| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2004: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Research Abstract |
A combinatorial human immunoglobulin gene library was constructed from peripheral lymphocytes of eight patients with falciparum malaria. The library was screened for the production of Fab antibody fragments to Plasmodium falciparum. Three Fab clones, Pf25, Pf143 and Pf227, reacted with recombinant 19-kDa C-terminus fragment of P. falciparum merozoite surface protein-1 (MSP-1_<19>) in enzyme-linked immunosorbent assay. Indirect immunofluorescent microscopy revealed that these Fabs reacted with the surface of merozoites in schizont stage of the parasite. The closest heavy-chain germlines were VH1-8,D3-10 and JH4 (Pf25 and Pf227), or VH7-81,D3-10 and JH5 (Pfl43). On the other hand, the closest light-chain germlines were A27 and Jκ5 (Pf25), A27 and Jκ2 (Pfl43), or A27 and Jκ4 (Pf227). The dissociation constants of Pf25,Pf143 and Pf227 were 1.09×10^<-9>M,1.71×10^<-9>M and 2.66×10^<-9>M, respectively. When Pf25 was added to cultures of P. falciparum at the concentration of 0.2 mg/ml, no inhibitory effect was observed on the growth of the parasite. However, the immunoglobulin gene library prepared from patients seems to be a potent tool in the production of human antibodies for clinical diagnosis, immunoprophylaxis and treatment of falciparum malaria.
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