A Comparative Study on Delivery of Externally Attached DNA by Papillomavirus VLPs and Pseudoviruses
Abstract
1. Introduction
2. Material and Methods
2.1. L1 VLPs, L1/L2 VLPs, and PSV Production and Validation
2.2. Cells for Externally Attached DNA Delivery
2.3. External DNA Delivery Using VLPs and PSVs in In Vitro Cell Cultures
2.4. Fluorescent Detection and Quantification of Delivery Efficacy Using Flow Cytometry Analysis
2.5. VLP Delivery of Viral DNA for In Vivo Infection of Rabbits
2.6. Statistics
3. Results
3.1. L2 Is Required for Delivering Externally Attached DNA to the Cells In Vitro
3.2. PSVs and VLPs Deliver Attached DNA to Both Rabbit and Human Cells
3.3. PSVs Were More Efficient in Delivering DNA When Encapisidating a Papillomavirus Genome DNA
4. Optimal VLP/External DNA Ratio for Delivery
4.1. PSVs Deliver DNA More Efficiently in Immortalized Cells vs. Normal Cells
4.2. L2 Played a Critical Role in Delivery Efficiency
4.3. VLP-CRPV Increased Viral Infectivity In Vivo
5. Discussion
6. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Name | L1 | L2 | DNA Plasmid | Confirmation by ELISA | Notes |
---|---|---|---|---|---|
L1 VLPs | Yes | No | No | Antibodies used for L1: H16.V5 for HPV16L1, H58.G5 for H58L1, and CRPV.A4 for CRPV L1 L2: (R972 1:100) | All VLPs and PSVs were produced in 293TT cell culture, purified using OptiPrep gradient ultracentrifugation, and stored at −20 °C. For VLPs and PSVs, the positive L1 and L2 fractions were pooled, and concentrations were determined by Bio-Rad protein assay |
VLPs (L1/L2) | Yes | Yes | No | ||
PSVs | Yes | Yes | Yes, (pSEAP, 5.5 kb, PCX 5.5 kb, and the CRPV genome 7.8 kb) |
Dose (ng) | Tumor Outgrowth (Tumor Sites/Challenge Sites) | Average Tumor Size (Geo Mean in mm) | |
---|---|---|---|
DNA | VLPs | ||
10 | - | 1/4 (25%) | 0.5000 |
10 | 20 | 1/2 (50%) | 3.0350 |
20 | - | 2/4 (50%) | 3.8675 |
20 | 40 | 2/2 (100%) | 7.8550 |
40 | - | 3/4 (75%) | 6.3750 |
40 | 80 | 2/2 (100%) | 7.2500 |
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Brendle, S.; Cladel, N.; Balogh, K.; Alam, S.; Christensen, N.; Meyers, C.; Hu, J. A Comparative Study on Delivery of Externally Attached DNA by Papillomavirus VLPs and Pseudoviruses. Vaccines 2021, 9, 1501. https://doi.org/10.3390/vaccines9121501
Brendle S, Cladel N, Balogh K, Alam S, Christensen N, Meyers C, Hu J. A Comparative Study on Delivery of Externally Attached DNA by Papillomavirus VLPs and Pseudoviruses. Vaccines. 2021; 9(12):1501. https://doi.org/10.3390/vaccines9121501
Chicago/Turabian StyleBrendle, Sarah, Nancy Cladel, Karla Balogh, Samina Alam, Neil Christensen, Craig Meyers, and Jiafen Hu. 2021. "A Comparative Study on Delivery of Externally Attached DNA by Papillomavirus VLPs and Pseudoviruses" Vaccines 9, no. 12: 1501. https://doi.org/10.3390/vaccines9121501
APA StyleBrendle, S., Cladel, N., Balogh, K., Alam, S., Christensen, N., Meyers, C., & Hu, J. (2021). A Comparative Study on Delivery of Externally Attached DNA by Papillomavirus VLPs and Pseudoviruses. Vaccines, 9(12), 1501. https://doi.org/10.3390/vaccines9121501