The primary focus of the project was CTL mediated responses to the hsp 70 of Plasmodium falciparum. The work involved cloning and expression of the P.falciparum hsp 70 gene is a Salmonella system. The gene was also cloned into vectors for use in DNA immunization of different murine strains and in the selectable mammalian expression vector pcDNA3 (Invitrogen). 

 

Experiments were initiated to evaluate the expression of the gene in different mammalian cell lines prior to the immunization of the different murine strains with recombinant DNA vector constructs and the establishment of stable transfectants. Peptides were also designed based on predictive analysis of the P.falciparum hsp 70 gene with reference to CTL epitopes of different murine haplotypes. Experiments were initiated to standardize the CTL assays in different strains of mice using various preexisting vaccinia constructs expressing hsp 70. 

 

In a separate project the P.falciparum hsp 60 gene was expressed in E.coli and the protein partially purified using a 5' polyhistidine tag. Purified protein was used to produce antibodies in mice and experiments were performed to evaluate the expression of hsp 60 in P.falciparum and to evaluate the distribution and localization of hsp 60 in different parasite stages.