Project/Area Number |
12460133
|
Research Category |
Grant-in-Aid for Scientific Research (B)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Basic veterinary science/Basic zootechnical science
|
Research Institution | THE UNIVERSITY OF TOKYO |
Principal Investigator |
OTSUKA Haruki THE UNIVERSITY OF TOKYO, Graduate School of Agricultural and Life Sciences, Professor (80261957)
|
Co-Investigator(Kenkyū-buntansha) |
MATSUMOTO Yasunobu THE UNIVERSITY OF TOKYO, Graduate School of Agricultural and Life Sciences, Assistant (90251420)
TAKASHIMA Yasuhiro THE UNIVERSITY OF TOKYO, Graduate School of Agricultural and Life Sciences, Assistant (20333552)
松本 芳嗣 東京大学, 大学院・農学生命科学研究科, 助教授 (00173922)
|
Project Period (FY) |
2000 – 2002
|
Project Status |
Completed (Fiscal Year 2002)
|
Budget Amount *help |
¥13,600,000 (Direct Cost: ¥13,600,000)
Fiscal Year 2002: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥6,500,000 (Direct Cost: ¥6,500,000)
|
Keywords | BHV-1 / PRV / TF7-6 / Th1 / apoptosis / UsORF8 / D8 / RK13 / semi-permissive / non-permissive / GFP / Th2 / gB / gC / HmLu-1 / IE gene / green fluorescent protein / A31細胞 |
Research Abstract |
Bovine herpesvirus 1 (BHV-1) has a relatively narrow host cell range in vitro and in vivo when compared to pseudorabies virus (PRV). The aim of this study was to elucidate whether homologous glycoproteins gC and gB from PrV can function in a heterologous BHV-1 background and whether the expression of these PRV glycoproteins influences the in vitro host cell specificity of BHV-1. We constructed BHV-1 recombinant, BHV-1/TF7-6 in which PRV gC and gB were expressed and examined their attachment and penetration properties in permissive, semi-permissive and non-permissive cells. It was found that the expression of PRV gC and gB in a BHV-1 environment facilitated virus attachment and penetration by BHV-1 to semi- or non-permissive cells. When BALB/c mice were inoculated with purified BHV-1/TF7-6 or the parental BHV-1, the former induced lower level of anti-BHV-1 IgG than the latter did. However, the IgG2a:IgG1 ratio was higher in BHV-1/TF7-6 inoculated mice than in the parental BHV-1 inoculated ones. These results indicate that BHV-1/TF7-6 induced type 1 predominant immunity to BALB/c mice. BHV-1 Us ORF8 protein with homology to the aUs9 protein of other alphaherpesviruses induces apoptosis in rabbit kidney (RK13) cells without the presence of other BHV-1-encoded proteins. A mutant of BHV-1 that fails to express BHV-1 UsORF8 protein, BHV-1/D8 exhibited a reduced cyto-toxicity to RK13 cells when compared to the cyto-toxicity of the parental BHV-1. In RK13 cells, the onset of apoptosis was not observed during the infection with BHV-1/D8, and the virus replication of BHV-1/D8 was markedly greater than that of control virus. These date appeared to indicate that the Us ORF8 protein activates the apoptosis process and facilitates virus release from BHV-1 infected cells.
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