Figure 2
Figure 2. Donor T cells deficient for PKCα and θ produce reduced inflammatory cytokines in vivo. Lethally irradiated (800 cGy) BALB/c mice were transplanted with 5 × 106 genotype-matched TCD-BM cells alone (n = 2 for each genotype) or in addition to 1 × 106 T cells (CD4+, CD8+, and CD25–) from littermate WT C57BL/6 or PKCα−/−, PKCθ−/−, or PKCα−/−/θ−/− mice (n = 4 recipients per group per experiment). Two weeks posttransplant, recipient spleens (A) and livers (B) were harvested, and organ cell counts determined and stained for H2Kb, CD4, CD8, IFNγ, TNFα, and IL-4/5 analysis by flow cytometry. Absolute cell numbers depicted were calculated from whole spleen and liver counts at necropsy with flow cytometric percentages. Serum collected at necropsy was subjected to cytokine bead analysis (C) to quantify serum cytokine concentrations of IFN, TNF, IL-2, IL-6, IL-10, and IL-17. Cytokines that are not graphically represented were undetectable in serum. Averages for all values are represented with error bars indicating SEM from 1 representative experiment out of 3 separate experiments. *P < .05; **P < .01; ***P < .001 (compared with WT [brackets indicate statistical significance for comparisons between PKCα−/−/θ−/− and PKCθ−/− treatment groups]).

Donor T cells deficient for PKCα and θ produce reduced inflammatory cytokines in vivo. Lethally irradiated (800 cGy) BALB/c mice were transplanted with 5 × 106 genotype-matched TCD-BM cells alone (n = 2 for each genotype) or in addition to 1 × 106 T cells (CD4+, CD8+, and CD25) from littermate WT C57BL/6 or PKCα−/−, PKCθ−/−, or PKCα−/−−/− mice (n = 4 recipients per group per experiment). Two weeks posttransplant, recipient spleens (A) and livers (B) were harvested, and organ cell counts determined and stained for H2Kb, CD4, CD8, IFNγ, TNFα, and IL-4/5 analysis by flow cytometry. Absolute cell numbers depicted were calculated from whole spleen and liver counts at necropsy with flow cytometric percentages. Serum collected at necropsy was subjected to cytokine bead analysis (C) to quantify serum cytokine concentrations of IFN, TNF, IL-2, IL-6, IL-10, and IL-17. Cytokines that are not graphically represented were undetectable in serum. Averages for all values are represented with error bars indicating SEM from 1 representative experiment out of 3 separate experiments. *P < .05; **P < .01; ***P < .001 (compared with WT [brackets indicate statistical significance for comparisons between PKCα−/−−/− and PKCθ−/− treatment groups]).

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