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Methods Mol Biol. 2011;807:259-72. doi: 10.1007/978-1-61779-370-7_11.

Measuring immune responses to recombinant AAV gene transfer.

Author information

1
Department of Pharmaceutical Sciences, St. John's University, Queens, NY, USA.

Abstract

Following AAV-based gene transfer, the occurrence of adaptive immune responses specific to the vector or the transgene product is a major roadblock to successful clinical translation. These responses include antibodies against the AAV capsid, which can be neutralizing and therefore prevent the ability to repeatedly administer the vector, and CD8(+) cytotoxic T lymphocytes, which can eliminate transduced cells. In addition, humans may have both humoral and cellular preexisting immunity, as a result from natural infection with parent virus or related serotypes. The need for assays to detect and measure these anti-capsid immune responses in humans and in experimental animals is profound. Here, ELISPOT, immunocapture (ELISA), and neutralization assays are explained and provided in detail. Furthermore, such techniques can readily be adapted to monitor and quantify immune responses against therapeutic transgene products encoded by the vector genome.

PMID:
22034034
PMCID:
PMC3593270
DOI:
10.1007/978-1-61779-370-7_11
[Indexed for MEDLINE]
Free PMC Article

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