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Lack of humoral immune response to the tetracycline (Tet) activator in rats injected intracranially with Tet-off rAAV vectors

Gene Therapy volume 17, pages 616–625 (2010)Cite this article

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Abstract

The ability to safely control transgene expression from viral vectors is a long-term goal in the gene therapy field. We have previously reported tight regulation of GFP expression in rat brain using a self-regulating tet-off rAAV vector. The immune responses against tet regulatory elements observed by other groups in nonhuman primates after intramuscular injection of tet-on encoding vectors raise concerns about the clinical value of tet-regulated vectors. However, previous studies have not examined immune responses following injection of AAV vectors into brain. Therefore, rat striatum was injected with tet-off rAAV harboring a therapeutic gene for Parkinson's disease, either hAADC or hGDNF. The expression of each gene was tightly controlled by the tet-off regulatory system. Using an ELISA developed with purified GST–tTA protein, no detectable immunogenicity against tTA was observed in sera of rats that received an intrastriatal injection of either vector. In contrast, sera from rats intradermally injected with an adenovirus containing either tTA or rtTA, as positive controls, had readily detectable antibodies. These observations suggest that tet-off rAAV vectors do not elicit an immune response when injected into rat brain and that these may offer safer vectors for Parkinson's disease than vectors with constitutive expression.

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Acknowledgements

We thank Dr Christina Khodr, Northwestern University, for editorial assistance. Ms Jianping Xie and Mrs Xue Song Wang for their excellent technical support. This work was supported by NIH Grants NS31957 and U54NS045309, the Harry F and Elaine M. Chaddick Foundation and the Medical Research Institute Council of Children's Memorial Hospital. We are also grateful for the support of the Chicago Biomedical Consortium and the State of Illinois Excellence in Academic Medicine Program to the viral vector core at the Children's Memorial Research Center.

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  1. Department of Pediatrics, Neurobiology Program, Children's Memorial Research Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA

    Y Han, Q A Chang, T Virag, N C West, D George & M C Bohn

  2. Cedars-Sinai Medical Center and Department of Molecular and Medical Pharmacology, Gene Therapeutics Research Institute, UCLA, Los Angeles, CA, USA

    M G Castro

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Correspondence to M C Bohn.

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Han, Y., Chang, Q., Virag, T. et al. Lack of humoral immune response to the tetracycline (Tet) activator in rats injected intracranially with Tet-off rAAV vectors. Gene Ther 17, 616–625 (2010). https://doi.org/10.1038/gt.2010.6

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