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1.
Appl Biochem Biotechnol ; 23(1): 1-13, 1990 Jan.
Article in English | MEDLINE | ID: mdl-2301946

ABSTRACT

In order to investigate relationships between human carcinogenesis and dietary carcinogens, one hybridoma cell line secreting a monoclonal antibody against 2-amino-3-methylimidazo(4,5-f)quinoline (IQ), a dietary carcinogen, was produced by fusing splenocytes from Balb/c mice immunized with IQ-Lysine(Lys)-Ascaris protein conjugate. The subclass of monoclonal anti-IQ antibody was determined by double immunodiffusion using culture medium and identified as IgG1. Monoclonal anti-IQ antibody was purified from ascites fluids of Balb/c mice with affinity chromatography on Protein A-Sepharose CL4B and analyzed concerning its cross-reactivity and sensitivity with RIA. Finally, we showed that our monoclonal antibody recognized IQ, 2-amino-3,4-dimethylimidazo(4,5-f)quinoline (MeIQ) and several beta-carbolines more intensely and that the sensitivity to IQ was 23 nmol in 50% displacement.


Subject(s)
Antibodies, Monoclonal , Carcinogens , Diet , Quinolines/immunology , Animals , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/isolation & purification , Antibody Specificity , Antigens/immunology , Ascitic Fluid/immunology , Carbolines/immunology , Chromatography, Affinity , Hybridomas/immunology , Immunoglobulin G/immunology , Mice , Mice, Inbred BALB C , Molecular Weight , Radioimmunoassay
2.
Histochemistry ; 80(6): 597-601, 1984.
Article in English | MEDLINE | ID: mdl-6469716

ABSTRACT

Recently (Pecci Saavedra et al. 1982; Brusco et al. 1982, 1983) we have showed that the actual specificity of the rabbit anti-5-HT antibodies, is for the beta-carboline derivatives of 5-HT as a result of cyclization of the lateral chain. We explained this as resulting from the use of formaldehyde which acted both as a fixative in the preparation of the tissues, and as the coupling agent in the preparation of the immunogen. Following this line we have fixed several brain stem specimens with 0.5% p-benzoquinone; 3% glutaraldehyde; 4% paraformaldehyde plus 0.25% glutaraldehyde and compare the results with tissues fixed in 4% paraformaldehyde. Glutaraldehyde and p-benzoquinone do not produce cyclization of 5-HT but immobilize monoamines in situ. As expected, the antibodies applied according to the PAP technique did not stain the neuronal bodies of the raphe system, known to contain 5-HT when 3-4% glutaraldehyde or 0.5% p-benzoquinone were used. Good staining was obtained with 4% paraformaldehyde alone or with 4% paraformaldehyde plus 0.25% glutaraldehyde. A quantitative assay of the spot test of Larsson (1981) was devised for measuring in vitro the inhibitory effects of 5-HT, of the 5-HT-BSA complex and of the cyclic derivative, 6-OH-1,2,3,4-tetrahydro-beta-carboline. The results confirmed that the avidity of the antiserum is much greater for the cyclic derivatives contained in the 5-HT-BSA complex and for 6-OH-1,2,3,4-tetrahydro-beta-carboline than for 5-HT. It is concluded that the formation of a new ring by the lateral chain of 5-HT is responsible of the in-vitro and in the tissue immunoreactivity of the anti-5-HT-antibodies.


Subject(s)
Antibody Specificity , Central Nervous System/metabolism , Serotonin/immunology , Animals , Carbolines/immunology , Fixatives , Formaldehyde , Histocytochemistry , Immunochemistry , Rabbits , Rats , Serotonin/metabolism
3.
Farmakol Toksikol ; 43(1): 62-5, 1980.
Article in Russian | MEDLINE | ID: mdl-6103820

ABSTRACT

The effect of antihistaminic drugs, histamine and their combinations on the development of anaphylactic shock and the period of repeated resolution doses was studied in rabbit experiments. It was established that administration of dimedrol during sensibilization aggravated while diazolin and histamine attenuated the course of anaphylactic shock and periods of repeated resolution injections. Simultaneous administration of antihistaminic agents and histamine enhanced the allergization of the body. Under conditions reported dimedrol prevented hypertrophy of the adrenals that occurred in response to sensibilization. Meanwhile diazolin did not interfere with the reaction development and histamine stimulated it. Combined administration of the antihistaminic agents and histamine provoked an inhibition of the above reaction of the adrenals to body sensibilization.


Subject(s)
Drug Hypersensitivity/etiology , Histamine H1 Antagonists/immunology , Histamine/immunology , Anaphylaxis/chemically induced , Animals , Carbolines/immunology , Diphenhydramine/immunology , Dose-Response Relationship, Immunologic , Immune Sera/administration & dosage , Immunization , Naphthalenesulfonates/immunology , Rabbits , Recurrence , Swine/immunology
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