α pe magnetic beads

All mice were used in accordance with the Institutional Animal Care and Use Committee at the University of Minnesota. C57BL/6J mice were purchased from The Jackson Laboratory, P14 and OT-I CD8 T cell transgenic mice were maintained in house. P14 immune chimeras were generated by transferring 5×10⁴ P14 CD8 T cells into naive C57BL/6J mice. The following day these mice were infected with 2×10⁵ PFU LCMV Armstrong via intraperitoneal (i.p) injection. For endogenous studies naive C57BL/6J mice were infected with 2×10⁵ PFU LCMV Armstrong i.p. OT-I immune chimeras were generated by transferring 5×10⁴ naїve OT-I CD8 T cells into C57BL/6 mice. The next day, mice were infected with 2×10⁶ PFU Vaccinia Virus expressing chicken ovalbumin. Sixty days after infection, CD62L+ and CD62L- memory OT-I splenocytes were purified using α-CD62L PE and α-PE magnetic beads according to the manufacturers instructions (Miltenyi). 5×10⁵ CD62L+ or CD62L- OT-I cells were transferred into P14 immune chimeras that 60 days previously had been infected with LCMV. The following day animals were transcervically (t.c.) challenged with 50μg gp-33 peptide as previously described (Collins et al.; Schenkel et al., 2013 (link)). Parabiosis surgeries were performed as previously described (Schenkel et al., 2013 (link)).