Imaging System to Monitor Efficacy of Adenovirus-Based Virotherapy Agents

摘要

Our preliminary data establish a number of important key points. Foremost, these results show that adenovirus can be genetically labeled with a fluorescent structural fusion protein through a complete replacement with IX- EGFP in a chimeric context. At least for our pIX-EGFP strategy, the label was incorporated into virions conferring a fluorescent property that allowed detection of individual particles. Ad-IX-EGFP binding and infection could both be detected via the fluorescent label. This capsid-labeling system is applicable to CRAds because it slightly decreased progeny yield but did not affect the cytopathic effect and spread of the virus. Notably, the level of pIX- EGFP fluorescence directly correlated with the amount of progeny production due to its dependence on E1 activity for expression. The data with pIX-EGFP fulfills all the requirements of the ideal monitoring system for CRAds except noninvasive detection which we propose to accomplish. Both our proposed capsid- labeling approaches demonstrate great promise for detection of viral replication and spread and hence monitoring of CRAds.