title: Second messenger-induced modulation of Thy-1 antigen expression: an ELISA study using murine thymic and neuroblastoma cells.
abstract: Freshly isolated cells of murine thymuses and in vitro cultured murine neuroblastoma cells were seeded into microplates, treated with either phorbol myristate acetate (PMA) or dibutyryl-cyclic adenosine monophosphate (DBcAMP), and then processed for enzyme immunoassay to analyze with G4 monoclonal antibody the expression of Thy-1.2 antigen epitope. Pretreatment of thymic cells with PMA resulted in little, if any, decrease of Thy-1 expression, while treatment of these cells with DBcAMP caused a significant down-modulation of the epitope. DBcAMP did not affect binding of another murine IgG antibody to the thymic cells. Modulation of the epitope on thymic cells caused by DBcAMP was dose-dependent with maximal effect seen at the drug concentration of 10(-4) M. However, at various doses of DBcAMP (10(-6) to 10(-3) M) we were unable to detect any significant shift of Thy-1 expression on neuroblastoma cells. Though mechanisms of the above phenomena need further elucidation, we conclude that cellular ELISA may provide a useful alternative to more commonly used cytofluorimetric studies for the analysis of immune cell-surface antigen expression and its pharmacological and physiological modulation.
authors.affiliation: Department of Neurochemistry, A.A.Bogomoletz Institute of Physiology, Kiev, Ukraine, USSR.
authors.forename: G V
authors.initials: GV
authors.lastname: Pinchuk
authors.forename: L N
authors.initials: LN
authors.lastname: Pinchuk
authors.forename: A V
authors.initials: AV
authors.lastname: Grishchenko
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism*; D003994:Bucladesine / Q000494:pharmacology; D015536:Down-Regulation; D004797:Enzyme-Linked Immunosorbent Assay; D066298:In Vitro Techniques; D051379:Mice; D008808:Mice, Inbred CBA; D009447:Neuroblastoma / Q000378:metabolism; D015290:Second Messenger Systems / Q000276:immunology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018800:Thy-1 Antigens; D013950:Thymus Gland / Q000166:cytology / Q000378:metabolism
chemical_list: D000954:Antigens, Surface; D018800:Thy-1 Antigens; D003994:Bucladesine; D013755:Tetradecanoylphorbol Acetate