Immune hemolytic assay for identification of human anti-dsDNA antibodies with DNA-coated red blood cells as target cells. 1988

M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
Laboratory of Medical Immunology, State University Hospital (Rigshospitalet), Copenhagen, Denmark.

A hemolytic assay was developed with the primary aim of being able to identify human lymphocytes producing anti-dsDNA antibodies found in patients with systemic lupus erythematosus (SLE). The coating of sheep red-blood cells with DNA was performed after precoating the cells with poly-L-lysine. The DNA-SRBC were lysed by anti-DNA antibodies from SLE sera, and the percent hemolysis was found to correlate with the anti-DNA activity demonstrated by the Farr assay (r = 0.87). Single-stranded DNA at the surface of the coated cells could be removed after digestion with nuclease S1. The effect of the digestion was verified by SLE serum specific for single-stranded DNA. With slight modifications, the target cells may be used to determine not only the titer of anti-DNA antibodies but also the complement-consumption and immunoglobulin classes of the anti-DNA antibodies.

UI MeSH Term Description Entries
D008180 Lupus Erythematosus, Systemic A chronic, relapsing, inflammatory, and often febrile multisystemic disorder of connective tissue, characterized principally by involvement of the skin, joints, kidneys, and serosal membranes. It is of unknown etiology, but is thought to represent a failure of the regulatory mechanisms of the autoimmune system. The disease is marked by a wide range of system dysfunctions, an elevated erythrocyte sedimentation rate, and the formation of LE cells in the blood or bone marrow. Libman-Sacks Disease,Lupus Erythematosus Disseminatus,Systemic Lupus Erythematosus,Disease, Libman-Sacks,Libman Sacks Disease
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004720 Endonucleases Enzymes that catalyze the hydrolysis of the internal bonds and thereby the formation of polynucleotides or oligonucleotides from ribo- or deoxyribonucleotide chains. EC 3.1.-. Endonuclease
D004912 Erythrocytes Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN. Blood Cells, Red,Blood Corpuscles, Red,Red Blood Cells,Red Blood Corpuscles,Blood Cell, Red,Blood Corpuscle, Red,Erythrocyte,Red Blood Cell,Red Blood Corpuscle
D006461 Hemolysis The destruction of ERYTHROCYTES by many different causal agents such as antibodies, bacteria, chemicals, temperature, and changes in tonicity. Haemolysis,Extravascular Hemolysis,Intravascular Hemolysis,Extravascular Hemolyses,Haemolyses,Hemolyses, Extravascular,Hemolyses, Intravascular,Hemolysis, Extravascular,Hemolysis, Intravascular,Intravascular Hemolyses
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000937 Antigen-Antibody Reactions The processes triggered by interactions of ANTIBODIES with their ANTIGENS. Antigen Antibody Reactions,Antigen-Antibody Reaction,Reaction, Antigen-Antibody,Reactions, Antigen-Antibody
D001323 Autoantibodies Antibodies that react with self-antigens (AUTOANTIGENS) of the organism that produced them. Autoantibody
D015719 Single-Strand Specific DNA and RNA Endonucleases Enzymes that catalyze the endonucleolytic cleavage of single-stranded regions of DNA or RNA molecules while leaving the double-stranded regions intact. They are particularly useful in the laboratory for producing "blunt-ended" DNA molecules from DNA with single-stranded ends and for sensitive GENETIC TECHNIQUES such as NUCLEASE PROTECTION ASSAYS that involve the detection of single-stranded DNA and RNA. Single Strand Specific DNA and RNA Endonucleases
D066298 In Vitro Techniques Methods to study reactions or processes taking place in an artificial environment outside the living organism. In Vitro Test,In Vitro Testing,In Vitro Tests,In Vitro as Topic,In Vitro,In Vitro Technique,In Vitro Testings,Technique, In Vitro,Techniques, In Vitro,Test, In Vitro,Testing, In Vitro,Testings, In Vitro,Tests, In Vitro,Vitro Testing, In

Related Publications

M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
November 1999, Nihon rinsho. Japanese journal of clinical medicine,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
April 2013, Transfusion,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
January 1985, Scandinavian journal of rheumatology,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
June 1986, Proceedings of the National Academy of Sciences of the United States of America,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
January 1981, Transfusion,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
January 1981, Tissue antigens,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
January 2009, Xenotransplantation,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
November 1973, Cellular immunology,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
July 2012, The Journal of dermatology,
M K Loftager, and C Koch, and P Hellung-Larsen, and V Andersen
January 1965, Vox sanguinis,
Copied contents to your clipboard!