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. 2015;2015:397879.
doi: 10.1155/2015/397879. Epub 2015 Jan 31.

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

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Free PMC article

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

Megha S Nivsarkar et al. J Immunol Res. .
Free PMC article

Abstract

Following fetal or neonatal gene transfer in mice and other species immune tolerance of the transgenic protein is frequently observed; however the underlying mechanisms remain largely undefined. In this study fetal and neonatal BALB/c mice received adenovirus vector to deliver human factor IX (hFIX) cDNA. The long-term tolerance of hFIX was robust in the face of immune challenge with hFIX protein and adjuvant but was eliminated by simultaneous administration of anti-CD25+ antibody. Naive irradiated BALB/c mice which had received lymphocytes from donors immunised with hFIX developed anti-hFIX antibodies upon immune challenge. Cotransplantation with CD4+CD25+ cells isolated from neonatally tolerized donors decreased the antibody response. In contrast, cotransplantation with CD4+CD25- cells isolated from the same donors increased the antibody response. These data provide evidence that immune tolerance following perinatal gene transfer is maintained by a CD4+CD25+ regulatory population.

Figures

Figure 1
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 (n = 20, small squares) and neonatal (n = 21, 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 t = 140, 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 t = 140, 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.
Figure 2
Figure 2
Adoptive transfer of regulatory cell populations, tissue distribution of vector, and transgene expression. (a) Anti-hFIX antibody concentrations in serum diluted 1/100 following adoptive transfer of lymphocytes from hFIX-sensitised mice (black bar), cotransplantation of sensitised cells plus CD4+CD25+ cells from neonatally tolerized mice (blue bar) or plus CD4+CD25− cells from neonatally tolerized mice (red bar), or adoptive transfer of lymphocytes from naive donors (grey bars). Recipients underwent sublethal irradiation, adoptive transfer, and hFIX-immunisation 24 hours later and blood collection on days 7, 17, and 24. (b) Quantitative PCR analysis of vector genome distribution and (c) luciferase expression six weeks after neonatal injection of AdLuc.

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