How to select Plasmodium falciparum pre-erythrocytic antigens in an expression library without defined probe.
The restricted access to Plasmodium falciparum liver stages has greatly limited the analysis of the antigenic content of that stage. Due to the lack of material to perform immunochemical studies, of access to mRNA, and of monoclonal probes, we decided to screen a genomic library with stage-restricted human antibodies. This strategy led to the identification of a large number of DNA fragments encoding both sporozoite specific as well as liver-stage specific epitopes. Following the initial characterization of one liver-stage antigen, further screening was performed by using additional selective human antibodies. These were defined as having a high degree of reactivity with native antigens on either of the two stages while being negative with the already known molecules of the two stages. From this second screening and the study of cross-reactions, several subsets of DNA clones expressing antigens present on the surface of sporozoites, or in liver stages, or in both, could be identified. In exposed individuals a high prevalence of antibodies to several of these antigens was found.
Marchand
C
C
Parasitologie Biomédicale, Institut Pasteur, Paris, France.
Druilhe
P
P
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D003429:Cross Reactions; D004274:DNA, Recombinant / Q000276:immunology; D000939:Epitopes / Q000235:genetics; D015698:Genomic Library; D006801:Humans; D008099:Liver / Q000469:parasitology; D009693:Nucleic Acid Hybridization; D010963:Plasmodium falciparum / Q000276:immunology*
D000953:Antigens, Protozoan; D004274:DNA, Recombinant; D000939:Epitopes