[In vitro induction of allo-T lymphocytes proliferation by myeloid dendritic cells in patients with severe aplastic anemia.].
摘要
OBJECTIVE: To investigate the function of myeloid dendritic cells (mDCs) from severe aplastic anemia (SAA) patients in stimulating allogeneic T lymphocytes proliferation in vitro and then explore the immunopathogenesis of SAA. METHODS: Twenty-five SAA patients (15 untreated and 10 recovered after immunosuppressive therapy) and 12 normal controls were enrolled in this study. Their mature mDCs were induced from their bone marrow monocytes with recombined human interleukin-4 (rhIL-4), recombined human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and recombined human tumor necrosis factor (rhTNF) in vitro. Then mDCs were co-cultured with allogeneic lymphocytes (mixture lymphocyte reaction, MLR) at a ratio of 1:100 or 1:50. The growth rate of lymphocyte was measured with methyl thiazolyl tetrazolium (MTT) colorimetry. The concentrations of interleukin (IL)-12 and interferon gamma (IFNgamma) in MLR supernatant were measured with ELISA. The correlation between the growth rate and the concentration of IL-12 or IFNgamma was analyzed. RESULTS: When mDCs and lymphocytes were co-cultured at the ratio of 1:100, the growth rates of lymphocytes stimulated with mDCs from untreated, recovered SAA patients and controls were (219.8 +/- 94.0)%, (159.1 +/- 66.0)% and (160.1 +/- 91.9)% respectively. The concentrations of IL-12 in MLR supernatant were (8.2 +/- 3.6) ng/L, (6.5 +/- 2.8) ng/L and (6.1 +/- 2.6) ng/L and the concentrations of IFNgamma were (21.8 +/- 8.7) ng/L, (25.5 +/- 9.1) ng/L and (22.6 +/- 7.8) ng/L respectively. All of them had no statistical differences among the three groups (P > 0.05). When mDCs and lymphocytes were co-cultured at the ratio of 1:50, the growth rate of lymphocytes stimulated with mDCs from untreated patients was (322.1 +/- 171.1)%, which was higher than that of recovered patients [(180.9 +/- 79.1)% and controls (192.3 +/- 91.9)%]. The concentrations of IL-12 in MLR supernatant in the three groups were (12.6 +/- 4.4) ng/L, (9.4 +/- 3.3) ng