Cd4 cd25 isolation kits

We adoptively transferred 10⁶ Teffs, 10⁶ CD4s and either no Tregs or 10⁵ Tregs (WT, CXCR3^−/− or CCR6^−/− Tregs) into 8-12-week-old Rag^−/− hosts infected i.p. with 10⁶ PFU of rVV-hPMEL on the same day of adoptive transfer. Hosts were irradiated with 500 rads 1 day before transfer. In competitive Rag^−/− vitiligo models, vitiligo was induced by adoptive transfer of 10⁶ Thy1.1+ Teffs, 10⁶ CD45.1+ CD4+ T cells, and 5.0×10⁴ Thy1.1+Thy1.2+ WT and 5.0×10⁴ Thy1.2+ KO Tregs (CXCR3^−/− or CCR6^−/−) into 8-12-week-old Rag^−/− hosts irradiated with 500 rads 1 day before transfer, and hosts were infected i.p. with 10⁶ PFU of rVV-hPMEL on the same day of adoptive transfer. Teffs were isolated using a Miltenyi Biotec CD8 isolation kits. Treg-depleted CD4+ T cells were isolated from the spleens of C57BL/6J or CD45.1 mice through negative selection using Miltenyi Biotec CD4+CD25+ isolation kits (Bergisch Gladbach, Germany). Tregs were isolated from the spleens of C57BL/6J mice, CXCR3^−/− mice, or CCR6^−/− mice through positive selection using Miltenyi Biotec CD4+CD25+ isolation kits.