Lymphocytes were purified by Ficoll-Hypaque density gradient centrifugation and treated with 5 X 10(-5) M to 1 X 10(-6) M 2-cyclohexene-1-one (2-CHX-1), a reagent which specifically depletes intracellular GSH, and/or interferes with GSH-protein interactions, and 25 micrograms/ml BUdR in the presence or absence of PHA or con A. At 72 h lymphocyte smears were stained with Hoechst 33258 and examined using a computer controlled microscope photometer. When DNA synthesis was assayed using BUdR quenching two populations of lymphocytes were noted; a population which incorporated little or no BUdR (unactivated) and a population which incorporated BUdR sufficient to quench 33258 fluorescence by approximately 35%. Cells treated with graded doses of 2-CHX-1 which reduced glutathione levels by 10-90%, showed a progressive loss of cells from the activated population and the appearance of these cells in the inactivated population. Statistical analysis of the frequency histograms demonstrated that there were no cells which incorporated an intermediate amount of BUdR. This data demonstrates that depressed intracellular GSH or inhibition of GSH-protein interactions inhibits an early step in the biochemical sequence of events leading to DNA synthesis but does not inhibit the DNA synthetic process per se.

Benzimidazoles↗