In Vitro Antiid Manipulation Of Autoantibodies

In vitro studies demonstrating modulation by anti-Id support the theory that anti-Id may modulate autoantibody activity in vivo.

Kim et al. [20] have demonstrated that anti-DNA production by anti-DNA-secreting hybridomas can be inhibited by the addition of anti-idiotypes to anti-DNA. In this study, a series of anti-DNA antibody producing hybridomas were obtained by fusing spleen cells from 6-month old MRL/lpr autoimmune prone mice with P3X63-Ag8 myeloma cells. Rabbit anti-idiotype antibodies specific for several of the hybridoma proteins were prepared. It was shown that the anti-Id antibody inhibited immunoglobulin secretion by the hybridoma cells in an Id-specific manner. Inhibition of antibody production was not due to a cytotoxic effect, since the anti-Id, in fact, stimulated proliferation of the hybridoma cells.

In order to assess the anti-Id network in murine experimental autoimmune encephalomyelitis (EAE), Id-bearing monoclonal antibodies (mAb) to human myelin basic protein (MBP) peptide acetyl 1-9, as well as mAb anti-Id, were developed in EAE-susceptible PL/j mice (H-2u) [21]. These mice recognize MBP residues acetyl 1-9 as an encephalitogenic determinant. Reactivities of PL/j Id-bearing mAbs to MBP and to MBP peptides are identical to those of mAbs generated against the same MBP peptide in EAE-resistant BALB/C mice (H-2d), even though isotypes of the mAbs differed. By using an inhibitory ELISA and immunoblotting, it was demonstrated that one PL/j mAb anti-id recognized a public or framework Id, whereas another PL/j mAb-anti Id was directed to a private Id more restricted to the paratopic site. Two Id-bearing PL/j mAbs shared a cross-reactive Id (IdX) on the light chain, and an interstrain IdX was present on both the heavy and light chains of mAbs raised in PL/j and BALB/C mice to the same MBP peptide. The PL/j mAb anti-Id was capable of cross-regulating the production of Id-bearing mAbs by hybridomas across murine strains. These findings suggest that manipulation of the Id network may provide a means for modifying autoimmune demyelinating diseases of the central nervous system [21].

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