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Zhang W.,Guangdong Pharmaceutical University | Zhang W.,Guangdong Province Key Laboratory for Biotechnology Drug Candidate | Shao H.,Guangdong Pharmaceutical University | Shao H.,Guangdong Province Key Laboratory for Biotechnology Drug Candidate | And 4 more authors.
Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | Year: 2014

The research on intracellular trafficking of adenovirus has been described mainly through observations of subgroup C adenoviruses in transformed cell lines. The basic elements of the trafficking pathway include binding to receptors at the cell surface, internalization by endocytosis, analysis of the endosomal membrane, escape to the cytosol, intracellular trafficking along microtubules, nuclear pore docking, and viral genome translocation into the nucleus. More than 80% of the adenovirus genome is delivered to the nucleus in a highly efficient manner in approximately 1 h. However, exceptions to this trafficking pattern have been noted, including: variations based on target cell type, cell physiology, and adenovirus serotype. This review summarizes mechanism of adenovirus infection pathway and intracellular trafficking, providinging a foundation for the development of clinical adenoviral vector. © 2014 Chin J Biotech, All rights reserved.

Shao H.,Guangdong Pharmaceutical University | Shao H.,Guangdong Province Key Laboratory for Biotechnology Drug Candidate | Lin Y.,Guangdong Pharmaceutical University | Lin Y.,Guangdong Province Key Laboratory for Biotechnology Drug Candidate | And 19 more authors.
Cancer Letters | Year: 2015

Identification of TCR genes specific for tumor-associated antigens (TAAs) is necessary for TCR gene modification of T cells, which is applied in anti-tumor adoptive T cell therapy (ACT). The usual identification methods are based on isolating single peptide-responding T cells and cloning the TCR gene by in vitro expansion or by single-cell RT-PCR. However, the long and exacting in vitro culture period and demanding operational requirements restrict the application of these methods. Immunoscope is an effective tool that profiles a repertoire of TCRs and identifies significantly expanded clones through CDR3 length analysis. In this study, a survivin-derived mutant peptide optimized for HLA-A2 binding was selected to load DCs and activate T cells. The monoclonal expansion of TCRA and TCRB genes was separately identified by Immunoscope analysis and following sequence identification, the properly paired TCR genes were transferred into T cells. Peptide recognition and cytotoxicity assays indicated that TCR-modified PBMCs could respond to both the mutant and wild type peptides and lyse target cells. These results show that combining Immunoscope with in vitro peptide stimulation provides an alternative and superior method for identifying specific TCR genes, which represents a significant advance for the application of TCR gene-modified T cells. © 2015 Elsevier Ireland Ltd.

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