FIG 5.

Enhanced CD11b⁺ Gr1⁺ cell phenotype of CD11c/Nef Tg mice can be observed in recipient mice transplanted with Tg fetal liver (FL) cells. Lethally irradiated non-Tg CD45.2⁺ mice were transplanted with FL cells from CD45.1⁺ CD45.2⁺ Tg or non-Tg E14.5 embryos (A to H) or with mixed (1:1) CD45.2⁺/CD45.1⁺ Tg and non-Tg or control CD45.1⁺ non-Tg FL cells (I). Recipient mice were analyzed 3 to 5 months after transplantation. (A and B) Representative FACS profiles (A) and percentage of donor CD45.1⁺ CD11b⁺ Gr1⁺ cells (B) in spleen, blood, and BM of recipient mice (n = 7 Tg and 5 non-Tg). (C and D) Nonmixed (100%) FL cell transplanted mice. (C) Percentages are of donor spleen CD45.1⁺ CD45.2⁺ CD11b⁺ Gr1⁺ (Gr1^low and Gr1^hi) cells expressing F4/80, MHC class I, MHC class II, CD80, and CD86 in recipient mice (n = 6 to 12) after FL cell transplantation. (D) Mean fluorescence intensity (MFI) of the indicated markers in donor CD45.1⁺ CD11b⁺ Gr1^low and in CD11b⁺ Gr1^hi cell subsets of recipient mice is shown. (E) Proliferation of sorted-purified donor CD11b⁺ Gr1⁺ spleen cells from non-Tg (n = 4) and Tg (n = 5) mice assessed by BrdU labeling. (F and G) Survival of donor CD45.1⁺ CD11b⁺ Gr1⁺ spleen cells ex vivo (F) and after their purification and cultured in vitro in the presence or not of LPS (1 μg/ml) for 24 h (G) was assessed by FACS with 7-AAD labeling. (H and I) Donor (CD45.1⁺ CD45.2⁺) BM progenitors from nonmixed chimera (n = 6 non-Tg and 8 Tg) (H) or donor (CD45.1⁺ and CD45-1⁺ CD45-2⁺) CD11b⁺ Gr1⁺ cells in spleen of mixed chimeric mice (n = 6 non-Tg and 6 Tg chimera) (I) were analyzed by FACS and percentages calculated as for Fig. 4A. NS, not significant. Results are from 3 independent experiments for all phenotypes. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.