Figure 6.

Gr-1 depletion affects CD4⁺CD8⁺ (DP) T cells in N3-tg mice. N3-tg mice were injected i.p. with anti-Gr-1 depleting antibodies (RB6-8C5) or isotype controls (Ctr), twice a week and sacrificed/analyzed after 4 weeks of treatment; some of the mice were also injected i.p with BrdU solution 1 day before the sacrifice. (A) Percentages of CD4⁺CD8⁺ cells are shown, as measured by FACS analysis in the BM of N3-tg mice, treated as above. Numbers inside cytograms indicate the mean percentages ± SD of CD4⁺CD8⁺ cells. (B) The graph reports the ratio between the CD4⁺CD8⁺ cell counts in RB6-8C5-treated N3-tg mice and in relative controls, expressed as % of control, and calculated in the BM and spleen (SPL), by the same FACS analysis, as in (A). In (A, B), the data are presented as the mean value ± SD from three independent experiments (n = 4 mice per group). (C) Representative FACS analysis of BrdU⁺ cells inside the CD4⁺CD8⁺ subset in the BM of N3-tg mice, treated as above, with isotype control antibodies (Ctr) or anti-Gr-1 depleting antibodies (RB6-8C5). Numbers inside cytograms indicate the percentages of CD4⁺CD8⁺BrdU⁺ cells. (D) The graphs show the CD4⁺CD8⁺BrdU⁺ cell percentages in the BM and spleen (SPL) of N3-tg mice, treated as above, and measured by the same FACS analysis, as in (C). We have not observed any effects of anti-Gr-1 treatment on T-cell compartment of wt mice (not shown). (E) The graphs report total cell counts (left panel), absolute numbers of DP T cells (middle panel), and percentages of BrdU⁺DP T cells (right panel), in mice injected with RB6-8C5 antibodies alone (black bars), or in combination with anti-IL-6 antibodies (red bars), compared with controls (white bars). In (D, E), the results are presented as the mean value ± SD from two independent experiments (n = 3 mice per group). ns, not significant, p > 0.05; ^* p ≤ 0.05; ^** p ≤ 0.01, and ^*** p ≤ 0.001 represent significant differences between the indicated groups.