Immune hemolytic assay for identification of human anti-dsDNA antibodies with DNA-coated red blood cells as target cells.
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.
Loftager
M K
MK
Laboratory of Medical Immunology, State University Hospital (Rigshospitalet), Copenhagen, Denmark.
Koch
C
C
Hellung-Larsen
P
P
Andersen
V
V
D000937:Antigen-Antibody Reactions; D001323:Autoantibodies / Q000032:analysis*; D004247:DNA / Q000276:immunology*; D004720:Endonucleases / Q000494:pharmacology; D004912:Erythrocytes; D006461:Hemolysis; D006801:Humans; D066298:In Vitro Techniques; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D015719:Single-Strand Specific DNA and RNA Endonucleases
D001323:Autoantibodies; D004247:DNA; D004720:Endonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases