Subretinal delivery of adeno-associated virus serotype 2 results in minimal immune responses that allow repeat vector administration in immunocompetent mice
| Autor: | Robin R. Ali, Prateek K. Buch, Scott J Robbie, Robert E MacLaren, Yanai Duran, M. Natkunarajah, Kamaljit S. Balaggan, S. E. Barker, James W B Bainbridge, Alexander J. Smith, Cathryn A Broderick |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
cis-trans-Isomerases
Transgene viruses Genetic Vectors Mice Transgenic RPE65 Biology medicine.disease_cause Eye Virus immune response Viral vector Cell Line Injections Mice Immune system Neutralization Tests antibody Drug Discovery Genetics medicine Electroretinography Animals Eye Proteins Molecular Biology Adeno-associated virus subretinal Genetics (clinical) AAV Genetic Therapy Dependovirus eye diseases Mice Inbred C57BL Cis-trans-Isomerases AAV antibody immune response RPE65 subretinal Immunology biology.protein Molecular Medicine Female sense organs Immunocompetence Antibody Carrier Proteins Research Article |
| Zdroj: | The Journal of Gene Medicine Journal of Gene Medicine, 11 (6) pp. 486-497. (2009) |
| Popis: | Background Adeno-associated virus serotype 2 (AAV2) vectors show considerable promise for ocular gene transfer. However, one potential barrier to efficacious long-term therapy is the development of immune responses against the vector or transgene product. Methods We evaluated cellular and humoural responses in mice following both single and repeated subretinal administration of AAV2, and examined their effects on RPE65 and green fluorescent protein transgene expression. Results Following subretinal administration of vector, splenocytes and T-cells from draining lymph nodes showed minimal activation following stimulation by co-culture with AAV2. Neutralizing antibodies (NAbs) were not detected in the ocular fluids of any mice receiving AAV2 or in the serum of mice receiving a lower dose. NAbs were present in the serum of a proportion of mice receiving a higher dose of the vector. Furthermore, no differences in immunoglobulin titre in serum or ocular fluids against RPE65 protein or AAV2 capsid between treated and control mice were detected. Histological examination showed no evidence of retinal toxicity or leukocyte infiltration compared to uninjected eyes. Repeat administration of low-dose AAV.hRPE65.hRPE65 to both eyes of RPE65−/− mice resulted in transgene expression and functional rescue, but re-administration of high-dose AAV2 resulted in boosted NAb titres and variable transgene expression in the second injected eye. Conclusions These data, which were obtained in mice, suggest that, following subretinal injection, immune responses to AAV2 are dose-dependent. Low-dose AAV2 is well tolerated in the eye, with minimal immune responses, and transgene expression after repeat administration of vector is achievable. Higher doses lead to the expression of NAbs that reduce the efficacy of repeated vector administration. Copyright © 2009 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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