Characterization of a differential immunoscreen epitope of Plasmodium falciparum using combinatorial agents.
A differential serological screening of a lambdagt11 cDNA expression library has identified several clones, which react exclusively to sera samples from persons clinically immune to malaria but not to acute malaria patient sera. One such clone, IPf9, has a 315-bp cDNA insert, which was found to be conserved in different strains of the human and rodent malarial parasite Plasmodium falciparum and Plasmodium berghei, respectively. The induced expression product of IPf9 was used to generate polyclonal sera in rabbits. The IPf9 expression product was also screened with phage surface display combinatorial libraries to isolate reagents that specifically bound to the IPf9 product. The polyclonal antisera and the combinatorial reagents recognized a 50-kDa protein from P. falciparum, and a 53-kDa product from P. berghei. Immunofluorescence studies using asexual and sexual stages of P. falciparum showed the protein to be present within the parasite in each of the asexual and sexual stages. The combinatorial reagents showed a partial inhibition in the growth of P. falciparum in vitro. Mice infected with the P. berghei showed the presence of T-cells that exhibited lymphoproliferation when stimulated with the IPf9 protein. It is suggested that IPf9 protein is a conserved protein epitope, and may be relevant for a protective immune response to malaria.
Singh
N J
NJ
Department of Biological Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Mumbai 400 005, India.
Sehgal
A
A
Sharma
S
S
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D020650:Combinatorial Chemistry Techniques; D018604:Epitope Mapping; D006801:Humans; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D010962:Plasmodium berghei / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D010963:Plasmodium falciparum / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D011487:Protein Conformation; D011817:Rabbits
D000953:Antigens, Protozoan; C414223:IPf9 protein, Plasmodium berghei