Article Figures & Data
Figures
- FIGURE 1.
Pulmonary metastasis in mice various days after inoculation of 2 × 105 B16F10 (BF10 mice) or B16F1 melanoma cells (BF1 mice) per mouse. The metastatic colonies were counted on days 3, 6, 9, 12, and 14 after the tumor inoculation in BF10 mice (•) or BF1 mice (○). Values are the average number of pulmonary colonies obtained from five mice. Results are the mean ± SE.
- FIGURE 2.
Production of IL-4 by CD4+ T cells from BF1 mice or BF10 mice. For induction of IL-4, CD4+ T cells (2 × 106 cells/ml) prepared from spleens of BF1 mice or BF10 mice on days 3, 7, and 14 after tumor inoculation were stimulated in vitro with anti-CD3 mAb. As a control, CD4+ T cells from normal mice (naive CD4+ T cells) were stimulated with mAb. Culture supernatants, harvested 48 h after stimulation, were assayed for IL-4 by ELISA. Results are the mean ± SD of triplicate determinations.
- FIGURE 3.
Pulmonary metastasis in BF1 mice inoculated with BF10-Th2 cells. Three days after inoculation of 2 × 105 B16F1 melanoma cells/mouse, mice (BF1 mice) received various numbers of splenic CD4+ T cells obtained from BF1 (○) or BF10 (•) mice at 14 days after their tumor implantation. As a control, BF1 mice were inoculated with the same number of CD4+ T cells from normal mice (naive CD4+ T cells) (▴). Fourteen days after inoculation with tumor cells, the number of metastatic colonies in lungs of these mice (five mice each) was examined, as described in text. Results are the mean ± SE.
- FIGURE 4.
Amount of pulmonary metastases in BF1 mice treated with IL-4. Mice (BF1 mice), at 3 days after i.v. inoculation with 2 × 105 B16F1 melanoma cells/mouse, were treated i.p. with various doses of IL-4 once a day for 7 days beginning just after tumor inoculation (10 mice each) (•). As a control, BF1 mice were treated with saline (○). The numbers of metastatic colonies in lungs of these mice were counted at 14 days after the tumor implantation, as described in text. The results expressed are the mean ± SE.
Tables
Cell Preparation Cytokinesa IL-2 (U/ml) IFN-γ (pg/ml) IL-4 (pg/ml) IL-10 (pg/ml) Naive CD4+ T cellsb 220 ± 37 560 ± 75 <50 <30 CD4+ T cells from BF1-micec 229 ± 34 495 ± 91 <50 <30 BF10-Th2 cellsd <10 <30 2670 ± 150 1500 ± 420 -
a Cell preparations were stimulated with anti-CD3 mAb (2.5 μg/ml) for 48 h to induce cytokine production. The culture fluids were assayed for cytokines by ELISA. The results expressed are the mean ± SD of triplicate determinations.
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b Naive CD4+ T cells were obtained from spleens of normal mice.
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c CD4+ T cells were prepared from mice 14 days after inoculation of 2 × 105 cells/mouse (i.v.) of B16F1 melanoma cells.
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d BF10-Th2 cells were CD4+ T cells from spleens of mice at 14 days after inoculation of B16F10 melanoma cells (2 × 105 cells/mouse).
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Cellsa IL-4 (pg/ml)b Decrease (%) Naive CD4+ T cells <50 BF10-Th2 cells (Th2 cells) 2515 ± 422 0 Th2 cells depleted of CD11b+ cells 120 ± 50c 95 Th2 cells depleted of CD28+ cells 87 ± 31c 97 Th2 cells depleted of TCR-αβ+ cells 58 ± 15c 98 Th2 cells depleted of TCR-γδ+ cells 2380 ± 360 5 -
a BF10-Th2 cells, CD4+ T cells prepared from mice at 14 days after inoculation of B16F10 melanoma cells (2 × 105 cells/mouse), were treated with various mAbs followed by complement, as described in text.
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b All cell preparations were stimulated with anti-CD3 mAb (2.5 μg/ml) for 48 h to induce IL-4 production. Culture fluids were assayed for IL-4 by ELISA. The results expressed are the mean ± SD of triplicate determinations.
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c Student’s t test, p < 0.001 vs original Th2 cells.
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- Table III.
Effect of anti-IL-4 mAb on pulmonary metastasis in BF10 mice or BF1 mice inoculated with BF10-Th2 cells
Mice No. of Pulmonary Metastasesa Reduction (%) BF10 mice receivedb Saline (control) 244 ± 22 0 Rat Ig 226 ± 37 7 Anti-IL-4 mAb 37 ± 16c 85 BF1 mice receivedd Saline 19 ± 5 BF10-Th2 cells (control) 347 ± 58 0 BF10-Th2 cells and rat Ig 326 ± 69 6 BF10-Th2 cells and anti-IL-4 mAb 60 ± 33c 83 -
a An average number of metastatic colonies per lungs of mice (10 mice each). Results are mean ± SE.
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b At 4 and 6 days after inoculation with B16F10 melanoma cells (2 × 105 cells/mouse), mice (BF10 mice) were treated i.p. with a 250-μg/kg dose of anti-IL-4 mAb. As controls, the same mice were treated with saline or rat Ig (250 μg/kg).
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c p < 0.01 vs an appropriate control.
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d Three days after the implantation of B16F1 melanoma cells (2 × 105 cells/mouse), mice (BF1 mice) were treated with saline or inoculated with 5 × 106 cells/mouse of BF10-Th2 cells. BF10-Th2 cells were CD4+ T cells prepared from spleens of mice at 14 days after the inoculation of 2 × 105 cells/mouse of B16F10 melanoma cells. BF1 mice inoculated with BF10-Th2 cells were treated i.p. with a 250-μg/kg dose of anti-IL-4 mAb or rat Ig 4 and 6 days after the tumor implantation.
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