Cytotoxic T Lymphocyte Responses to Transgene Product, Not Adeno-Associated Viral Capsid Protein, Limit Transgene Expression in Mice
| Autor: | William M. Siders, Johanne Kaplan, Abraham Scaria, Michael Lukason, Jacqueline Shields, Sam Wadsworth, Lisa Woodworth |
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| Rok vydání: | 2009 |
| Předmět: |
viruses
Genetic enhancement Transgene Genetic Vectors Gene delivery Immunocompromised Host Mice Scientific Papers Genetics Animals Cytotoxic T cell Amino Acid Sequence Transgenes Vector (molecular biology) Neutralizing antibody Molecular Biology Mice Inbred BALB C biology Genetic Therapy Dependovirus Virology Molecular biology Mice Inbred C57BL CTL Capsid alpha-Galactosidase biology.protein Molecular Medicine Capsid Proteins Peptides Immunocompetence T-Lymphocytes Cytotoxic |
| Zdroj: | Human Gene Therapy. 20:11-20 |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/hum.2008.055 |
| Popis: | The use of adeno-associated viral (AAV) vectors for gene replacement therapy is currently being explored in several clinical indications. However, reports have suggested that input capsid proteins from AAV-2 vector particles may result in the stimulation of cytotoxic T lymphocyte (CTL) responses that can result in a loss of transduced cells. To explore the impact of anti-AAV CTLs on AAV-mediated transgene expression, both immunocompetent C57BL=6 mice and B cell-deficient muMT mice were immunized against the AAV2 capsid protein (Cap) and were injected intravenously with an AAV-2 vector encoding alpha-galactosidase (alpha-Gal). C57BL=6 mice, which developed both CTL and neutralizing antibody responses against Cap, failed to show any detectable alpha-Gal expression. In contrast, serum alpha-Gal levels comparable to those of naive mice were observed in muMT mice despite the presence of robust CTL activity against Cap, indicating that preexisting Cap-specific CTLs did not have any effect on the magnitude and duration of transgene expression. The same strategy was used to assess the impact of CTLs against the alpha-Gal transgene product on AAV-mediated gene delivery and persistence of transgene expression. Preimmunization of muMT mice with an Ad=alpha-Gal vector induced a robust CTL response to alpha-Gal. When these mice were injected with AAV2=alpha-Gal vector, initial levels of alpha-Gal expression were reduced by more than 1 log and became undetectable by 2 weeks postinjection. Overall, our results indicate that CTLs against the transgene product as opposed to AAV capsid protein are more likely to interfere with AAV transgene expression. |
| Databáze: | OpenAIRE |
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