Figure 3.

Antitransgene CD4⁺ helper T-cell response dictates the magnitude of antitransgene CD8⁺ T-cell responses and early efficacy of miR142-3pT–mediated regulation. (a–b) C57Bl/6 female or male mice were injected i.m. at day 0 with rAAV1-mOVA-HY or rAAV1-mOVA-HY-miR. (a) On day 14 or 35, mice were sacrificed and their splenocytes were tested by IFN-γ ELISPOT assay against OVA₂₅₇ peptide. Data represent 6–10 mice per group and per time point pooled from two to four independent experiments. (b) On day 14, tibialis anterior muscles were frozen and 7 µm thick transversal cryosections were stained with hematoxilin and eosin. Field size is 720 × 720 micron. One representative muscle out of three is shown. (c–d) A group of C57Bl/6 female mice were injected by 60 μg of GK1.5 depleting antibody at day -3, 0, 7, 1, and 21. GK1.5-treated and control C57Bl/6 female mice were injected i.m. at day 0 with rAAV1-mOVA-HY or rAAV1-mOVA-HY-miR. (c) Their PBL were stained to assess the efficiency of CD4 depletion by flow cytometry. Representative dot plots for CD4 and CD8 expression on live PBL at day 14 are shown. (d) On day 14 or 28, mice were sacrificed and their splenocytes tested by IFN-γ ELISPOT assay against OVA₂₅₇ peptide. Data represent 6–10 mice per group and per time point pooled from 2–4 independent experiments. ND, not detected.