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Updated: May 31, 2026
Ex Vivo Infection of Live Tissue with Oncolytic Viruses
Published on: June 25, 2011
Ex vivo infection of live tissue with oncolytic viruses
Jean-Simon Diallo1, Dominic Roy, Hesham Abdelbary
1Center for Innovative Cancer Research, Ottawa Hospital Research Institute (OHRI).
Abstract:
Oncolytic Viruses (OVs) are novel therapeutics that selectively replicate in and kill tumor cells(1). Several clinical trials evaluating the effectiveness of a variety of oncolytic platforms including HSV, Reovirus, and Vaccinia OVs as treatment for cancer are currently underway(2-5). One key characteristic of oncolytic viruses is that they can be genetically modified to express reporter transgenes which makes it possible to visualize the infection of tissues by microscopy or bio-luminescence imaging(6,7). This offers a unique advantage since it is possible to infect tissues from patients ex vivo prior to therapy in order to ascertain the likelihood of successful oncolytic virotherapy(8). To this end, it is critical to appropriately sample tissue to compensate for tissue heterogeneity and assess tissue viability, particularly prior to infection(9). It is also important to follow viral replication using reporter transgenes if expressed by the oncolytic platform as well as by direct titration of tissues following homogenization in order to discriminate between abortive and productive infection. The object of this protocol is to address these issues and herein describes 1. The sampling and preparation of tumor tissue for cell culture 2. The assessment of tissue viability using the metabolic dye alamar blue 3. Ex vivo infection of cultured tissues with vaccinia virus expressing either GFP or firefly luciferase 4. Detection of transgene expression by fluorescence microscopy or using an In Vivo Imaging System (IVIS) 5. Quantification of virus by plaque assay. This comprehensive method presents several advantages including ease of tissue processing, compensation for tissue heterogeneity, control of tissue viability, and discrimination between abortive infection and bone fide viral replication.
Insights
This protocol details a method for ex vivo infection of tumor tissues with oncolytic viruses (OVs). It ensures accurate assessment of viral replication and tissue viability for effective cancer virotherapy.
Area of Science:
- Oncolytic virotherapy
- Cancer research
- Molecular virology
Background:
- Oncolytic viruses (OVs) are emerging cancer therapeutics that selectively target and destroy tumor cells.
- Genetically engineered OVs can express reporter transgenes for visualizing infection and assessing therapeutic potential.
- Ex vivo infection of patient tissues is crucial for predicting treatment success and managing tissue heterogeneity.
Purpose of the Study:
- To describe a comprehensive protocol for ex vivo infection of tumor tissues with OVs.
- To detail methods for tissue preparation, viability assessment, and viral infection.
- To enable accurate evaluation of oncolytic virus replication and efficacy prior to in vivo administration.
Main Methods:
- Tumor tissue sampling and preparation for cell culture.
- Assessment of tissue viability using the alamar blue metabolic dye.
- Ex vivo infection with vaccinia virus (reporter transgenes: GFP or firefly luciferase).
- Detection of transgene expression via fluorescence microscopy or In Vivo Imaging System (IVIS).
- Quantification of viral replication using plaque assays.
Main Results:
- The protocol facilitates easy tissue processing and accounts for tissue heterogeneity.
- It allows for precise control of tissue viability before infection.
- The method effectively distinguishes between abortive and productive viral infections.
Conclusions:
- This protocol provides a robust framework for ex vivo assessment of oncolytic virus efficacy.
- It enhances the predictability of oncolytic virotherapy outcomes.
- The method supports personalized cancer treatment strategies through pre-therapy tissue analysis.
