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Immune responses to adeno-associated virus type 2 encoding channelrhodopsin-2 in a genetically blind rat model for gene therapy

Gene Therapy volume 18, pages 266–274 (2011)Cite this article

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Abstract

We had previously reported that transduction of the channelrhodopsin-2 (ChR2) gene into retinal ganglion cells restores visual function in genetically blind, dystrophic Royal College of Surgeons (RCS) rats. In this study, we attempted to reveal the safety and influence of exogenous ChR2 gene expression. Adeno-associated virus (AAV) type 2 encoding ChR2 fused to Venus (rAAV-ChR2V) was administered by intra-vitreous injection to dystrophic RCS rats. However, rAAV-ChR2 gene expression was detected in non-target organs (intestine, lung and heart) in some cases. ChR2 function, monitored by recording visually evoked potentials, was stable across the observation period (64 weeks). No change in retinal histology and no inflammatory marker of leucocyte adhesion in the retinal vasculature were observed. Although antibodies to rAAV (0.01–12.21 μg ml−1) and ChR2 (0–4.77 μg ml−1) were detected, their levels were too low for rejection. T-lymphocyte analysis revealed recognition by T cells and a transient inflammation-like immune reaction only until 1 month after the rAAV-ChR2V injection. In conclusion, ChR2, which originates from Chlamydomonas reinhardtii, can be expressed without immunologically harmful reactions in vivo. These findings will help studies of ChR2 gene transfer to restore vision in progressed retinitis pigmentosa.

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Acknowledgements

This work was partly supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (nos. 21791664 and 21200022); Science and Culture and Special Coordination Funds for Promoting Science and Technology of the Japanese Government, Strategic Research Programme for Brain Sciences (SRPBS); the Ministry of Health, Labour and Welfare of Japan; and Japan Foundation for Aging and Health and the Programme for Promotion of Fundamental studies in Health Sciences of the National Institute of Biomedical Innovation (NIBIO). I also thank Teru Hiroi for technical support in animal treatment.

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Authors and Affiliations

  1. Department of International Advanced Interdisciplinary Research, Institute for International Advanced Research and Education, Tohoku University, Sendai, Japan

    E Sugano, H Isago, Z Wang, N Murayama & H Tomita

  2. Japan Foundation for Aging and Health, Aichi, Japan

    Z Wang

  3. School of Medicine, Tohoku University, Sendai, Japan

    M Tamai

  4. United Centers for Advanced Research and Translational Medicine, School of Medicine, Tohoku University, Sendai, Japan

    H Tomita

  5. Innovation of New Biomedical Engineering Center, Tohoku University, Sendai, Japan

    H Tomita

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  1. E Sugano
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Correspondence to H Tomita.

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Sugano, E., Isago, H., Wang, Z. et al. Immune responses to adeno-associated virus type 2 encoding channelrhodopsin-2 in a genetically blind rat model for gene therapy. Gene Ther 18, 266–274 (2011). https://doi.org/10.1038/gt.2010.140

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