Figure 5. RTrx1 affects T cell response to alloantigen in vivo.

Purified T cells from B6 mice were injected i.v. into lethally irradiated BALB/c mice at 0.5 × 10⁶ per mouse. One group of recipient mice were injected i.p. with human RTrx1 at 5 μg/mouse/day from day –1 to day 14. Recipient spleens and liver were collected 21 days after transplant and subjected to cell counting and FACS staining. (A) CD4 and CD8 expression is shown on donor-derived (H2K^b+) live spleen or liver cells, respectively. (B–D) The MFI of DCF-DA is shown on gated donor CD4⁺ and CD8⁺ cells (B), with quantified data shown in C and D. Purified T cells from B6 mice were labeled with CFSE and injected i.v. into lethally irradiated BALB/c mice at 2 × 10⁶ per mouse. One group of recipient mice was injected i.p. with recombinant human RTrx1 at 5 μg/mouse/day from day –1 to day 3. Four days after cell transfer, spleens were collected from recipient mice and subjected to cell counting and FACS staining. (E) CD4 and CD8 expression is shown on donor-derived (H2K^b+) live splenocytes. (F and G) CFSE dilution and percentage of IFN-γ⁺ cells are shown on gated donor CD4⁺ or CD8⁺ cells. (H and I) The mean ± SD is depicted for 4–5 mice per group. Data shown are from 1 representative experiment with mean ± SD of 5 mice per group. Two replicate experiments were performed with a total of 8 mice. Significance was determined by Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001.