American Society of Hematology

Figure 6

$Figure 6. VEGF inhibits the ability of DCs to stimulate allogeneic T-cell proliferation via both of VEGFR-1 and VEGFR-2, and blockade of VEGFR-2 is sufficient to restore DC function. The 8- to 10-week-old Balb/c mice were given PBS or VEGF receptor agonists for 28 days as described in Figure 1. (A) Expression of CD11c and CD86 in CD3e−B220− splenocytes was analyzed by FACS. The representative FACS plots were shown. The numbers represent the means of the percentage of indicated cell fractions. The first plot is the isotype control. (B) The ability of DCs to stimulate allogeneic T-cell proliferation. DCs were isolated from splenocytes with CD11c Microbeads. DCs (105) were incubated with 4 × 105 T cells (C57BL/6) for 4 days and 3H-thymidine uptake was analyzed in triplicate. (C) Balb/c mice were given PBS treated with rat IgG, or rhVEGF-treated rat IgG, anti-R1, or anti-R2 as described in Figure 1C. Expression of CD11c and CD86 in CD3e−B220− splenocytes was analyzed by FACS. The representative FACS plots were shown. The numbers represent the means of the percentage of indicated cell fractions. The first plot is the isotype control. (D) The ability of DCs to stimulate allogeneic T-cell proliferation. DCs were isolated from splenocytes with CD11c Microbeads. DCs (105) were incubated with 4 × 105 T cells (C57BL/6) for 4 days and 3H-thymidine uptake was analyzed in triplicate. The data (mean ± SEM, n = 3) repeat 2 times.$

VEGF inhibits the ability of DCs to stimulate allogeneic T-cell proliferation via both of VEGFR-1 and VEGFR-2, and blockade of VEGFR-2 is sufficient to restore DC function. The 8- to 10-week-old Balb/c mice were given PBS or VEGF receptor agonists for 28 days as described in Figure 1. (A) Expression of CD11c and CD86 in CD3e⁻B220⁻ splenocytes was analyzed by FACS. The representative FACS plots were shown. The numbers represent the means of the percentage of indicated cell fractions. The first plot is the isotype control. (B) The ability of DCs to stimulate allogeneic T-cell proliferation. DCs were isolated from splenocytes with CD11c Microbeads. DCs (10⁵) were incubated with 4 × 10⁵ T cells (C57BL/6) for 4 days and ³H-thymidine uptake was analyzed in triplicate. (C) Balb/c mice were given PBS treated with rat IgG, or rhVEGF-treated rat IgG, anti-R1, or anti-R2 as described in Figure 1C. Expression of CD11c and CD86 in CD3e⁻B220⁻ splenocytes was analyzed by FACS. The representative FACS plots were shown. The numbers represent the means of the percentage of indicated cell fractions. The first plot is the isotype control. (D) The ability of DCs to stimulate allogeneic T-cell proliferation. DCs were isolated from splenocytes with CD11c Microbeads. DCs (10⁵) were incubated with 4 × 10⁵ T cells (C57BL/6) for 4 days and ³H-thymidine uptake was analyzed in triplicate. The data (mean ± SEM, n = 3) repeat 2 times.

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