Characterization of the NPa idiotype through the analysis of monoclonal BALB/c anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) antibodies

Abstract

Spleen cells from BALB/c mice were fused with the nonsecreting myeloma line X63.Ag8.6.5.3 seven days after immunization with NP-CG (4-hydroxy-3-nitrophenyl) acetyl-chicken gamma-globulin. The hybrid cell lines obtained were analyzed for heavy and light chain distribution, fine specificity, and idiotype. The majority of monoclonal antibodies possessed either gamma 1 or mu chains. The distribution of L chains among these antibodies was approximately half lambda and half kappa . Thirteen monoclonal antibodies were grown as ascites tumors in mice. Examination of their fine specificity patterns showed that all of the lambda antibodies are heteroclitic and have similar fine specificity patterns. Five of the seven kappa antibodies are also heteroclitic, but their fine specificity patterns are more heterogeneous than those of the lambda antibodies. Polyspecific anti-idiotypic sera directed against pooled primary serum antibody (R a-NPa) or against individual monoclonals were used for idiotypic characterization of the monoclonal antibodies. The Ra-NP bound all of the lambda antibodies but none of the kappa antibodies suggesting that the kappa antibodies may be much more heterogeneous and were therefore not recognized in the presence of the more homogeneous lambda antibodies. Further idiotypic analysis demonstrated that the lambda antibodies, although no two are identical, are a very homogeneous group of antibodies which cross-react with one another but not with the kappa antibodies. Some, but not all, of the kappa antibodies cross-react with each other although none are cross-reactive with the lambda antibodies. Because the lambda-associated idiotype is recognized by the R a-NPa and its characteristics are similar to that of the C57BL/6 major idiotype (NPb), it is referred to as NPa. There may be a second major idiotype associated with at least some of the kappa antibodies.