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Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
Production and verification of human papillomavirus type 18 vaccine in vitro
Shujie Liao1, Shixuan Wang, Ling Xu
1Cancer Biology Research Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Hubei 430030, P.R. China.
Abstract:
This study aimed to generate human papillomavirus (HPV) type 18 L1 virus-like particles (VLPs) and investigated the immunity effectiveness in vitro by using the Bac to Bac baculovirus expression system and Ni-NTA purification system to express and purify the VLP. The bio-function and immune response of purified VLP were analyzed by a mouse erythrocyte haemagglutination assay and the IFN-gamma ELISPOT. The proteins of interest, primarily located at the nucleus, formed VLP by self-assembly, and had an effective immune activity in vitro. The level of IFN-gamma in the specific HPV18 positive group was significantly higher than that of the mixed high-risk HPV infected group and non-HPV infection one in vitro. Our study indicated that HPV18 later protein L1 was efficiently expressed in Bac to Bac baculovirus expression system and the HPV18 L1-VLPs showed immunoreactivity and bio-activity in vitro. Furthermore, the ELISPOT results support the hypothesis that the cell-mediated immunity generated by HPV18 L1-VLP may cross-type among high-risk HPV types.

