Allogeneic TCD marrow plus highly enriched B6-cdd donor CD4+ T cells induce GVHD-associated weight loss and lethality in BALB/c recipients.
(A) Enrichment of B6 and B6-cdd CD4+ T cells used for BMT. Spleen and lymph node cells were positively selected by means of the Macs system. The enriched T-cell population was stained with anti-CD4 and anti-CD8 mAbs and analyzed with a FACScan. The percentage of contaminating CD8+ cells in each experiment ranged from 0.3% to 1.1%. Comparable results (data not shown) were obtained with the use of Dynal magnetic beads. (B) CD4+ T cells from B6-cdd donors induced weight loss in allogeneic BALB/c BMT recipients. BALB/c recipients (5 per group) irradiated with 8.5 Gy were transplanted with 5 × 106 B6-Ly5.1 TCD–bone marrow cells by means of varying numbers of CD4+ T cells from either wild-type B6 or B6-cdd donors as indicated. Total body weights of recipients were monitored following BMT. Compared with wild-type B6 CD4+ T cells, transplantation of equivalent numbers of B6-cdd CD4+ T cells induced delayed and milder weight loss in BALB/c recipients. (C) CD4+ T cells from B6-cdd donors can affect lethality in BALB/c BMT recipients. The BMT recipient groups described in Figure 2B were examined for survival after transplantation. Recipients of 2.5 × 106 and 1 × 106 B6-cdd CD4+ T cells exhibited mortality. However, there was a significant delay in these groups compared with recipients of wild-type B6 CD4+ T cells (P = .0002). ■, 2.5 × 106 B6-cdd; ▵, 1.0 × 106 B6-cdd; ◊, 2.5 × 106 B6-cdd; ■, 2.5 × 106 B6; ▴, 1.0 × 106 B6-wt; and ⧫, 0.25 × 106 B6 donor CD4+ T cells.
