Article Figures & Data
Figures
- FIGURE 1.
CXCR4 expression of peripheral CD4+ T cells and cultured CD4+ T cells. CXCR4 expression was analyzed by FACS. The data shown are from CD4+ T cells stained with an isotype-matched control mAb (A), stained with anti-CXCR4 mAb (B), from CD4+ T cells cultured with medium for 6 h (C), or stimulated with anti-CD3 for 8 h (D) and stained with anti-CXCR4 mAb. Representative data from one of three independent experiments are shown.
- FIGURE 2.
SDF-1α up-regulates activation marker expression by anti-CD3-stimulated CD4+ T cells. Peripheral CD4+ T cells were cultured with medium for 6 h and then were stimulated in medium supplemented with 500 ng/ml SDF-1α for 2 h. Subsequently, the T cells were transferred to microtiter plates with or without coating with anti-CD3 mAb and were cultured for 8 h. Expression of CD69 (A), CD25 (B), and CD154 (C) was measured by FACS. Representative data from one of three independent experiments are shown. Presence of SDF-1α or anti-CD3 mAb is indicated.
- FIGURE 3.
SDF-1α up-regulates CD4+ T cell proliferation. Peripheral CD4+ T cells were cultured with medium for 6 h, and then the T cells were stimulated in medium supplemented with SDF-1α for 2 h. Subsequently, the T cells were transferred to microtiter plates with or without coating with anti-CD3 mAb and were cultured for 20 (A), 40 (B), and 60 h (C). The proliferation was measured by MTT assay and reported as OD units. Representative mean data from one of three independent experiments analyzed in triplicate are shown. Concentration of SDF-1α (ng/ml) and the presence of anti-CD3 mAb are indicated. ∗, p < 0.05; ∗∗, p < 0.01.
- FIGURE 4.
SDF-1α up-regulates cytokine production. Peripheral CD4+ T cells were cultured with medium for 6 h and then were stimulated in medium supplemented with SDF-1α for 2 h. Subsequently, the T cells were transferred to microtiter plates with or without coating with anti-CD3 mAb and were cultured for 8 h for IL-2 and IFN-γ production or for 12 h for IL-4 and IL-10 secretion. IL-2 (A), IFN-γ (B), IL-4 (C), and IL-10 (D) concentration in the culture supernatant was analyzed by ELISA. Representative mean data from one of three independent experiments analyzed in triplicate are shown. Concentration of SDF-1α (ng/ml) and the presence of anti-CD3 mAb are indicated. ∗, p < 0.05; ∗∗, p < 0.005.
- FIGURE 5.
Anti-CXCR4 mAb blocks enhancement of CD154 expression and IL-2 production by SDF-1α. Peripheral CD4+ T cells were cultured with medium for 5 h to up-regulate expression of CXCR4 and then were incubated with isotype-matched control mAb or anti-CXCR4 mAb for 1 h. Subsequently, they were stimulated in medium supplemented with 100 ng/ml SDF-1α for 2 h. The T cells were transferred to microtiter plates coated with anti-CD3 mAb and cultured for 8 h. A, Expression of CD154 was assessed by FACS. The data obtained from the cells incubated with isotype-matched control mAb (upper panels) and from the cells incubated with anti-CXCR4 mAb (lower panels) are depicted. Representative data from one of two independent experiments are shown. B, IL-2 concentration in the culture supernatant was analyzed by ELISA. Representative mean data from one of two independent experiments analyzed in triplicate are shown. Presence of SDF-1α, control mAb, or anti-CXCR4 mAb is indicated.
- FIGURE 6.
Comparison of costimulatory efficacy of SDF-1α and RANTES. Peripheral CD4+ T cells were cultured with medium for 6 h and then were stimulated in medium supplemented with 50 or 500 ng/ml SDF-1α or RANTES for 2 h. Subsequently, the T cells were transferred to microtiter plates coated with anti-CD3 mAb. After an 8-h incubation, expression of CD154 was assessed by FACS (A). The data obtained from the cells incubated without chemokines (left panel), from the cells costimulated with SDF-1α (center panel), and from the cells costimulated with RANTES (right panel) are depicted. Representative data from one of two independent experiments are shown. After 60 h of culture, proliferation was measured by MTT assay and reported as OD units (B). Representative mean data from one of two independent experiments analyzed in triplicate are shown. Concentrations of SDF-1α and RANTES are indicated. ∗, p < 0.01.
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