Genetic restriction and specificity of the immune response in mice to fusion proteins containing repeated sequences of the Plasmodium falciparum antigen Pf155/RESA.

The genetic restriction and specificity of the immune response in mice to two fusion proteins, ZZ-M3 and ZZ-M5, were studied. These proteins contain two IgG-binding domains (ZZ) from staphylococcal protein A, and repeated sequences from the C-terminal [(VEHDAEEN)5 (VEEN)10] (M3) or central [(VEEPTVADDEH)3(VEEPTVAEEH)2] (M5) regions of the Plasmodium falciparum malaria blood stage antigen Pf155/RESA. Strong antibody and T-cell responses to M3 and M5 were linked to expression of the I-Ak allele, and T-cell responses to the bacterial fusion partner ZZ were restricted to mice of the H-2k haplotype. The response to M5 was less restricted than that to M3, giving intermediate responses in mice of H-2d haplotypes as well. However, ZZ-M5-primed lymph node (LN) cells from these mice were primarily induced to proliferate in vitro by the complete ZZ-M5 construct and not by synthetic peptides representing the repeated subunits in M5. The reactivity with intact Pf155/RESA in erythrocyte membrane immunofluorescence was weak of antisera from mice immunized with ZZ-M5, whereas the reactivity of antisera from mice immunized with ZZ-M3 roughly paralleled their reactivity with M3 in an enzyme-linked immunosorbent assay (ELISA). The antibody responses induced by immunization with ZZ-M3 or ZZ-M5 were specific for M3 or M5, respectively, while activated T cells displayed cross-reactivity between M3 and M5 in an in vitro proliferation assay. The results indicate that the assembly of repeated sequences in fusion proteins affects both the MHC class II restriction and the specificity of the induced antibody and T-cell responses.