Research in mechanism of attachment and penetration of EB virus to the epithelial cell.
Grant-in-Aid for Scientific Research (B)
|Allocation Type||Single-year Grants |
|Research Institution||HOKKAIDO UNIVERSITY |
TAKADA Kenzo Hokkaido Univ., Inst. for Gen. Med., Prof., 遺伝子病制御研究所, 教授 (30133721)
YOSHIYAMA Hironori Hokkaido Univ., Inst. for Gen. Med., Asso. Prof., 遺伝子病制御研究所, 助教授 (10253147)
SETO Eri Hokkaido Univ., Inst. for Gen. Med., Inst., 遺伝子病制御研究所, 学術研究員 (40431382)
丸尾 聖爾 北海道大学, 遺伝子病制御研究所, 助教授 (70292018)
南谷 武春 北海道大学, 遺伝子病制御研究所, 学術研究員 (00374679)
|Project Period (FY)
2005 – 2006
Completed(Fiscal Year 2006)
|Budget Amount *help
¥14,600,000 (Direct Cost : ¥14,600,000)
Fiscal Year 2006 : ¥6,300,000 (Direct Cost : ¥6,300,000)
Fiscal Year 2005 : ¥8,300,000 (Direct Cost : ¥8,300,000)
|Keywords||EBV / Epithelial cell / Receptor / Ligand / Attachment|
1.Research in EBV Receptor in Epithelial Cell
Epstein-Barr Virus (EBV) infects both B lymphocytes and epithelial cells. EBV uses CD21 as a receptor and gp350 as a viral ligand for its adsorption to B lymphocytes. However, epithelial cell dose not or scarcely express CD21 and the mechanism how EBV infects to epithelial cells is not known. We showed that CD21-negative epithelial cell was susceptible to EBV infection. We have also showed that pretreatment of epithelial cells with anti-CD21 antibody or soluble form of CD21 prior to EBV infection could not block the infection. These facts suggest that EBV infects epithelial cells using an alternative receptor other than CD21.
AGS cell is a human gastric epithelial cell which is highly susceptible to EBV infection, but dose not express CD21. Retroviral cDNA library was constructed from mRNA extracted from AGS cells in order to isolate a gene of the epithelial EBV receptor using the method of expression cloning.
Canine MDCK cell and human HeLa c
ell are cells which are originally resistant to EBV infection. However, once CD21 gene is introduced to these cells and CD21 molecules are expressed on the cells, these cells become susceptible to EBV infection. Because of this feature, MDCK cell and HeLa cell can be usedto isolate a novel gene of the EBV receptor by expressing cDNA. We have expressed cDNA on MDCK cells and isolated several candidate genes of a novel EBV receptor. Among them, a gene was isolated repeatedly by the independent experiments. We are going to introduce and express the candidate gene to HeLa cells and will examine the susceptibility to EBV infection.
2. Research in EBV Ligand to the Epithelial Cell Receptor
It has been considered that the viral ligand for EBV infection on epithelial cells was the gp85-gp25 complex. However, we have found that the gp85 knockout EBV could infect to epithelial cells with a frequency 10,000 times lower than the frequency of the wild type EBV. We have also found that the epithelial cell infected with gp85 knockout EBV could produce infectious EBV. The produced gp85 knockout EBV could infect B lymphocytes and transform the B lymphocyte to lymphoblastic cell lymphocyte (LCL). It is suggested that EBV can infect both epithelial cells and B lymphocytes by the mechanism which dose not use gp85. Less
Research Products (40results)