We believe that the approach we have utilized contributes to more complete understanding of the role of HLA class II molecules in presentation of antigen to T lymphocytes and helps to resolve paradoxes presented by earlier studies in which class II-associated restrictions were not apparent. The approach has depended upon development of a library of cloned TCL reactive to a particular haptenic determinant in various class II MHC contexts. It employed initial screening for MHC restriction by analysis on allogeneic stimulator cell panels that could be confirmed or clarified by antibody blocking experiments employing mAb of defined specificities. This has led to clear identification of TCL restricted to recognition of TNP in the context of each of the major class II molecular species, DR, DQ and DP. It has also raised the intriguing and novel possibility that in some instances, at least for DP, T-cells may employ restricting elements, as revealed by mAb blocking, that are either nonpolymorphic or of very low polymorphism. Finally, where important functional polymorphisms have been identified at apparent variance from defined serological specificities, it has been possible to correlate antigen recognition with molecular polymorphisms defined at both the protein level, by amino acid sequencing, and the genomic level by restriction enzyme polymorphisms. As these approaches are combined with new molecular genetic techniques for generation of novel class II constructs, it seems likely that important, but as yet unresolved, questions relating to the nature of the ternary complex involving antigen, MHC molecule and T-cell receptor will be resolved.