Adoptive Transfer of Specific CD8+ T Cells

Based on a previously reported protocol (14 (link)), a mixture of 1.5 µg mouse IL-4 (Peprotech, Princeton, NJ, USA) and 50 µg anti-IL-4 antibody (11B11; Bio X Cell, West Lebanon, NH, USA) was intraperitoneally injected into mice daily. After 7 days of treatment, lymphoid organs were analyzed. For the adoptive transfer, donor CD8 T cells were isolated from CD45.1 ⁺ B6 mouse splenocytes using anti-CD8 microbeads and magnetic sorting (MACS; Miltenyi Biotec, Auburn, CA, USA). Next, these CD8 T cells were sorted into CD44^lowCXCR3 ^- and CD44^hiCXCR3 ⁺ populations using a FACSAria (BD Bioscience, San Jose, CA, USA) and then labeled with Cell Trace Violet (CTV, Life Technologies, Waltham, MA, USA) for tracing. Each recipient B6 mouse received 3×10⁶ sorted cells via lateral tail vein injection, followed by IL-4C treatment one day later.