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Major role of local immune responses in antibody formation to factor IX in AAV gene transfer

Gene Therapy volume 12, pages 1453–1464 (2005)Cite this article

Abstract

The risk of an immune response to the coagulation factor IX (F.IX) transgene product is a concern in gene therapy for the X-linked bleeding disorder hemophilia B. In order to investigate the mechanism of F.IX-specific lymphocyte activation in the context of adeno-associated viral (AAV) gene transfer to skeletal muscle, we injected AAV-2 vector expressing human F.IX (hF.IX) into outbred immune-competent mice. Systemic hF.IX levels were transiently detected in the circulation, but diminished concomitant with activation of CD4+ T and B cells. ELISPOT assays documented robust responses to hF.IX in the draining lymph nodes of injected muscle by day 14. Formation of inhibitory antibodies to hF.IX was observed over a wide range of vector doses, with increased doses causing stronger immune responses. A prolonged inflammatory reaction in muscle started at 1.5–2 months, but ultimately failed to eliminate transgene expression. By 1.5 months, hF.IX antigen re-emerged in circulation in ∼70% of animals injected with high vector dose. Hepatic gene transfer elicited only infrequent and weaker immune responses, with higher vector doses causing a reduction in T-cell responses to hF.IX. In summary, the data document substantial influence of target tissue, local antigen presentation, and antigen levels on lymphocyte responses to F.IX.

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Acknowledgements

This work was supported by NIH Grants R01 AI/HL51390-01 and U01 HL66948 to RWH. ED was supported by NIH training grant T32HL07439. We thank the staff of the PEGT Vector Core at The Children's Hospital of Philadelphia for technical assistance.

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Authors and Affiliations

  1. Department of Pediatrics, University of Pennsylvania Medical Center, Philadelphia, PA, USA

    L Wang, O Cao, B Swalm, E Dobrzynski, F Mingozzi & R W Herzog

  2. The Children's Hospital of Philadelphia, Philadelphia, PA, USA

    L Wang, O Cao, B Swalm, E Dobrzynski, F Mingozzi & R W Herzog

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  1. L Wang
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  2. O Cao
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Wang, L., Cao, O., Swalm, B. et al. Major role of local immune responses in antibody formation to factor IX in AAV gene transfer. Gene Ther 12, 1453–1464 (2005). https://doi.org/10.1038/sj.gt.3302539

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