| Co-Investigator(Kenkyū-buntansha) |
HATA Hidekazu Chiba University School of Medicine, Research Associate, 医学部, 助手 (00110304)
NOROSE Kazumi Chiba University School of Medicine, Research Associate, 医学部, 助手 (30156244)
YANO Akihiko Chiba University School of Medicine, Professor, 医学部, 教授 (20135122)
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| Research Abstract |
Molrecular chaperon HSP70 has been revealed to be an antigen presenting molecule of Toxoplasma gondii (T.gondii)-infected cells. We have carried out the cloning of T.gondii-derived HSP70 and have analysed that T.gondii-infected mice recognized T.gondii HSP70 (T.g.HSP70) as a B cell epitope instead of the existence of high homology between T.g.HSP70 and mouse HSP70. Based on these results, we have investigated the roles of T.g.HSP70 and T.g.HSP30/bag1 in the prophylactic immunity in mice against T.gondii infection (IXth International Congress of Parasitology, Monduzzi Editore, 457, 1998 ; Jpn.J.Trop.Med.Hyg.26 : 305, 1998 ; Microbiol. Immunol.43 : 471, 1999 : Cell Stress ＆ Chaperones, 5 (4) : 328, 2000).
By using SAG1 (a major surface antigen of T.gondii) genes-transfected murine antigen presenting cell (APC) lines, we already reported the induction of protective immunity. In this research, we have evaluated the vaccine effects of in vivo gene-vaccinated skingraft against T.gondii infect
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ion and have analyzed the effect of the direct gene vaccination to professional APC, i.e. langerhans cells (LC) and dendritic cells (DC) in the induction of protective immunity. We have analyzed that the successful gene vaccination to LC/DC could be carried out with a shot at 400psi discharge pressure by a gene gun. By using the gene gun, cDNA coding T.gondii SAG1 was intracutaneously vaccinated into C57BL/6 (B6)(a susceptible strain), BALB/c (a resistant strain) and (C57BL/6×BALB/c) F1 (CBF1) mice, and the gene-vaccinated skins of these strains were individually transplanted to CBF1 mice. In antibody production against SAG1, gene-vaccinated B6 skingrafted CBF1 mice were high responder whereas both BALB/c skingrafted CBF1 mice and CBF1 skingrafted CBF1 mice were low responders. Analytical flow cytometry was carried out and the data showed that donor-derived LC/DC from the skingraft migrated to draining lymph nodes of the recipients within 3 days. The effective vaccination was developed in CBF1 mice which received skingraft of the SAG1 gen-vaccinated BALB/c mice against T.gondii challenge infection. These results indicated that the new procedure of in vivo gene-vaccinated skin transplantation for overwhelming the genetic low responder host was established and the gene-vaccination to LC/DC of B6 and BALB/c mice induced different immune responses in CBF1 mice. Less
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