New B cell epitopes in the Plasmodium falciparum malaria circumsporozoite protein

Abstract

Most antibodies directed against the Plasmodium falciparum circumsporozoite (CS) protein react with its central domain, which contains about 40 repeats of the tetrapeptide Asn‐Ala‐Asn‐Pro (NANP). To search for new epitopes in the non‐repetitive part of the CS protein, we expressed the non‐repetitive regions of the protein in E. coli as fusion proteins with mouse dihydrofolate reductase linked to six adjacent histidine residues. These fusion proteins were obtained at > 70% purity by a single Ni‐chelate affinity chromatography step. Of the new epitopes defined in the C‐terminal portion of the CS protein, three are located in a stretch of 65 amino acids immediately C‐terminal of the protein's central repetitive domain. Pooled sera from inhabitants of a malaria‐endemic area reacted with epitopes in this region of the molecule, and four mouse monoclonal antibodies to this region also reacted with the native CS protein on sporozoites. Two of the monoclonal antibodies reacted with a peptide PNDPNRNVD derived from a conserved region of the CS protein. The other two antibodies showed different reactivities to sporozoites of the NF54 and Ro59 parasite isolates. One, which reacted with a peptide ENANANNAV, recognized Ro59 but not NF54 sporozoites, while the other reacted with a small percentage of NF54 but not Ro59 sporozoites. Antibodies which react with non‐repetitive regions of the CS protein could contribute to maintaining its genetic variability.