The range and fine specificity of the anti-hapten immune response: phylogenetic studies

Heterodontus francisci (horned shark) and Pseudopleuronectes americanus (winter flounder) were immunized with furyl-oxazolone (furyl-Ox) and phenyl-oxazolone (phenyl-Ox) coupled either to bacteria or protein carriers. The antibodies produced were measured by inactivation of furyl- or phenyl-Ox conjugated bacteriophage, and their affinity and fine specificity were estimated by inhibition of phage inactivation with a series of structurally related hapten analogues. In both species, post-immunization peak titres were 100 to 2000 times higher than preimmunization titres. A number of unique features distinguished Heterodontus antibodies from Pseudopleuronectes or mammalian antibodies. Heterondontus antibodies exhibited a lower affinity for the immunizing hapten (furyl-Ox or phenyl-Ox) and a reduced ability to distinguish the homologous immunogenic hapten from its structural analogues. In addition, Heterodontus antibodies exhibited a lower level of inter-individual variation in affinity and fine specificity than did Pseudopleuronectes or mammalian IgM antibodies; this was especially prominent in anti-furyl-Ox responses. Typically the affinity and fine specificity of Heterodontus antibodies did not change over the 146-day period of immunization and were not influenced by the nature of the carrier. The implications of these findings in terms of the phylogenetic origins of antibody diversity are discussed.