Separating Lentiviral Vector Injection and Induction of Gene Expression in Time, Does Not Prevent an Immune Response to rtTA in Rats
Open accessAbstract
BACKGROUND: Lentiviral gene transfer can provide long-term expression of therapeutic genes such as erythropoietin. Because overexpression of erythropoietin can be toxic, regulated expression is needed. Doxycycline inducible vectors can regulate expression of therapeutic transgenes efficiently. However, because they express an immunogenic transactivator (rtTA), their utility for gene therapy is limited. In addition to immunogenic proteins that are expressed from inducible vectors, injection of the vector itself is likely to elicit an immune response because viral capsid proteins will induce "danger signals" that trigger an innate response and recruit inflammatory cells. METHODOLOGY AND PRINCIPAL FINDINGS: We have developed an autoregulatory lentiviral vector in which basal expression of rtTA is very low. This enabled us to temporally separate the injection of virus and the expression of the therapeutic gene and rtTA. Wistar rats were injected with an autoregulatory rat erythropoietin expression vector. Two or six weeks after injection, erythropoietin expression was induced by doxycycline. This resulted in an increase of the hematocrit, irrespective of the timing of the induction. However, most rats only responded once to doxycycline administration. Antibodies against rtTA were detected in the early and late induction groups. CONCLUSIONS: Our results suggest that, even when viral vector capsid proteins have disappeared, expression of foreign proteins in muscle will lead to an immune response.
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In vivo application of the Tet-On systems can be complicated by immune responses against rtTA, as reported in rodents and non-human primates.
“In vivo application of the systems can be complicated by immune responses against rtTA, as reported in rodents and non-human primates [73-78].”
From Tet-On Systems For Doxycycline-inducible Gene Expression (2016) · cites this paper as [77]
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- Authors
- Markusic, D. M., de Waart, D. R., & Seppen, J.
- Affiliations
- Amsterdam UMC Location University of Amsterdam (NL)
- Corresponding author
- Seppen, Jurgen
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2010-04-01
- Volume
- 5
- Issue
- 4
- Pages
- e9974
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC2848614
- OpenAlex
- W2074510535
- Cited by
- 22
- References
- 31
- Citations by year
- 2026: 1 · 2023: 1 · 2022: 1 · 2020: 1 · 2019: 2 · 2018: 2 · 2016: 1 · 2015: 1 · 2014: 3 · 2013: 3 · 2012: 4
- FWCI
- 1.62
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- UvA-DARE (University of Amsterdam) (publishedVersion, other-oa)
- Data Archiving and Networked Services (DANS) (submittedVersion, other-oa)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- UvA-DARE (University of Amsterdam) (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Pure Amsterdam UMC (submittedVersion)
- Funders
- ZonMw
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (016.026.012)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- lentiviral vectors, doxycycline-inducible expression, erythropoietin, capsid protein, innate immune response, intramuscular injection, antibody response, hematocrit, gene therapy
- MeSH terms
- Animals, Doxycycline, Erythropoietin, Genetic Vectors, Immunity, Time Factors, Viral Proteins, Genetic Therapy, Transcriptional Activation, Trans-Activators, Lentivirus, Rats, Wistar, Rats
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