WT1 specific T cells accumulate in the BM without impairing stem cell function. (A) Donor T cells (CD45.1⁻) were analyzed in the spleen, LNs, and BM of A2Kb Tg mice transplanted with BM stem cells transduced with the WT1-TCR (n = 8) or the LMP2-TCR (n = 5) at 12 weeks after transplantation. The percentage of CD45.1⁻ donor T cells expressing Vβ2.1 (WT1-TCR) or Vβ13 (LMP2-TCR) was determined by FACS analysis. (B) Hematopoietic engraftment in secondary C57Bl/6 recipients (CD45.2) after transplantation of BM cells from A2Kb Tg mice that were previously transplanted with A2Kb BM stem cells (CD45.1) transduced with the WT1-TCR (n = 10). Peripheral blood of secondary recipients was stained with antimurine CD45.1, CD3, B220, and CD11b to identify donor hematopoietic cells, donor T cells, donor B cells and donor granulocytes, respectively. The peripheral blood analysis was done at weeks 3, 4, 8, and 12 after second transplantation. Control C57Bl/6 recipients received BM stem cells from untreated A2Kb (CD45.1) Tg mice (n = 5). Percentage of donor-derived cells are shown after gating on total granulocytes, B cells, and T cells, respectively. *P < .05, **P < .01, ***P < .001. ns indicates not significant.