Department of Protozoology
 Nagasaki Univ   Institute of Tropical Medicine Department of Protozoology Graduate Course

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Key Word :: Technique :: Research Theme :: Graduate School of Biomedical Sciences

1) Key Word

Malaria (Plasmodium falciparum, vivax, yoelii)
     Erythrocyte Invasion mechanism
     Function of the Malaria-encoded molecules expressed on the erythrocyte surface

2) Techniques we are using in our laboratory

Molecular Biology
    Gene Cloning, Sequencing, PCR, real-time PCR, Southern, Northern,
    Recombinant protein expression (E. coli and mammalian cells).
    Gene transfection to P. falciparum and P. yoelii Luciferase assay, and so on.
Immunological Technique
    Polyclonal antibody production, Flow Cytometry, ELISA, SDS-PAGE, Western Blot
    Immunoprecipitation, IFA, and so on.
Cell Culture
    Plasmodium falciparum and P. knowlesi (Transfection), COS cell, CHO cell
Technique to maintain Parasites
    Rodent malaria parasites such as P. yoelii and P. berghei using mouse

3) Research Theme

     We are focusing on the basic research for malaria parasite, Plasmodium, and assessing their potential use for a vaccine development using malaria field isolates.

A) Molecular and Cell Biology of the Erythrocyte Invasion by Plasmodium merozoite

     Regarding to the unsuccessful vaccine trails against malaria, we believe that the further understanding of the parasite biology is essential to develop the effective vaccine.  Since merozoite is the only stage that is exposed to the host immune system during blood stage, the molecules expressed at the merozoite stage are the good vaccine targets.  The merozoite stage parasite invade erythrocytes in specific manner, in other words, merozoite recognize specific receptors on the surface of the erythrocytes.  But little is known about the precise host-parasite interaction at the molecular level.
     Kaneko was involved in the project conducted in Ehime University, to identify genes encoding rhoptry protein, the RhopH complex (Kaneko et al. 2001; Ling et al. 2003), and showed that three of the component RhopH1 were encoded by members of clag multigene family (Ling et al. 2004; Kaneko et al. 2005).  Furthermore Rungruang and Kaneko developed a novel erythrocyte binding assay using Flowcytometer and found that GPI-anchor was involved of the P. yoelii RhopH complex binding to the erythrocyte (Rungruang et al. 2005).  Also in collaboration with a group of Karolinska Institutet, Sweden, Kaneko found a novel surf multigene family and an extracellular DBL-like domain of Pf332 protein expressed on the malaria infected-erythrocytes (Winter et al. 2005; Moll et al. 2007). Recently our group including Culleton and Kaneko identified EBL is a critical virulence determinant between P. yoelii virulent and avirulent lines (Pattaradilokrat et al, 2009; Otsuki et al, 2009; Li et al, 2011).

     On May/1/2007, Kaneko joined to Nagasaki University, Institute of Tropical Medicine to conduct Department of Protozoology.  New projects are starting in Nagasaki related to the molecules involved in the erythrocyte invasion, in addition to the continued project to elucidate a function of the RhopH complex using advanced techniques including gene-targeting.  Those are including analysis of the protein sorting signal of the invasion-related proteins, identification of the novel erythrocyte binding protein/domain, and so on.

B) Assessment of the selective pressure on the malaria antigens

     When we find molecules involved in the erythrocyte invasion or showing host cell binding activity, we analyze nucleotide sequence diversity using malaria isolates collected from the endemic areas and assess the host selective pressure on the proteins to evaluate its potential usage as a vaccine.  We are collaborating researchers in many malaria endemic areas, such as Thailand, China, Laos, Brazil, and so on.

4) Graduate School of Biomedical Sciences [Link]

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