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. Author manuscript; available in PMC: 2012 Jul 1.

Published in final edited form as: Biol Blood Marrow Transplant. 2011 Mar 27;17(7):962–969. doi: 10.1016/j.bbmt.2011.01.018

Fig. 1 — Lethally irradiated (B6.Ly5.1 × bm12)F1 (A) or BALB/c (B) mice were transplanted with TCD-BM alone or plus purified CD4⁺ cells at 1 × 10⁶/mouse from WT or ICOS^-/- B6 donors. L6-Ig or CTLA4-Ig was injected i.p. at 100 μg/mouse every other day for a total of eight doses. Data were obtained for one experiment in each model, and 5-6 mice were included in each group. (C) BMT was set up as in panel A, recipient spleen was collected 6 days after transplantation. Splenocytes were stained individually for surface expression of FasL, intercellular expression of IFNγ or TNFα, in combination with surface expression of CD4 and Ly5.1. The expression of surface FasL (MFI) and intracellular IFNγ or TNFα (% positive) are shown on gated CD4⁺/Ly5.1^- donor cells. The thin lines represent cells stained with isotype control mAb, while thick lines with specific mAb for FasL. The results represent two replicate experiments. (D) BMT was set up as in panel A, peripheral blood was collected from each recipient on day 14. The levels of TNFα, IFNγ, IL-5, IL-2 and IL-4 in the recipient serum were measured as described in Materials and Methods. The levels of IL-2 and IL-4 were under detection level (not shown). The data were pooled from two replicate experiments, and each data point represents a cytokine concentration in one individual mouse.

Fig. 1 — Lethally irradiated (B6.Ly5.1 × bm12)F1 (A) or BALB/c (B) mice were transplanted with TCD-BM alone or plus purified CD4⁺ cells at 1 × 10⁶/mouse from WT or ICOS^-/- B6 donors. L6-Ig or CTLA4-Ig was injected i.p. at 100 μg/mouse every other day for a total of eight doses. Data were obtained for one experiment in each model, and 5-6 mice were included in each group. (C) BMT was set up as in panel A, recipient spleen was collected 6 days after transplantation. Splenocytes were stained individually for surface expression of FasL, intercellular expression of IFNγ or TNFα, in combination with surface expression of CD4 and Ly5.1. The expression of surface FasL (MFI) and intracellular IFNγ or TNFα (% positive) are shown on gated CD4⁺/Ly5.1^- donor cells. The thin lines represent cells stained with isotype control mAb, while thick lines with specific mAb for FasL. The results represent two replicate experiments. (D) BMT was set up as in panel A, peripheral blood was collected from each recipient on day 14. The levels of TNFα, IFNγ, IL-5, IL-2 and IL-4 in the recipient serum were measured as described in Materials and Methods. The levels of IL-2 and IL-4 were under detection level (not shown). The data were pooled from two replicate experiments, and each data point represents a cytokine concentration in one individual mouse.