Production of Monoclonal Anti-idiotypic Antibody to Monoclonal Anti-DNA Antibody

Abstract

It has been thought that autoimmune diseases like systemic lupus erythematosus and

rhumatoid athritis are closely associated with anti-DNA antibodies (Abs). In studies of the control for anti-DNA Ab generation, an understanding of the regulatory mechanisms by anti-idiotypic Abs that influence the production of anti-DNA Abs would be facillitated by the availability of the hybridomas producing the pairs of DNA-specific and anti-DNA's idiotope-specific monoclonal antibodies (mAbs). We have produced a series of anti-DNA mAbs and then monoclonal anti-idiotypic Ab directed against idiotypic determinant of the 3D8 mAb that has the highest affinity to dsDNA and

SSDNA among the anti-DNA mABs that we had obtained. The spleen cells of the MRL-lpr/lpr, autoimmune prone, mice were fused with P3×63Ag8.653 myeloma cells to obtain anti-DNA Ab secreting hybridomas. Out of the fourteen clones that showed strong binding to ssDNA, four clones had cross-reactivity with dsDNA whereas none of these clones reacted with left-handed z-DNA. The binding activities of the anti-DNA mAbs to various synthetic polynucleotide sequences were different respectively.

Anti-idiotypic mAbs were generated by the fusion of myeloma cells and spleen cells

from the Balb/c mice immunized with 3D8-Fab. We have produced two anti-idiotypic

mAbs, B7 (IgG2a/ ) and 02F3 (IgM/ ), which were specific to 3D8-Fab and cloned the

variable region of the heavy chain from the O2F3 hybridoma.