Figure 7.
GM-CSF production by donor-derived CD4+ T cells promotes indirect alloantigen presentation. (A-E). Lethally irradiated Balb/c mice were transplanted with B6 Rag-1−/− BM alone (5 × 106) or together with 1 × 106 purified CD4+ T cells from WT or GM-CSF−/− mice. (A) Representative dot plot gated on donor-derived CD45.2+ TCRβ− CD64− mLN cells 13 days posttransplantation depicting coexpression of CD11c and class II (I-Ab). (B) Frequency and absolute number of dendritic cells in the mLN. Results are from 3 experiments with 11 to 13 mice/group. (C) Percentage of dendritic cells in the mLN that coexpressed CD103 and/or CD11b. Data are derived from 2 experiments with 8 to 9 mice/group. (D-E) Representative flow plots and quantification of the expression of host peptide-donor MHC-II complex (YAe) on donor-derived DCs (CD45.2+ TCRβ−, CD64− CD11c+ MHC-II+). Data are from 3 experiments (n = 11-14/group). (F-J) Lethally irradiated Balb/c mice were transplanted with B6 Rag-1−/− BM (5 × 106) together with 1 × 106 purified CD4+ T cells from WT or GM-CSF−/− mice. Control mice received H2-Ab1−/− (MHC-II-deficient) BM (5 × 106) and 1 × 106 purified CD4+ from WT B6 donors. Sort-purified CD45.1+ CD4+ Vα2+ Vβ6+ cells (1 × 106) from TEa mice were adoptively transferred to mice 12 days posttransplantation. (F) Experimental design. (G) Representative dot plot gated on live, singlet cells from the colons of mice receiving H2-Ab1−/− BM and WT CD4+ T cells or Rag-1−/− BM and either WT or GM-CSF−/− CD4+ T cells 7 days posttransfer. (H) The frequency (expressed as a percentage of total live colon lamina propria cells) and absolute number of CD45.1+ CD4+ TEa cells in the colon 7 days posttransfer. Results are from 3 experiments (n = 9-11 per group). (I) Representative flow plots gated on CD4+ CD45.1+ (ie, TEa) cells depicting GM-CSF production. (J) The percentage of CD45.1+ CD4+ TEa cells in the colon of mice that expressed IFN-γ, GM-CSF, or TNF-α 7 days posttransfer. Results are from 2 experiments (n = 9/group). *P < .05; **P < .01; ***P < .001.

GM-CSF production by donor-derived CD4+ T cells promotes indirect alloantigen presentation. (A-E). Lethally irradiated Balb/c mice were transplanted with B6 Rag-1−/− BM alone (5 × 106) or together with 1 × 106 purified CD4+ T cells from WT or GM-CSF−/− mice. (A) Representative dot plot gated on donor-derived CD45.2+ TCRβ− CD64− mLN cells 13 days posttransplantation depicting coexpression of CD11c and class II (I-Ab). (B) Frequency and absolute number of dendritic cells in the mLN. Results are from 3 experiments with 11 to 13 mice/group. (C) Percentage of dendritic cells in the mLN that coexpressed CD103 and/or CD11b. Data are derived from 2 experiments with 8 to 9 mice/group. (D-E) Representative flow plots and quantification of the expression of host peptide-donor MHC-II complex (YAe) on donor-derived DCs (CD45.2+ TCRβ−, CD64− CD11c+ MHC-II+). Data are from 3 experiments (n = 11-14/group). (F-J) Lethally irradiated Balb/c mice were transplanted with B6 Rag-1−/− BM (5 × 106) together with 1 × 106 purified CD4+ T cells from WT or GM-CSF−/− mice. Control mice received H2-Ab1−/− (MHC-II-deficient) BM (5 × 106) and 1 × 106 purified CD4+ from WT B6 donors. Sort-purified CD45.1+ CD4+ Vα2+ Vβ6+ cells (1 × 106) from TEa mice were adoptively transferred to mice 12 days posttransplantation. (F) Experimental design. (G) Representative dot plot gated on live, singlet cells from the colons of mice receiving H2-Ab1−/− BM and WT CD4+ T cells or Rag-1−/− BM and either WT or GM-CSF−/− CD4+ T cells 7 days posttransfer. (H) The frequency (expressed as a percentage of total live colon lamina propria cells) and absolute number of CD45.1+ CD4+ TEa cells in the colon 7 days posttransfer. Results are from 3 experiments (n = 9-11 per group). (I) Representative flow plots gated on CD4+ CD45.1+ (ie, TEa) cells depicting GM-CSF production. (J) The percentage of CD45.1+ CD4+ TEa cells in the colon of mice that expressed IFN-γ, GM-CSF, or TNF-α 7 days posttransfer. Results are from 2 experiments (n = 9/group). *P < .05; **P < .01; ***P < .001.

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