TAILIEUCHUNG - Báo cáo sinh học: "Comparative analysis of macrophage associated vectors for use in genetic vaccine"

Tuyển tập các báo cáo nghiên cứu về sinh học được đăng trên tạp chí sinh học Journal of Biology đề tài: Comparative analysis of macrophage associated vectors for use in genetic vaccine | Ahsan and Gore Genetic Vaccines and Therapy 2011 9 10 http content 9 1 10 GENETIC VACCINES AND THERAPY RESEARCH Open Access Comparative analysis of macrophage associated vectors for use in genetic vaccine Mohammad Feraz Ahsan and Milind M Gore Abstract Background Antigen presentation by non professional antigen presenting cells APC can lead to anergy. In genetic vaccines targeting the macrophages and APC for efficient antigen presentation might lead to balanced immune response. One such approach is to incorporate APC specific promoter in the vector to be used. Methods Three promoters known to be active in macrophage were selected and cloned in mammalian expressing vector pAcGFP1-N1 to reconstruct pAcGFP-MS pAcGFP-EMR and pAcGFP-B5I with macrosialin EmrI and Beta-5 Integrin promoters respectively. As a positive control pAcGFP-CMV was used with CMV promoter and promoterless vector pAcGFP-NIX which served as a negative control. GFP gene was used as readout under the control of each of the promoter. The expression of GFP was analyzed on macrophage and non-macrophage cell lines using Flow cytometry and qRT-PCR with TaqMan probe chemistries. Results All the promoters in question were dominant to macrophage lineage cell lines as observed by fluorescence Western blot and quantitative RT-PCR. The activity of macrosialin was significantly higher than other macrophage promoters. CMV promoter showed times higher activity in macrophage cell lines. The expression of GFP driven by macrosialin promoter after 24 hours was times higher in macrophage derived cell lines in comparison with non macrophage cell lines. Conclusions Based on this study macrosialin promoter can be utilized for targeting macrophage dominant expression. In vivo study needs to be carried out for its utility as a vaccine candidate. Background DNA vaccination wherein plasmid DNA encoding the desired antigen is inoculated in the host is thought to be one of the best approaches to .

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