Grant-in-Aid for Scientific Research (A)
|Allocation Type||Single-year Grants |
Basic veterinary science/Basic zootechnical science
|Research Institution||Nihon University (2001-2002)|
Obihiro University of Agriculture and Veterinary Medicine (1999-2000)
MIKAMI Takeshi Nihon University, Professor, 生物資源科学部, 教授 (20091506)
KABEYA Hidenori Nihon University, Assistant Professor, 生物資源科学部, 助手 (10318389)
MARUYAMA Soichi Nihon University, Associate Professor, 生物資源科学部, 助教授 (30181829)
SAKAI Takeo Nihon University, Professor, 生物資源科学部, 教授 (50147667)
XUAN Xuenan Obihiro University, Associate Professor, 原虫病研究センター, 助教授 (10292096)
NAGASAWA Hideyuki Obihiro University, Professor, 原虫病研究センター, 教授 (60172524)
五十嵐 郁男 帯広畜産大学, 原虫病研究センター, 教授 (80159582)
宮沢 考幸 (宮沢 孝幸) 東京大学, 農学部, 助手 (80282705)
高橋 英司 東京大学, 農学部, 教授 (50183439)
|Project Period (FY)
1999 – 2002
Completed (Fiscal Year 2002)
|Budget Amount *help
¥42,300,000 (Direct Cost: ¥36,900,000、Indirect Cost: ¥5,400,000)
Fiscal Year 2002: ¥11,830,000 (Direct Cost: ¥9,100,000、Indirect Cost: ¥2,730,000)
Fiscal Year 2001: ¥11,570,000 (Direct Cost: ¥8,900,000、Indirect Cost: ¥2,670,000)
Fiscal Year 2000: ¥8,900,000 (Direct Cost: ¥8,900,000)
Fiscal Year 1999: ¥10,000,000 (Direct Cost: ¥10,000,000)
|Keywords||T. gondii / IL-12 / FHV-1 / recombinant vaccine / TgSAG1 / TgSAG2 / TgROP2 / TgSRS2 / T.gondii / Th1細胞 / サイトカイン / ウイルスベクター / FIV / ネコIFN-γ / ネコIL-12 / ネコIFN-Υ|
1) Cloning and expression of feline IL-12 gene
It is known that IL-12 enhances NK activity, stimulates CTL activity and induces INF-γ production, and therefore IL-12 is considered as a key cytokine in Th1 helper cell-derived immune responses. A full length cDNA of feline IL-12 p35 and p40 subunits was cloned, expressed and characterized. The recombinant feline IL-12 has both INF-γ -inducing activity and CTL-stimulating activity.
2) Cloning and expression of the immunodominant antigen genes of T. gondii
The genes encoding immunodominant antigens, P22 (SAG2), P30 (SAG1), P54 (ROP2), and SRS1 of T. gondii were PCR-amplified using cDNA from RH strain as templates, and expressed in E. coli cells. Mice immunized with recombinant antigens produced a partial protective immunity against T. gondii challenge infections.
3) Construction of recombinant feline herpesviruses expressing the antigen genes of T. gondii
Recombinant feline herpesvirus type 1 (FHV-1) expressing immunogenic antigens (P22, P30, P54, SRS1) of T. gondii were constructed, and demonstrated that the recombinant antigens were similar to the native antigens from T. gondii in structure and antigenicity. Construction of FHV-1 expressing IL-12 is under way.
4) Protective effects of recombinant FHV-1 expressing T. gondii antigens against challenge infection in cats
Cats immunized with FHV-1/TgP22, FHV-1/TgP30, FHV-1/TgP54, and FHV-1/SRS1 produced a partial protective immunity against T. gondii infections, respectively. Our next step will be immunization trials with cats to check the potency of the FHV-1/IL-12 to induce effective immunity against toxoplasmosis.