Chaperon gene vaccination for Toxoplasmosis

分子伴侣HSP70是弓形虫感染细胞的抗原提呈分子。我们进行了弓形虫来源的HSP70的克隆，并分析了弓形虫感染的小鼠识别弓形虫HSP70(T.g.HSP70)为B细胞表位，而不是T.g.HSP70与小鼠HSP70存在高度同源性。基于这些结果，我们研究了T.g.HSP70和T.g.HSP30/bag1在小鼠对弓形虫感染的预防性免疫中的作用(IX国际寄生虫学大会，Monduzzi Editore，457,1998；Jpn.J.Trop.Med.Hyg.26：305,1998；Microbiol.免疫学.43：471,1999：cell Stress&Chaperones，5(4)：328,2000)。通过使用SAG1(弓形虫的主要表面抗原)基因转染的小鼠抗原提呈细胞(APC)，我们已经报道了诱导保护性免疫的报道。在本研究中，我们评价了体内基因疫苗对弓形虫感染…的疫苗效果。分析了直接基因疫苗对专业性APC，即朗格汉斯细胞(LC)和树突状细胞(DC)诱导保护性免疫的作用。我们分析了用基因枪在400psi放电压力下进行一次成功的LC/DC基因疫苗接种。用基因枪将弓形虫SAG1基因分别皮内接种于C57BL/6(B6)(敏感株)、BALB/c(抗性株)和(C57BL/6×BALB/c)F1(CBF1)小鼠，并将接种后的皮肤移植到CBF1小鼠体内。在抗SAG1抗体的产生上，接种B6基因的CBF1小鼠表现为高应答，而BALB/c接种CBF1小鼠和CBF1接种CBF1小鼠均为低应答。流式细胞术分析结果显示，供者来源的LC/DC在3d内迁移至受者的引流淋巴结。用SAG1基因免疫BALB/c小鼠的SKINGRAFT对弓形虫攻击感染的CBF1小鼠进行了有效免疫。这些结果表明，建立了体内基因免疫皮肤移植以克服遗传低应答宿主的新程序，B6和BALB/c小鼠的LC/DC基因疫苗在CBF1小鼠中诱导了不同的免疫应答。较少

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 … More 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