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Heparin binding directs activation of T cells against adeno-associated virus serotype 2 capsid

Nature Medicine volume 12, pages 967–971 (2006)Cite this article

Abstract

Activation of T cells to the capsid of adeno-associated virus (AAV) serotype 2 vectors has been implicated in liver toxicity in a recent human gene therapy trial of hemophilia B1. To further investigate this kind of toxicity, we evaluated T-cell responses to AAV capsids after intramuscular injection of vectors into mice and nonhuman primates. High levels of T cells specific to capsids of vectors based on AAV2 and a phylogenetically related AAV variant were detected. Vectors from other AAV clades2 such as AAV8 (ref. 3), however, did not lead to activation of capsid-specific T cells. Through the generation of AAV2-AAV8 hybrids and the creation of site-directed mutations, we mapped the domain that directs the activation of T cells to the RXXR motif on VP3, which was previously shown to confer binding of the virion to heparan sulfate proteoglycan (HSPG)4,5,6. Evaluation of natural and engineered AAV variants showed direct correlations between heparin binding, uptake into human dendritic cells (DCs) and activation of capsid-specific T cells. The role of heparin binding in the activation of CD8+ T cells may be useful in modulating the immunogenicity of antigens and improving the safety profile of existing AAV vectors for gene therapy.

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Figure 1: Activation of T cells in mice after administration of AAV.
Figure 2: Time course of T-cell response to AAV capsid in cynomolgus macaques after intramuscular vaccinations with AAV vectors of different serotypes.
Figure 3: Impact of HSPG affinity on AAV binding.

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Acknowledgements

We would like to acknowledge J. Franco and the support of Z. Abbas, Y. Li, E. McComb, R. Patel, A. Wang and E. Wolf from the Vector Core of the University of Pennsylvania. R. Desai is supported by a graduate research fellowship from the National Science Foundation. Microscopy equipment was financed with grants from the National Heart, Lung, and Blood Institute (HL073305) and National Institute of Biomedical Imaging and BioEngineering (EB00262). The research was funded by grants from GlaxoSmithKline and the US National Institutes of Health (P01-HL-059407 and P30-DK-047757).

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  1. Luk H Vandenberghe and Lili Wang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, Gene Therapy Program, University of Pennsylvania School of Medicine, 125 S. 31st Street, Philadelphia, 19104, Pennsylvania, USA

    Luk H Vandenberghe, Lili Wang, Suryanarayan Somanathan, Yan Zhi, Joanita Figueredo, Roberto Calcedo, Julio Sanmiguel, Julie Johnston, Rebecca L Grant, Guangping Gao & James M Wilson

  2. Molecular Medicine, Katholieke Universiteit Leuven, Kapucijnenvoer 33, Leuven, B-3000, Belgium

    Luk H Vandenberghe

  3. Department of Bioengineering, University of Pennsylvania School of Engineering and Applied Science, 120 Hayden Hall, 3320 Smith Walk, Philadelphia, 19104, Pennsylvania, USA

    Ravi A Desai & Christopher S Chen

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  1. Luk H Vandenberghe
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Correspondence to James M Wilson.

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James M. Wilson is the recipient of a grant from GlaxoSmithKline.

The University of Pennsylvania has licensed some of the vectors described in this paper.

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Vandenberghe, L., Wang, L., Somanathan, S. et al. Heparin binding directs activation of T cells against adeno-associated virus serotype 2 capsid. Nat Med 12, 967–971 (2006). https://doi.org/10.1038/nm1445

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