Ehime Univ   Graduate School of Medicine  Mol Parasitol   Graduate Course

How to become a foreign student of Ehime University (International student center) Please feel free to contact with us (torii@m.ehime-u.ac.jp)

Guide for the Graduate Course (Year 2006) Key Word :: Technique :: Research Theme 1) Key Word Malaria (Plasmodium falciparum, vivax, yoelii, berghei) Erythrocyte Invasion mechanism Mosquito Transmission-Blocking Vaccine 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. Luciferase assay, and so on. Immunological Technique     Polyclonal & monoclonal antibody production, Flow Cytometry, ELISA, SDS-PAGE, Western Blot     Immunoprecipitation,IFA, and so on. Immunoelectron Microscopy Cell Culture     Plasmodium falciparum (Transfection work is going on), COS cell, CHO cell Technique to maintain Parasite     Plasmodium yoelii, Plasmodium berghei with mouse Technique to maintain Mosquito     Anopheles stephensi 3) Research Theme      We are focusing on the basic research for malaria parasite, Plasmodium, and conducting P. vivax transmission blocking vaccine development. A) Molecular and Cell Biology of the Erythrocyte Invasion by Plasmodium merozoite (Kaneko)      Regarding to the failures of some 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.      We have identified 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 we developed a novel erythrocyte binding assay using Flowcytometer and found that GPI-anchor is involved of the P. yoelii RhopH complex binding to the erythrocyte (Rungruang et al. 2005).  We are currently conducting to identify the receptor(s), and elucidate a function of the RhopH complex using advanced techniques including gene-targeting.      We are also interested in identifying unknown receptors for the other invasion-related molecules including proteins encoded by ebl (erythrocyte-binding-like) and rbl (reticulocyte-binding-like) families.  Join us and together let's find them all.  We are now in a very good shape. B) Transmission Blocking Vaccine Development (Torii)      We are studying the novel surface proteins of ookinete and gametocyte using molecular biological technique, including general procedure for the gene cloning, sequencing, recombinant protein expression. and so on.  In the past few years, we cloned P. vivax 25kDa and 28kDa ookinete surface proteins (Pvs25 and Pvs28, respectively) and showed that antibodies against these proteins are capable to block transmission by the mosquitoes.  Especially, transmission was blocked with only antibodies against Pvs25, which indicates that this molecule is a strong candidate for the transmission blocking vaccine. We are conducting the field experiment of P. vivax transmission blocking vaccine in collaboration with National Institute of Health, USA and AFRIMS, Thailand.

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