DEVELOPMENT OF POLYVALENT RECOMBINANT VACCINES FOR CATS.

猫多价重组疫苗的开发。

• 批准号: 07556069
• 负责人: MIKAMI Takeshi
• 金额: $ 12.61万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07556069/
• 关键词: recombinant vaccine; feline herpesvirus 1; feline calicivirus; feline immunodeficiency virus; gag protein; capsid protein; thymidine kinase gene; immunoblot analysis; FHV-1; TK; FCV; 間接蛍光抗体法; イムノブロット解析; FIV; 組換え多価ワクチン; ウイルス性鼻気管炎; 組換えウイルス /

The aim of the present studies is to develop polyvalent recombinant (rec) vaccines which are superior to safeness, inexpensiveness and effctiveness to cats, and to put the vaccines to practical use. The following results are obtained.1.We succeeded in the attenuation of thymidine kinase (TK) deficient mutant (C7301dlTK) of feline herpesvirus type 1 (FHV-1) in cats and the construction of a recombinat FHV-1 (C7301dlTK-Cap) inserted a precursor capsid gene of feline calicivirus (FCV) into the TK deletion locus of the C7301dlTK.Cats vaccinated with C7301dlTK-Cap and then challenged with virulent FHV-1 and FCV were protected to a significant degree against the manifestations of the illness caused by both viruses. However, immune presponse to FCV was weak.2.Therfore to improve further on the recombinat FHV-1, we constructed an improved recombinant FHV-1 (dlTK (gCp) -Cap) carring a putative FHV-1 gC promoter sequence on the upstream of the FCV precursor capsid gene of the C7301dlTK-Cap. Growth kinetics of the dlTK (gCp) -Cap in cell cultures was similar to those of C7301dlTK and C7301dlTK-Cap. A strong expression of FCV immunogenic antigen by dlTK (gCp) -Cap was confirmed by indirect immunofluorescence and enzime-linked immunosorbent assays. In addition, one vaccination with dlTK (gCp) -Cap protected cats more effectively against subsequent virulent FCV challenge than that with C7301dlTK-Cap.3.We constructed a recombinat FHV-1 expressing Gag protein of feline immunodeficiency virus (FIV), in which a cDNA encoding the Gag protein of FIV was inserted at the TK deletion site of C7301ddlTK,and was designated as C7301ddlTK-gag. Growth kinetics of the reconbinat viruses in Crandell feline kidney cells was similar to that of the parent C7301 strain. By immunoblot analysis, C7301ddlTK-gag was confirmed to express the FIV Gag precursor protein in the cells.

本研究的目的是研制对猫安全、廉价、有效的多价重组疫苗，并将其投入实际应用。1.成功地减毒猫疱疹病毒1型(FHV-1)胸苷激酶(TK)缺失突变株(C7301dlTK)，并构建了将猫杯状病毒(FCV)前体衣壳基因插入C7301dlTK缺失位点的重组FHV-1(C7301dlTK-Cap)。用C7301dlTK-Cap免疫猫后，对两种病毒引起的病变均有明显的保护作用。因此，为了进一步改进重组FHV-1，我们在C7301dlTK-Cap的FCV前体衣壳基因上游构建了一个含有FHV-1 GC启动子序列的改良型重组FHV-1(dlTK(GCP)-Cap)。DlTK(GCP)-Cap在细胞培养中的生长动力学与C7301dlTK和C7301dlTK-Cap相似。间接免疫荧光和酶联免疫吸附试验证实dlTK(GCP)-Cap表达FCV免疫原性抗原。此外，与C7301dlTK-Cap.3疫苗相比，dlTK(GCP)-Cap疫苗能更有效地保护猫抵抗后续的FCV攻击。重组病毒在Crandell猫肾细胞中的生长动力学与亲本株C7301相似。免疫印迹分析证实C7301ddlTK-Gag在细胞内表达FIV Gag前体蛋白。

项目成果

Maeda,K.,et.al.: "Expression and properties of feline herpesvirus type 1 gD(hemagglutinin)by a recombinant baculovirus." Virus Research. (In press). (1997)

Maeda,K.,et.al.：“重组杆状病毒表达猫疱疹病毒 1 型 gD（血凝素）的表达和特性。”

Maeda, K. et al.: "Expression and identification of the feline herpesvirus type 1 glycoprotein B(gp143/108)." Virus Res.39. 55-61 (1995)

Maeda, K. 等人：“猫疱疹病毒 1 型糖蛋白 B (gp143/108) 的表达和鉴定。”

Yokoyama, N., et al.: "Recombinant viral vector vaccines for the veterinary use. Review article." J.Vet.Med.Sci.59, (5). 311-322 (1997)

Yokoyama, N. 等人：“兽用重组病毒载体疫苗。评论文章。”

Maeda, K. et al.: "Restriction endonuclease analysis of field isolates of feline herpesvirus type 1 and identification of the heterogeneous regions." J. Clin.. Microbiol.33. 217-221 (1995)

Maeda, K. 等人：“猫疱疹病毒 1 型现场分离株的限制性核酸内切酶分析和异质区域的鉴定。”

Yokoyama, N. et al.: "Pathogenicity and vaccine efficacy of a thymidine kinase-deficient mutant of feline herpesvirus type 1 in cats." Arch.Virol.(In press).

Yokoyama, N. 等人：“猫疱疹病毒 1 型胸苷激酶缺陷突变体对猫的致病性和疫苗功效。”