The effect of specific cell inactivation on the cellular immune response to ferredoxin peptides

Previous studies using synthetic immunogenic molecules containing two haptenic peptides (ther NH2-terminal heptapeptide and the COOH-terminal pentapeptide of oxidized ferredoxin (O-Fd) from C. pasteurianum) have shown that both peptides individually are capable of initiating lymphocyte transformation and inhibiting migration in populations of lymphocytes from O-Fd-sensitized guinea pigs. While migration inhibition could readily be stimulated by single haptenic peptides, lymphocyte treasformation appeared to be more easily induced by molecules containing two or more haptenic peptides (these could be identical or different) (Kelly, B., Levy, J.G. and Hull, D., Eur. J. Immunol., 1973. 3:574). If lymphocyte transformation is a T cell-mediated phenomenon, these observations indicate the possibility of T cell-T cell interaction. The two haptenic peptides (designated "N" and "C") were synthesized and conjugated to succinylated bovine serum albumin (S-BSA), forming the conjugates N-S-BSA and C-S-BSA, respectively. These conjugates were labeled to high specific activity with 125iodine and were used in an "antigen suicide" procedure to treat guinea pig lymph node cell preparations previously sensitized to O-Fd and keyhole limpet hemocyanin (KLH). Cell populations exposed to either 125I-labeled C-S-BCA demonstrated decreased lymphocyte transformation in the presence of O-Fd but not in the presence of KLH. These results indicate specific cell inactivation by the radioactive peptide conjugates of those cells responsible for initiating cell transformation. Experiments performed by mixing 125I-labeled N-S-BSA-treated cells with 125I-labeled C-S-BSA-treated cells were successful in partially restoring the response to O-Fd and suggest possible synergy between N and C determinant binding cells in the cellular immune response to O-Fd. Evidence from B cell depletion studies suggests that this is a T cell-T cell interaction.