Bortezomib treatment significantly reduces anti-AAV IgG titre. a.

<p>Mice were infected with AAV2/8-EV and treated with bortezomib, as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034684#s2" target="_blank">Materials and Methods</a> section. Sera were collected at the indicated times, and the levels of anti-AAV IgG were determined by titration. <b>b.</b> A subset of the same sera samples was assayed for the total amount of IgG using ELISA. Representative data from two separate experiments with similar outcomes are shown.</p

Bortezomib-mediated reduction in anti-AAV IgG titres is insufficient to facilitate reinfection with AAV2/8-DC190-alphaGal.

<p><b>a.</b> At twenty-four weeks following the administration of AAV2/8-EV (including 20 weeks of bortezomib treatment between weeks 4 and 24), mice were re-infected with 5×10¹¹ AAV2/8-DC190-alphaGal/mouse vector particles. Sera were collected at the indicated times, and the titres of anti-AAV IgG were determined. <b>b.</b> A subset of the sera samples was assayed for the total amount of IgG using ELISA. <b>c.</b> The level of α-galactosidase A in serum samples from mice infected with AAV2/8-DC190-alphaGal. Expression of α-galactosidase A was observed in mice that had not been previously infected with AAV2/8-EV. <b>d.</b> Immunohistochemistry was performed on liver sections from mice that were administered AAV2/8-DC190-alphaGal. The figures show representative data from two studies with similar outcomes.</p

Bortezomib treatment differentially affects T cells.

<p><b>a.</b> Mononuclear cells were prepared from spleens and analysed for the T cell markers CD4, CD8, and CD44 using flow cytometry. Plots depict CD44 expression gated on the CD4⁺ (left panel) and CD8⁺ (right panel) populations. <b>b.</b> A summary of the data from <b>a</b> is shown in a quantitative format. <b>c.</b> Mononuclear cells isolated from spleen were reactivated with AAV2/8-EV for 72 h <i>in vitro</i>. Supernatants were collected and assayed for IL-2 (left panel) or interferon-γ (right panel) levels using ELISA. The figures show representative data from two studies with similar outcomes.</p