The profiles of T cells in the ovaries. (A-B) CD45.1⁺ B6 splenocytes (8 × 10⁷) were transplanted into unirradiated allogeneic B6D2F1 (CD45.2⁺) or congenic B6 (CD45.2⁺) mice on day 0. Ovaries are harvested on day +14 after SCT for flow cytometric analysis. (A) Panels show the gating strategies of donor CD4⁺ and CD8⁺ T cells, and the histogram shows detection of donor cells in allogeneic (red line) and syngeneic (blue shaded) hosts. (B) Numbers of donor T cells in the ovaries in syngeneic (n = 6) and allogeneic (n = 10) mice were shown as mean ± SE. (C) Mice were injected with either 3 × 10⁷ CD4⁺ or CD8⁺ T cells from EGFP⁺ mice. Double-label immunofluorescent staining with cleaved-caspase 3 (green) and EGFP (red) of the ovary on day +14. Original magnification ×40. Scale bar, 50 µm. (D-G) Isolated granulosa cells (2 × 10⁴) from the B6D2F1 ovaries were cultured alone (n = 3) or in the presence of 5 × 10⁵ donor T cells sorted from syngeneic (n = 3) or allogeneic recipients (n = 4) on day +15 for 12 hours. Max, maximum. Representative histograms (D) and mean fluorescent intensity (MFI) of granzyme B expression (E) in syngeneic (blue line and bar) or allogeneic (red line and bar) CD8⁺ T cells and isotype controls (shaded area in histogram). Representative FACS plot of PI and Annexin V labeling in CD4⁻CD8⁻ granulosa cells (F) and proportion of AnnexinV⁻ PI⁻ in granulosa cells (G) are shown as mean ± SE. Data from a representative of 2 similar experiments are shown. (H) Transcriptome of donor T cells isolated from syngeneic spleens (n = 2-3), allogeneic spleens (n = 2-3), and allogeneic ovarian follicles (n = 2-4) on day +14 was analyzed. Results of a hierarchical clustering analysis on differentially expressed genes are shown. *P < .05, ***P < .005.