Identification of the Pulmonary Stem/Progenitor Cells From Different Species as the Target Cells for Influenza Virus Infection

• Main Authors: Yu-Ming Lin, 林佑名
• Other Authors: Thai-Yen Ling
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/10589782372852054497

碩士 === 國立臺灣大學 === 藥理學研究所 === 96 === We have reported a serum-free primary culture system to generate pulmonary epithelium stem/progenitor cells (Oct-4+/SSEA-1+/Sca-1+ and CCSP+) with surrounding stroma cells in mouse. These stem/progenitor cells were the target cells for SARS-CoV infection in primary cultures. However, the targeting potential of these cells for other lung infection-associated viruses remains unclear. We demonstrated the susceptible ability of these colony cells for influenza virus infection, such as H3N2 and H5N1.After H3N2 infection, HMGB1(High mobility group box 1 ) predominantly translocated to the cytoplasm in colony cells, and the pro-inflammatory cytokines like IL-1β, IL-6 TNF-α were up-regulated. However, H5N2 virus could not infect the cells at all. So far, the available cell line to study influenza virus infection was MDCK, which derived from kidney cells of dog. To examine that the pulmonary epithelium stem/progenitor cells were the first target for virus infection, and study the cross-species infection of influenza virus, we established both chicken and mouse pulmonary stem/progenitor cells and to compare their susceptibility of virus infection. The chicken lung epithelial cells were susceptible for H5N1, H5N2, H6N1 and H7N7 virus. Further experiments by RT-PCR and Q-PCR showed the expression of stem cell markers in the chicken lung cells, including alkaline phosphatase (cAP), cPouV, cSOX2, but not cNanog. The E-cadherin, pulmonary surfactant protein, and telomerase were also demonstrated to be expressed in cells. These observation suggested that the chicken pulmonary stem/progenitor cells are able to serve as a good cell model to study the influenza virus infection. Furthermore, we transfected the chicken pulmonary epithelial cells with plasmid pRSV-TEX to establish a cell line suitable for cell-based vaccine production. These cells were subcultured to passage 6, and expressed the same marker as primary cells. SV40 large T antigen also can be detected in passage 6 cells.