Subject(s)
Defibrillators, Implantable , Nerve Block/methods , Pain, Postoperative/prevention & control , Sternotomy/adverse effects , Tachycardia, Ventricular/therapy , Adolescent , Anesthetics, Local/administration & dosage , Humans , Lumbar Vertebrae , Male , Pain Measurement , Pain, Postoperative/diagnosis , Pain, Postoperative/etiology , Paraspinal Muscles/diagnostic imaging , Paraspinal Muscles/innervation , Treatment Outcome , Ultrasonography, InterventionalABSTRACT
BACKGROUND: The induction of nonanaphylactogenic 'blocking' IgG antibodies capable of inhibiting the IgE/allergen interaction represents a favorable therapeutic concept for type I allergy. However, IgG antibodies to allergens may block or enhance specific IgE binding, depending on the recognized epitope. Taking the major birch pollen allergen Bet v 1 as a model, we developed a strategy for the precise induction of IgG antibodies of a desired epitope specificity. METHODS: Random phage display peptide libraries were applied to define peptide structures mimicking natural epitopes (mimotopes) of Bet v 1. Selections were performed with BIP 1, a murine monoclonal antibody known to enhance the IgE binding to Bet v 1, and with anti-Bet v 1 IgE purified from patients' sera. The characterized Bet v 1 mimotopes were used to localize the corresponding epitope at the surface of Bet v 1 by a computer-aided mathematical approach based on the three-dimensional structure and the chemical character of the amino acids. The Bet v 1 mimotopes were further used to immunize BALB/c mice. The specificity of the induced antibodies was tested by immunoblotting and inhibition assays. RESULTS: With the three-dimensional epitope search it became possible to localize a discontinuous IgE epitope on the surface of Bet v 1 in a substantial distance from the IgG epitope of the monoclonal antibody BIP 1. Moreover, we could demonstrate that phage displaying mimotopes are immunogenic vectors for the precise induction of epitope-specific IgG. Immunization with BIP 1 mimotopes induced IgG enhancing the IgE binding to Bet v 1, whereas immunization with IgE mimotopes resulted in IgG capable of blocking human IgE binding in vitro. CONCLUSION: Allergen mimotopes can be used for the induction of anti allergen IgG of desired specificity. We propose that mimotope immunotherapy based on IgE mimotopes generated by biopannings may represent a future concept for therapy of type I allergy.
Subject(s)
Allergens , Epitopes/immunology , Immunoglobulin E/immunology , Immunoglobulin G/biosynthesis , Plant Proteins/immunology , Animals , Antibody Specificity , Antigens, Plant , Female , Mice , Mice, Inbred BALB C , Models, Immunological , Molecular Mimicry , Peptide Library , Pollen/immunologyABSTRACT
Peripheral blood mononuclear cells (PBMC) from 40 patients with gastrointestinal carcinoma (GIC), 13 patients with primary carcinoma in other localizations(non-GIC), and from 57 apparently healthy donors were isolated by Ficoll-Paque gradient centrifugation. The separated cells were stained with several monoclonal antibodies and subjected to analysis on a fluorescence-activated cell sorter. A decreased percentage of PBMC expressing T cell antigens was noted amongst GIC patients, and was mainly due to a reduction of the Leu 2a subset, thus, leading to an increase in the Leu 3a/Leu 2a ratio from 1.4 to 2.1 Non-GIC patients had decreased numbers of both T helper and suppressor cells. Amongst PBMC from GIC and non-GIC patients a statistically increased percentage of cells expressed LeuM 2 (P less than 0.001), LeuM 3 (P less than 0.001), OKM 1 (P less than 0.005), VEP 9 (P less than 0.001), and HLA-DR (P less than 0.001) antigens compared to healthy controls. The percentage of cells bearing these monocyte/macrophage antigens correlated well with the number of cells having monocyte morphology, stained for non-specific esterase, phagocytosed latex particles, and expressed Fc IgG receptor. Our results demonstrate clearly that tumor-bearing patients have an increased relative number of monocytes. The data suggest that cells of the macrophage lineage may be involved in defense mechanisms and changes of the immune system evoked by various tumors.
Subject(s)
Gastrointestinal Neoplasms/blood , Monocytes/cytology , Adolescent , Adult , Aged , Aged, 80 and over , Antigens, Surface/analysis , Flow Cytometry , Humans , Liver Neoplasms/blood , Middle Aged , Phenotype , Surface PropertiesABSTRACT
The cortical layer of the vitreous body of the eye contains a homogeneous population of hyalocytes. These cells were prepared from human autopsy eyes under sterile conditions by incubating the vitreous gel in tissue culture medium containing 300 micrograms/ml hyaluronidase. With the use of trypan blue staining, 60% to 90% of the cells were viable; they stained strongly for an intracellular nonspecific esterase with alpha-napthylacetate as a substrate. This staining could not be inhibited by sodium fluoride. The hyalocytes showed adherence to glass and plastic surfaces and phagocytosed latex spheres of 1.1 micrometer diameter. On their surface, receptors for IgG and complement components could be demonstrated with a rosette-forming technique using sensitized sheep erythrocytes. All these features strongly support the assumption that hyalocytes are mature cells of the mononuclear phagocyte system.
