tailieunhanh - báo cáo hóa học:" Recombinant HPV16 E7 assembled into particles induces an immune response and specific tumour protection administered without adjuvant in an animal model"

Tuyển tập các báo cáo nghiên cứu về hóa học được đăng trên tạp chí sinh học đề tài : Recombinant HPV16 E7 assembled into particles induces an immune response and specific tumour protection administered without adjuvant in an animal model | Petrone et al. Journal of Translational Medicine 2011 9 69 http content 9 1 69 JOURNAL OF TRANSLATIONAL MEDICINE RESEARCH Open Access Recombinant HPV16 E7 assembled into particles induces an immune response and specific tumour protection administered without adjuvant in an animal model 1 2 1 -3 2 1 Linda Petrone Maria G Ammendolia Armando Cesolini Stefano Caimi Fabiana Superti Colomba Giorgi and Paola Di Bonito1 Abstract Background The HPV16 E7 protein is both a tumour-specific and a tumour-rejection antigen the ideal target for developing therapeutic vaccines for the treatment of HPV16-associated cancer and its precursor lesions. E7 which plays a key role in virus-associated carcinogenesis contains 98 amino acids and has two finger-type structures which bind a Zn ion. The ability of an Escherichia coli-produced E7-preparation assembled into particles to induce protective immunity against a HPV16-related tumour in the TC-1-C57BL 6 mouse tumour model was evaluated. Methods E7 was expressed in E. coli purified via a one-step denaturing protocol and prepared as a soluble suspension state after dialysis in native buffer. The presence in the E7 preparation of particulate forms was analysed by non-reducing SDS-PAGE and negative staining electron microscopy EM . The Zn ion content was analysed by mass-spectrometry. Ten pg of protein per mouse was administered to groups of animals once twice or three times without adjuvant. The E7-specific humoral response was monitored in mice sera using an E7-based ELISA while the cell-mediated immune response was analysed in mice splenocytes with lymphoproliferation and IFN-y ELISPOT assays. The E7 immunized mice were challenged with TC-1 tumour cells and the tumour growth monitored for two months. Results In western blot analysis E7 appears in multimers and high molecular mass oligomers. The EM micrographs show the protein dispersed as aggregates of different shape and size. The protein appears clustered