ABSTRACT
Binding of ssDNA and dsDNA to thioglycollate-stimulated peritoneal macrophages derived from normal CBA mice was increased not only by ssDNA-specific antibody but also by albumin and gamma-globulin preparations. Fresh mouse serum inhibited ssDNA binding, even if the ssDNA and serum were preincubated at 37 degrees C. Heat decomplemented mouse serum also depressed DNA binding. These results are significant to interpretations of DNA clearance rate data from serum or blood, and subsequent inferences concerning blood DNA levels, autoimmune response, and systemic lupus erythematosus.
Subject(s)
Blood , DNA/metabolism , Macrophages/metabolism , Thioglycolates/pharmacology , Animals , DNA, Single-Stranded/metabolism , Macrophages/drug effects , Mice , Mice, Inbred CBA , Peritoneum/cytology , Serum Albumin/pharmacology , Serum Globulins/pharmacologyABSTRACT
A common requirement for chemical-induced allergy, carcinogenesis and certain forms of liver injury appears to be the ability of the chemical to react with protein and/or nucleic acid to form covalent conjugates. Tests to detect immune responses to such conjugates can provide useful information concerning the propensity of an individual to convert normally innocuous chemicals to immuno-reactive forms under physiological conditions and may provide an indicator of the risk for developing one of the above disorders. They will enable offending substances and their active metabolites to be identified, the status of diseases which they induce determined, and the degree to which exposure has been avoided monitored. Very few studies have been published relating to this important problem. Implications of this approach are discussed.
Subject(s)
Drug Hypersensitivity/immunology , Immunity/drug effects , Neoplasms/chemically induced , Animals , Antibody Formation/drug effects , Biotransformation , Humans , In Vitro Techniques , Liver/drug effects , Microsomes, Liver/metabolism , Molecular Weight , Nucleic Acids/metabolism , Protein Binding , Rats , Rats, Inbred Strains , Tartrazine/toxicityABSTRACT
The binding of DNA-antibody complexes to thioglycollate-induced mouse peritoneal macrophages is inhibited by fresh but not by decomplemented normal mouse serum. Binding to macrophage complement receptors was not observed.
Subject(s)
Antibodies/metabolism , DNA, Single-Stranded/metabolism , Macrophages/metabolism , Animals , DNA, Single-Stranded/immunology , Female , Macrophages/ultrastructure , Male , Mice , Mice, Inbred CBA , Peritoneal Cavity/cytology , Receptors, Fc/metabolism , TemperatureSubject(s)
Azo Compounds/adverse effects , Drug Hypersensitivity/immunology , Immunoglobulin D/biosynthesis , Immunoglobulin E/biosynthesis , Tartrazine/adverse effects , Animals , Binding Sites, Antibody , Drug Hypersensitivity/etiology , Ethanolamines/immunology , Humans , Rabbits , Radioimmunoassay/methods , Serum Albumin/immunology , Sulfanilic Acids/immunology , Tartrazine/immunologyABSTRACT
The clinical diagnosis of hypersensitivity to the food dye tartrazine in 16 subjects correlated with levels of tartrazine-specific IgD antibodies determined by a solid phase radioimmunoassay inhibition test. This is the first study associating hypersensitivity to a low molecular weight chemical with an antibody response of the IgD class. There was little or no correlation between clinical sensitivity and IgE antibodies.
Subject(s)
Antibody Specificity , Azo Compounds/adverse effects , Drug Hypersensitivity/immunology , Immunoglobulin D , Tartrazine/adverse effects , Antibody Formation , HumansABSTRACT
Soluble Epstein--Barr virus (EBV) associated antigens of P3HR-1 and Raji cell lines, which fix complement in the presence of human sera containing antibody against EBV--VCA antigens, were partially purified by fractionating centrifuged cell lysates by the method of immunoadsorbents or by precipitation with ammonium sulphate or glycine--HCl. Purification conditions were limited by the instability of both antigen and antibody activity. The antibody was inactivated under all the conditions commonly used to dissociate antigen--antibody complexes. The antigen was inactivated at pH 2-3, and was irreversibly bound to Sephadex G-200 conjugates which had not been exposed to large excesses of serum.
Subject(s)
Antigens, Viral/isolation & purification , Herpesvirus 4, Human/immunology , Adsorption , Antibodies, Viral , Antigen-Antibody Complex , Cell Line , Chemical Precipitation , Complement Fixation Tests , Electrophoresis, Polyacrylamide Gel , Humans , Hydrogen-Ion ConcentrationABSTRACT
The soluble complement-fixing antigen of the P3HR-1 Burkitt's lymphoma cell line, identified with human serum containing antibody against Epstein-Barr virus viral capsid antigen, loses activity under a variety of conditions. The major characteristic reported here is retention of activity on exposure to specific physical or chemical conditions followed by loss of activity after subsequent neutral dialysis. Antigen of untreated cell lysates, which retains activity after dialysis against large volumes of neutral buffers or 0.14 M NaCl, will lose activity if the lysate is first heated to 56 degrees or if the lysate is exposed to acid perchlorate, and the resulting precipitate and supernatant are redialyzed separately to neutral pH. The P3HR-1 soluble complement-fixing antigen appears to be an aggregate including a labile, dissociable component.
Subject(s)
Antigens, Neoplasm/analysis , Burkitt Lymphoma/immunology , Antibodies, Viral , Buffers , Cell Line , Chemical Phenomena , Chemistry , Complement Fixation Tests , Dialysis , Herpesvirus 4, Human/immunology , Hot Temperature , Humans , Hydrochloric Acid , Hydrogen-Ion Concentration , PerchloratesABSTRACT
Soluble complement-fixing (CF) antigens of the virus-producing and virus-nonproducing P3HR-1 and RAJI Burkitt lymphoma cell lines and Hk-Ly-28 nasopharyngeal cancer cell line, identified by use of viral capsid antigen-positive human sera, could readily be distinguished by differences in their stability to chemical and physical conditions. The P3HR-1 soluble CF antigen lost titer when heated or exposed to acid perchlorate under conditions in which RAJI and Hk-Ly-28 soluble antigens were stable. Differences in solubility were also found. These results contribute not only to the interpretation of the relationship of Epstein-Barr virus (EBV)-associated components of RAJI to P3HR-1 and Hk-Ly-28 cell lysates but also to the selection of conditions for the development of identification tests and purification procedures for CF antigens and antibodies of Burkitt's lymphoma, nasopharyngeal cancer, and other EBV-associated tumors.
