Fig. 3.

Retroviral transduction of Egr-2 into naïve T cells. (A) Retroviral constructs for the transduction of Egr-2. (B) Ectopic Egr-2 expression induced the expression of LAG-3, IL-10, and Blimp-1. Quantitative PCR analyses of gene expression in sorted retrovirally transduced cell populations stimulated for 5 days with soluble anti-CD3 mAb (1 μg/mL). The results are the means of three independent experiments. (C) Cytokines in the culture supernatants of pMIG- and pMIG-Egr2-transduced CD4⁺GFP⁺ T cells stimulated for 48 h with or without plate-coated anti-CD3 mAb. The results are the means of three independent experiments. (D) Suppression of naïve carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled CD4⁺CD25⁻CD45RB^high T cells by Egr-2-transduced T cells in vitro. Naïve CD4⁺CD25⁻CD45RB^high Thy1.1⁺ T cells were labeled with CFSE and cultured with the indicated retrovirally transduced Thy1.2⁺ T cells and irradiated whole splenocytes plus anti-CD3 mAb. Representative data from three independent experiments are shown. (E) Antigen-specific suppression of the delayed-type hypersensitivity (DTH) response by Egr2-transduced CD4⁺ T cells. Six days after primary immunization, FACS-sorted pMIG- or pMIG-Egr2-transduced CD4⁺ T cells (1 × 10⁶) from BALB/c or DO11.10 mice were transferred adoptively via i.v. injection into BALB/c mice. Two days after adoptive cell transfer, DTH response in the footpad was induced. Footpad thickness was determined 24 h later; n = 6 per group. All error bars represent ±SD. **, P < 0.01.