1. Volume 8, Issue 2, Pages 274-283 (August 2003)
Identification of an HLA-A*0201-restricted epitopic peptide from human dystrophin: application in duchenne muscular dystrophy gene therapy 
F Ginhoux, C Doucet, M Leboeuf, F.A Lemonnier, O Danos, J Davoust, H Firat 
Molecular Therapy 
Volume 8, Issue 2, Pages (August 2003)
DOI: /S (03)
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

2. FIG. 1
Human dystrophin expression after human dystrophin DNA transfer in mice. Muscles were harvested at 22 days after DNA injection, quickly frozen in isopentane, and sectioned. Transverse serial sections of tibialis anterior muscle from HHD mice injected with human DYST pTg11025 were immunostained for human dystrophin (A). Positive fibers are presented with at 3× original magnification (B). Micrographs are representative of four experiments. Scale bar, 30 μm.
Molecular Therapy 2003 8, DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

3. FIG. 2
HLA-A*0201-restricted immune responses after human DYST DNA transfer in mice. Spleen and draining lymph nodes were harvested 22 days after DNA injection. (A) For IFN-γ ELISpot assay, splenocytes were cultured in the presence of each human DYST (hDyst) and mouse DYST (mDyst) candidate peptide, for 24 h. Peptide-specific spot numbers are represented after subtraction of the background (2 to 5 spots for irrelevant peptide). (B) For CTL 51Cr-release assay against RMA-S/HHD cells pulsed with human DYST or control influenza matrix I9V peptide, the measurements were performed after 6 days of in vitro stimulation using spleen cells from pTg11025-injected HHD mice or draining lymph node cells (same results, not shown). (C) For the detection of tetramer-positive antigen-specific CD8+ T lymphocytes after two rounds of in vitro stimulation, stimulated lymph node T cells were analyzed using control MelanA/Mart-1 HLA-A*0201 (left) or the human DYST1281 HLA-A*0201 (right) tetramers. The percentage of tetramer-positive CD8+ cells relative to the total CD8+ population is indicated in the upper right quadrant. Results are representative of two to four experiments for each graph.
Molecular Therapy 2003 8, DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions

4. FIG. 3
Flow cytometry analysis of the frequency of human DYST1281 tetramer-positive CD8+ T lymphocytes in secondary lymphoid organs and blood. (A) Spleen cells from HHD control uninjected mice (left) or mice injected 4 weeks and 1 week prior to analysis with human DYST1281 peptide (hDyst1281, middle) or with human DYST DNA (right) as labeled with CD8β and HLA-A*0201-human DYST1281 tetramers are represented. (B) Same staining as in A, using lymph node cells and (C) peripheral blood cells (C) from HHD mice injected with human DYST DNA. Dot plots show the anti-CD8β+ versus human DYST1281-specific (B, right, and C, middle and right) or MelanA/Mart-1 nonspecific HLA tetramer staining (B and C, left). The percentage of tetramer-positive CD8+ cells relative to the total CD8+ population is indicated in the upper right quadrants (A, B, and C). Results are representative of two to four experiments per condition.
Molecular Therapy 2003 8, DOI: ( /S (03) )
Copyright © 2003 The American Society of Gene Therapy Terms and Conditions