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. 2010 Apr 27;18(7):1318–1329. doi: 10.1038/mt.2010.73

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Copyright © 2010 The American Society of Gene & Cell Therapy

This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

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Expansion of CD4⁺IL-10⁺FoxP3⁺ T-cells in response to canine FIX (cFIX) antigens in dogs receiving an AAV-2-cFIX vector via ATVRX. (a) Top: ELISpot assay for IL-10 secretion after stimulation with purified canine FIX protein. Bottom: western blot analysis of cFIX protein in liver lysates from normal and HB dogs. (b) Staining for CD4⁺IL-10⁺FoxP3⁺ T-cells after in vitro restimulation with either cFIX protein (red line), peptide 68 from cFIX (blue line), or cFIX irrelevant pool (gray shaded area). Left panels, histogram plots showing IL-10 intracellular staining on CD4⁺ T-cells; right panels, FoxP3 staining on cells gated on CD4⁺IL-10⁺ T-cells. Cells were gated on lymphocytes, CD4⁺CD8⁻. ATVRX, afferent transvenular retrograde extravasation; AAV, adeno-associated virus; ELISpot, enzyme-linked immunosorbent spot; HB, hemophilia B; IL, interleukin.