Evidence for Contribution of CD4+CD25+ Regulatory T Cells in Maintaining Immune Tolerance to Human Factor IX following Perinatal Adenovirus Vector Delivery

Figure 1

Measurement of concentrations of hFIX and anti-hFIX antibodies following perinatal and adult AdhFIX injection. (a) Concentrations of hFIX (red upper graph) and anti-hFIX antibodies (black lower graph) following fetal (, small squares) and neonatal (, large diamonds) injection of AdhFIX. (b) Concentrations of hFIX (red upper graph) and anti-hFIX antibodies (black lower graph) following adult injection of AdhFIX. (c) Concentrations of hFIX (red upper graph) and anti-hFIX antibodies (black lower graph) in mice injected in utero with AdhFIX. Two immune challenges with hFIX plus adjuvant and one AdhFIX were administered at , 175, and 275 (labelled arrows). (d) Concentration of anti-adenovirus antibody in naive controls (C1-3), positive control mice which received hFIX plus adjuvant (S1-3), and mice having received AdhFIX in utero (plotted lines). Mice having received AdhFIX in utero received immune challenges with hFIX plus adjuvant and one AdhFIX was administered at , 175, and 275 (labelled arrows). (e) Anti-hFIX antibodies in serially diluted plasma of mice tolerized to hFIX by fetal AdhFIX. Mice received three injections of anti-CD25+ depleting antibody (black, closed circles, and unbroken line) or rat IgG negative control (blue, open circles, and dotted line) simultaneously with hFIX plus ovalbumin plus adjuvant. Nine days after the final challenge, sera were collected for analysis. (f) Anti-ovalbumin antibodies in serially diluted plasma of mice tolerized to hFIX by fetal AdhFIX. Mice received three injections of anti-CD25+ depleting antibody (black, closed circles, and unbroken line) or rat IgG negative control (blue, open circles, and dotted line) simultaneously with hFIX plus ovalbumin plus adjuvant. Nine days after the final challenge, sera were collected for analysis.

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