| Co-Investigator(Kenkyū-buntansha) |
ITO Akira Asahikawa Med.Col., Facul.of Medicine, Prof., 医学部, 教授 (70054020)
SHIBAHARA Toshiyuki Tottori University, Facul.of Medicine, Assoc.Prof, 医学部, 助教授 (70116937)
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| Research Abstract |
Cysticercosis, one of the most serious larval cestodiasis of medical importance in humans and of economic importance in pigs, is spreading worldwide, especially in Asia, Africa and Latin America. The trial of the vaccine development against Taenia solium cysticercosis has been made in many countries, and actually some candidate molecules have been also reported. However, the effects of vaccine used those molecules have been not always perfect. Expression-library immunization has been, proposed as an effective means to screen a large number of genes of the pathogen as candidate protective molecules. In this study, we examined the efficacy of expression-library immunization for vaccine development against T.solium cysticercosis using T.taeniaeformis-rat model system
Total RNAs were isolated from last 10 segments of adult T.taeniaeformis by Isogen LS, and poly A RNA was purified using oligotex-dT30 (Takara). cDNA library was produced using SuperScript Plasmid System (Inbitorogen), which contain a mammalian expression vector, pCMV・SPORT6. Transformed E.coli were plated onto LB-plate containing ampicillin, and the cells that grew on the plate were resuspended in TYGPN nedium. Each clone was sequenced and classified with the property of the insert. Twenty to thirty independent clones were simultaneously propagated in LB medium with ampicillin, and plasmid DNAs were isolated and purified using End Free Plasmid Kit. (QIAGEN). For the immunization, purified plasmid DNAs were injected directly into the spleen and muscle of mice
It was reported that the number of the mitochondrion in adult taeniid cestode was not so many. However, more than 30% of the sequenced clones had mitochondrial genes. Specific antibody to Taenia taeniaeformis was detected by western-blot in the mouse immunized with Library-Vaccine
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