GFP-PMEL CD8+ T cells were isolated from the spleen of GFP-PMEL TCR transgenic mice through negative selection on microbeads (Miltenyi Biotech, Auburn, CA) according to the manufacturer’s instructions. Purified CD8+ T cells (1 × 106) were injected i.v. into sublethally irradiated (500 rads one day prior to transfer) Krt14-Kitl* hosts (12–16 wk of age). Recipient mice also received i.p. injection of 1 × 106 pfu rVV-hPMEL (N. Restifo, NCI, NIH) on the same day of transfer. IFN-γ blockade was through i.p. injection of 100–500 μg of IFN-γ neutralizing antibody (XMG-6) beginning at the indicated times and twice weekly for the duration of the observation period (5–7 wks). Mice used as controls for vitiligo were sublethally irradiated but did not receive rVV-hPMEL or PMEL CD8+ T cell transfer. Mice with vitiligo used as treatment controls for IFN-γ blockade were treated with either no antibody or with an equal quantity of Rat IgG.
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Adoptive Transfer of GFP-PMEL CD8+ T Cells
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Corresponding Organization :
Other organizations : University of Massachusetts Chan Medical School, University of Pennsylvania, Centenary Institute, University of Sydney, Beth Israel Deaconess Medical Center, Harvard University
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Variable analysis
independent variables
- Administration of IFN-γ neutralizing antibody (XMG-6)
- Development of vitiligo
- Isolation of GFP-PMEL CD8+ T cells from the spleen of GFP-PMEL TCR transgenic mice
- Injection of purified CD8+ T cells (1 × 10^6) i.v. into sublethally irradiated (500 rads one day prior to transfer) Krt14-Kitl* hosts (12–16 wk of age)
- Injection of 1 × 10^6 pfu rVV-hPMEL i.p. into recipient mice on the same day of T cell transfer
- Mice with vitiligo used as treatment controls for IFN-γ blockade were treated with either no antibody or with an equal quantity of Rat IgG
- Mice used as controls for vitiligo were sublethally irradiated but did not receive rVV-hPMEL or PMEL CD8+ T cell transfer
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