Optimizing the Testing of VLR-CAR in Lymphoid Cells Open Access
Querrey, Melissa Lee (2016)
Abstract
Gene therapy has not only become an increasingly popular method for treating and curing genetic disorders but also in engineering cells for cancer treatment. Through utilization of chimeric antigen receptors (CAR), which include variable lymphocyte receptors (VLR), immunocompetent cells can be directed to target specific antigens common to malignant tumors. To aid in the advancement of this technology, we sought to optimize the method used to efficiently test various VLR containing CAR constructs by looking at gene transfer methods involving transient transfection methods and viral transduction using both AAV and lentivirus-based vectors into lymphoid cells. We have determined an efficient method to rapidly test expression and function of several VLR structures through cells via lentiviral transduction. The ability to use a high-throughput screening method to test for functional VLR-CAR constructs will enhance the process for VLR-CAR characterization and possible selection for clinical translation. However, whether this method is a high-throughput system is yet to be determined. Due to low transduction efficiency of VLR-CAR constructs into more clinically relevant immunocompetent cells with lentivirus, current efforts are focused on an in vivo bone marrow transplant model to proceed testing of the functionality of the VLR-CAR constructs. This study is a gateway to the development of alternative cancer immunotherapies with immunocompetent cells.
Table of Contents
Introduction...1
Methods...10
Results...20
Discussion...27
References...34
Tables, Graphs, and Figures Figure 1...40
Figure 2...41
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Table 3...56
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Optimizing the Testing of VLR-CAR in Lymphoid Cells () | 2018-08-28 13:48:47 -0400 |
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