Analysis of the repertoire of anti-NP antibodies in C57BL/6 mice by cell fusion. I. Characterization of antibody families in the primary and hyperimmune response

Spleen cells from C57BL/6 mice sensitized to the hapten (4-hydroxy-3-nitrophenyl)acetyl (NP) were hybridized with myeloma cells, and a variety of hybrid cell lines was isolated which secreted homogeneous anti-NP antibodies. The antibodies were purified by affinity chromatography and their chain composition, affinity and fine specificity were determined. All antibodies recovered from the primary immune response carried λ light and μ or γ1 heavy chains. Their variable portions were nonidentical but similar in terms of hapten-binding specificity with a higher affinity for the cross-reacting haptens (4-hydroxy-3,5-dinitro-phenyl)acetyl (NNP) and (4-hydroxy-5-iodo-3-nitro-phenyl)acetyl (NIP) than for the homologous hapten NP. This heteroclitic property as well as the presence of λ, μ and γ1 chains are characteristic for primary anti-NP sera of C57BL/6 mice. In contrast, four families of anti-NP antibodies, each with a characteristic fine specificity pattern, were found among the hybrid cell antibodies derived from the hyperimmune anti-NP response. The antibodies of one of these families were related to the antibodies recovered from the primary immune response in that they were heteroclitic and carried λ light chains. All members of the other groups expressed k chains and were nonheteroclitic. The finding of well-defined antibody families in this system and the isolation of their members enable us to approach the problem of V gene expression and diversification in a new way.