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[Advances in immunoserology tests in clinical medicine: autoantibodies, immune complexes and DNA analysis]
1Ohta General Hospital Foundation, Institute of Rheumatic Diseases, Koriyama 963-8004.
I reviewed first the history of detection for autoantibodies and the methods to detect the circulating immune complexes. Then I presented several unusual tests for detecting autoantibodies which I had experienced. They are agglutination tests in agarose gel using the particulate antigens of human tissues or lipid antigens and the mixed agglutination test using the cultured cells. The characteristics of rheumatoid factor(RF) were analyzed by the following methods: 1) solid-phase radioimmunoassay for IgG-RF using the formalinized sheep RBC, 2) mixed agglutination test for IgG-RF on the slide glass smeared with the sensitized sheep RBC, 3) hemolysis in agarose gel for competitive reaction of RF with complement to the IgG hemolysin and 4) ELISA for detecting the complement-fixation of the monoclonal IgM-RF. The IgG-Fc and C3b receptors on the tissue sections of the mammalian aorta were detected by an adhesion of the IgG-sensitized or C3b-bound RBC. DNA-analysis studies using the molecular biology techniques which were done in the Department of Internal Medicine II, Fukushima Medical University were presented. 1) Nucleic acid sequences of the cloned DNA polypeptide fragments in the serum of patients with systemic lupus erythematosus. 2) SSCP analysis for clonality of Vb repertoires in T cell receptors in the patients with rheumatoid arthritis, primary biliary cirrhosis or CREST syndrome. 3) Detection of mRNA of TNFa and Fas ligand in the mononuclear cells(FICL-PCR) in the synovial fluids of RA patients.
I reviewed first the history of detection for autoantibodies and the methods to detect the circulating immune complexes. Then I presented several unusual tests for detecting autoantibodies which I had experienced. They are agglutination tests in agarose gel using the particulate antigens of human tissues or lipid antigens and the mixed agglutination test using the cultured cells. The characteristics of rheumatoid factor(RF) were analyzed by the following methods: 1) solid-phase radioimmunoassay for IgG-RF using the formalinized sheep RBC, 2) mixed agglutination test for IgG-RF on the slide glass smeared with the sensitized sheep RBC, 3) hemolysis in agarose gel for competitive reaction of RF with complement to the IgG hemolysin and 4) ELISA for detecting the complement-fixation of the monoclonal IgM-RF. The IgG-Fc and C3b receptors on the tissue sections of the mammalian aorta were detected by an adhesion of the IgG-sensitized or C3b-bound RBC. DNA-analysis studies using the molecular biology techniques which were done in the Department of Internal Medicine II, Fukushima Medical University were presented. 1) Nucleic acid sequences of the cloned DNA polypeptide fragments in the serum of patients with systemic lupus erythematosus. 2) SSCP analysis for clonality of Vb repertoires in T cell receptors in the patients with rheumatoid arthritis, primary biliary cirrhosis or CREST syndrome. 3) Detection of mRNA of TNFa and Fas ligand in the mononuclear cells(FICL-PCR) in the synovial fluids of RA patients.